Skip to main content

Expressional variations of Kaiso: an association with pathological characteristics and field cancerization of OSCC

Abstract

Background

A group of genetically altered cells that have not transformed into a clinical or histologically identifiable state of malignancy but contains a higher risk of transforming into one is known as the field of cancerization. Numerous molecules are being investigated for their significance in the development of this phenomenon. One such protein of this family is Kaiso also known as ZBTB33 (Zinc Finger and BTB Domain containing 33). This protein belongs to the POZ-ZF family of transcription factors and may have functional tasks similar to its other siblings such as the growth and development of vertebrates and the pathogenesis of neoplastic diseases. Nevertheless, its role in the pathogenesis, progression, epithelial mesenchyal transition and field cancerization in case of oral cancer still needs exploration. Hence, this study was designed to explore the expressional differences between the mucosa of controls and those diagnosed with oral squamous cell carcinoma (OSCC).

Methods

Soft tissue samples were obtained from the main tumor, tumor periphery and opposite buccal mucosa of 50 oral cancer patients, whereas normal mucosa was taken from 50 volunteers undergoing elective tooth removal. The acquired samples were subjected to Immunohistochemical exploration for expression of Kaiso and E-Cadherin. The expression was measured using Image-J IHC profiler and summed as Optical density. The Optical density values were then subjected to statistical analysis.

Results

Results revealed a significant differential expression of Kaiso between the mucosal tissues taken from oral cancer patients and controls (p-value: < 0.0001), showing almost 50% down-regulation of Kaiso in all three tissue samples taken from oral cancer patients as compared to normal mucosa.

Conclusion

Kaiso has a significant difference of expression in the mucosa of oral cancer patients as compared to the mucosa of normal patients, making it a probable contributor to disease pathogenesis and field cancerization.

Peer Review reports

Background

Neoplasms arise due to compound genetic and epigenetic aberrations that consequentially transmute cells of a particular organ initiating an advanced invasive disease [1]. It is hypothesized that these genetic and epigenetic aberrations are not localized to a specific group of cells, rather they involve all the daughter cells residing in a particular field or surrounding tissues of the tumor [2]. Although not currently transformed completely in invasive disease, these resident cells harbour enough genetic anomalies which may transform them into malignant tumors at any given time in the future [2]. This populace of daughter cells residing in an organ, which has genetic aberrations but does not display morphological transformation consistent with malignant neoplasm, corroborates with the concept of field cancerization. This was initially presented in 1953 by Slaughter et al [3]. Scientific studies are being conducted to pinpoint molecular signatures which may help identify these genetically anomalous cell populations which have not adapted to the pathologic morphology of invasive carcinoma. When identified, these molecular markers may provide remarkable utility in terms of screening, diagnostics and targeted therapeutics. Some of the molecular markers found associated with oral field cancerization include cytokeratins 7, 8, 13, 16, and 19, [4] type 2 chain ABH antigen, [5] cyclin D1, [6, 7] EGFR, [8,9,10] TGF-α, [10] Ki-67, [11] BCL-2, [12] vascular markers (VwF, CD31, αVβ3, α-SMA), [13] and p53 [14,15,16,17,18,19]. One of the proteins of interest in this regard is Kaiso. Kaiso is a member of the broad-complex, tram track and bric-a-brac/poxvirus and zinc finger (BTB/POZ) family with a subfamily of zinc finger proteins (POZ-ZF) [20]. Transcription factors belonging to this family have been known to play a part in the growth and developmental aspects of vertebrates which indicates that Kaiso might have comparable functionality [21]. Kaiso’s functional preferences are described to change in context to its interactions with different proteins [22]. For instance, it has been demonstrated that Kaiso activates BCL-2 a protein that inhibits apoptotic death of the cell while deactivating the two pro-apoptotic proteins BAX and BIK hence leading to apoptotic diffidence [23]. Another example of context-specific functionality of Kaiso is demonstrated by its binding with wild type and mutated p53 where it promotes apoptosis when bonded with the former and suppresses programmed cell death when bonded with later, respectively [24, 25]. Considering the examples given previously Kaiso may be a vital part of pathogenic pathways leading to the development of neoplastic diseases. To date, ample scientific literature has verified Kaiso’s involvement in different types of cancers such as breast CA, [24, 26,27,28,29] Lung (NSCLC), [30] prostate CA, [31, 32] and pancreatic cancer (PDAC) [33]. It is also advocated that translocation of cytoplasmic fragment of E-cad into the nucleus modulates the activity of Kaiso, which exerts suppressive effects on the promoter region of target genes that are still mostly unidentified [34]. Having said that, depicting Kaiso as a tumor suppressor or tumor promoter with certainty. Furthermore, the scientific literature is also deficient regarding Kaiso’s functional role in the pathogenesis of oral mucosal cancer and devising Epithelial Mesenchymal Transition (EMT) has been a challenging task. It is yet to be explored whether the exposure of oral mucosa to known carcinogens such as tobacco, betel quid, betel nuts and other combination products has any effect on expressional values of Kaiso or not. Consequently, this study was directed at exploring expressional changes of Kaiso in mucosal specimens taken from the main tumor, the periphery of the tumor, and opposing mucosa of patients diagnosed with oral squamous cell carcinoma compared with subjects who were disease-free and were not exposed to any of the chemical carcinogens associated with the disease.

Methodology

Study design and study setting

The study design was analytical cross-sectional. Total soft tissue specimens from 50 biopsies of proven oral squamous cell carcinoma (OSCC) patients and 50 control tissues were obtained from patients who were attending the Department of Oral and Maxillofacial Surgery at Dow International Dental College, Dow University of Health Sciences, Karachi, Pakistan. The samples were collected during patients’ therapeutic surgery from OSCC cases and during elective tooth removal from controls, respectively. Cognizant patient consent was obtained from all study participants and endorsement was taken from the Institutional Review Board of Dow University (IRB-1319/DUHS/Approval/2019). Patients’ biographic information, medical history, extent and characteristics of the disease were documented in pre-designed proformas. Small tissue samples were taken from the Tumor (Labeled T), the periphery of the tumor (Labelled P), and the opposing mucosa (Labeled O), of OSCC patients who matched the inclusion and exclusion criteria (Table 1). From controls who matched the inclusion and exclusion criteria (Table 1), a small tissue specimen was collected during elective surgical removal of wisdom teeth, labelled as C. All samples were immediately placed in 10% buffered Neutral formalin.

Table 1 Inclusion and exclusion criteria for cases and controls

Sample size

The power of the test was calculated to justify the sample size of tissues per group using PASS version 15 software (NCSS, Kaysville, Utah, USA), based on a one-way analysis of variance test with 95% confidence of interval, an effect size of 0.854374 with 50 tissues in each of four groups computed using results from the expression of kaiso in mucosal samples taken from Tumor, Tumor’s periphery and Opposing cheek of OSCC patient. It was found to be more than 99%. The same power of the test was found using results from relationship between Kaiso and E-Cadherin with 95% confidence of interval, an effect size of 0.5537 with 50 tissues in each of four groups.

Tissue processing and immunohistochemical staining

The tissues obtained were embedded in paraffin to form blocks. Ultra-thin consecutive sections of 4 µm were incised from every single block. To scrutinize histopathological physiognomies, a section from each sample was stained with Hematoxylin & Eosin stains. After analyzing histologic characteristics of the tissues, we probed them for expression of Kaiso and E-Cadherin using Immunohistochemical staining. Briefly, the tissue sections were deparaffinized by xylene, after which tissue hydration was done, followed by heating in the microwave with citrate buffer (pH = 6) for retrieval of antigens. The tissues were then subjected to incubation in hydrogen peroxide for 20 min to mollify endogenous peroxidase interactions. Subsequently, tissue specimens were exposed to 15 bovine serum albumin for an hour to terminate the non-specific binding of antibodies. Next, tissues were incubated with polyclonal rabbit anti-kaiso (Thermo Fischer Scientific; PA5-81,890) and monoclonal mouse anti-E-cadherin (Thermo Fischer Scientific; 4A2c7) antibodies which were prepared at a dilution of 1:50. Human gall bladder tissues were used as a positive control for anti-kaiso, whereas gastric cancer tissues were used as positive controls for anti-E-cadherin antibodies as described in the catalogue provided by the manufacturer. After incubation with the Kaiso and E-cadherin, antibody slides were washed using Phosphate Buffered Saline. Then, EnVision FLEX, High pH (Link) (DAKO, Agilent Tech.) was used for detecting primary antibodies. Finally, the tissues were incubated with diaminobenzidine (DAB) + chromogen followed by a hematoxylin stain. After completing the chemical treatment, tissue slides were mounted using DPX and coverslip.

Imaging and expressional analysis

IHC slides from both cases and controls were photographed using Nikon Eclipse 80i at 40 × which are provided in the supplementary data upon reviewer’s request. To quantify the protein expression images were then analyzed using ImageJ IHC profiler software The software semi-quantifies the protein in the studied samples by assigning an automated score in a four tier system: high positive (HP), positive (P), low positive (LP) and negative (N). This is achieved by measuring pixel intensity that range from 0 to 255, where 0 represents the darkest shade and 255 represent the lightest shade of color as standard. The histogram profile zones are equally divided on the pixel color intensity bar. All intensities from 0 to 60 score for high positive zone, 61–120 for positive zone, 121 to 180 for low positive zone and 181 to 235 for negative zone. Values ranging from 235 to 255 represent fatty tissues and are excluded from the score determination zones. The four values obtained from ImageJ were then converted to optical density score, via formula given by S.Jafari et al. [34]:

$$(4\mathrm{HP}+3\mathrm{P}+2\mathrm{LP}+1\mathrm{N}) / 100$$

The optical density values were then used for statistical analysis.

Statistical analysis

Data normality was analyzed using the Kolmogorov–Smirnov test. Differential expression of the Kaiso and E-cadherin protein among the cases and controls, different tumor sizes and tumor grades was analyzed using one-way ANOVA. Pearson’s r test was done to find the correlation between the Expression of Kaiso and all the histological features recorded. The statistical analysis was performed employing Graph Pad Prism software and a p-value of less than 0.05 was considered significant. Also, all the results were further analysed as (positive + low positive) values VS high positive values as per advice of the reviewer but no significant differences were present than the cumulative results obtained as optical density. Hence these have not been included in the manuscript, although the file is added to the supplementary material.

Results

Out of total 50 cases 41 were obtained from males, whereas 9 were obtained from the females. The mean age of OSCC case group was 50.8 ± 11 years. Further stratification of the cases was done according to Broders grading System Table 2, and according to tumor size Table 3.

Table 2 Distribution of cases according to broders grading system [35]
Table 3 Distribution of cases according to tumor size

Differential analysis

Differential expression of Kaiso was assessed between tissue specimens taken from controls and tissue specimen taken from OSCC patients to determine expressional variations between the two and to assess if there is any kind of evidence present regarding field cancerization. Furthermore, expressional variations were assessed in different grades and sizes of tumors to determine Kaiso’s role in disease progression.

KAISO Expression in controls and tumor, periphery and opposing buccal mucosa of OSCC patients

When expression of Kaiso was compared among the two groups specimens taken from tumor (T), periphery (P), and opposite buccal mucosa (O); no significant differential expression was seen among the three groups in OSCC cases of Kaiso (p-value 0.1646), Fig. 1(a). But when all three values were analyzed against the expressional values of Control group a significant difference of expression was observed between tissues of controls and cases with a P-value of < 0.0001, Fig. 1(b). Mean expression values of Kaiso protein in all specimen are given in Table 4 showing a drastic decrease in kaiso expression in all tissues obtained from OSCC group.

Fig. 1
figure 1

Differential Expression of Kaiso in OSCC (a) and Controls against OSCC (b). The graph in figure a demonstrate the difference of expression in Kaiso among tissue specimens taken from OSCC cases; whereas figure b demonstrates differential expression between the Controls and OSCC tissue specimens. All values plotted here are in the form of optical density (OD). The large central line represents mean, whereas small horizontal lines above and below represent the standard deviation

Table 4 Mean expression of kaiso in cases and controls

E-Cadherin expression in controls and tumor, periphery and opposing buccal mucosa of OSCC patients

When expression of E-Cadherin was compared among the two groups specimens taken from tumor (T), periphery (P), and opposite buccal mucosa (O); no significant differential expression of E-Cadherin was seen among the three groups in OSCC cases (p-value 0.1566), Fig. 2(a). But when all three values were analyzed against the expressional values of Control group a significant difference of expression was observed between tissues of controls and cases with a P-value of < 0.0001, Fig. 2(b). Mean expression values of E-Cadherin protein in all specimen are given in Table 5 showing a drastic decrease in E-Cadherin expression in all tissues obtained from OSCC group as compared to Controls (P-value: < 0.0001).

Fig. 2
figure 2

Differential Expression of E-Cadherin in OSCC (a) and Controls against OSCC (b). The graph in figure a demonstrate the difference of expression in E-Cadherin among tissue specimens taken from OSCC cases; whereas figure b demonstrates differential expression between the Controls and OSCC tissue specimens. All values plotted here are in the form of optical density (OD). The large central line represents mean, whereas small horizontal lines above and below represent the standard deviation

Table 5 Mean expression of e-cadherin in cases and controls

KAISO expression in different grades and sizes of OSCC

On comparison between the different grades of tumor, no significant alteration could be observed among the mean expression values of Kaiso among different grades of OSCC, as confirmed by p-value 0.4042, Fig. 3(a). Similarly, no significant mean expressional variation could be found in different tumor (T) sizes (p3-value: 0.3762), Fig. 3(b). Mean expression values of Kaiso in different grades and T sizes are given in Table 6.

Fig. 3
figure 3

Differential Expression of Kaiso in Different Tumor Grades (a) and Tumor Sizes (b). The graph in figure a demonstrate the difference of expression in Kaiso among different Tumor Grades; whereas figure b demonstrates differential expression of Kaiso among different tumor sizes. All values plotted here are in the form of optical density (OD). The large central line represents mean, whereas small horizontal lines above and below represent the standard deviation

Table 6 Mean expression of kaiso in different tumor grades and sizes

Correlation analysis

Expression of kaiso was correlated with various histological features such as histological features defined in bryne’s TIF scoring system [36] and also total malignancy score (Table 7); Fig. 4, number of positive lymph nodes Fig. 5(a), tumor depth Fig. 5(b), and tumor budding scores [37] (Table 8), Fig. 5(c). No significant positive or negative correlation could be found or any of the physio-pathological characteristics as depicted by the straight correlation lines of the graphs and p-values mentioned in the respective figures. Similarly, Expression of Kaiso was correlated with E-Cadherin, Fig. 6. The only significant correlation could be observed between the two is in the region of tumor periphery. No other significant correlations could be observed.

Table 7 Bryne’s Tumor Invasive Front (TIF) Grading System [36]
Fig. 4
figure 4

Correlation between Expression of Kaiso and Histological Feature Scores from Bryne’s TIF Grading System. The graphs here demonstrate the correlation between expression of Kaiso in OSCC compiled as Optical Density and scores given to each histological feature according to Bryne’s scoring system. Each histological feature is correlated separately followed by the correlation with the total malignancy score

Fig. 5
figure 5

Correlation between Expression of Kaiso with Tumor Positive Lymphnodes (a), Tumor Depth (b) and Tumor Budding Score (c). The graphs here demonstrate expression of Kaiso in OSCC compiled as Optical Density, correlated with number of tumor positive lymphnodes, tumor depth measured in centimeters and tumor budding score

Table 8 Tumor budding score [37]
Fig. 6
figure 6

Correlation between Expressions of Kaiso with E-Cadherin. The graphs here demonstrate expression of Kaiso correlated with expression of E-Cadherin in OSCC (T, P, O) and Controls

Discussion

The concept of field cancerization was first suggested by Slaughter et al. after extensive histological examination of 783 cases of oral cancer, where he observed multiple centers of abnormal epithelium beyond the boundaries of the clinically evident malignant tumor. He observed that there was evidence of multiple microscopic foci of abnormal epithelial cells which later enlarge and coalesce to form a clinical picture of invasive carcinoma [3]. Later on, the term “Field Cancerization” was coined for this phenomenon and extensive scientific investigations on the molecular level were started regarding this concept to determine subsequent genetic aberrations, which is the driving force toward the malignant transformation of cells. Several genes and their expressional abnormalities have been implied as markers for the identification of field cancerization. These include, tumor suppressor genes such as p53, [38,39,40] p16, [41] Cyclin D1, [42] p21, [40]; Proto-oncogenes like, Ras [42] and Erb1 [43]; growth factor receptors such as EGFR [44], VEGF [45], TGF-α [10], and CD34 [46]. In this study, we have made an effort to determine whether expressional dysregulation of Kaiso has any role in field cancerization.

Kaiso is a member of the BTB/POZ-ZF family of site-specific transcription factors. The functional activity of a transcription factor has been considered vacillating due to its inclination towards context-specific suppression and/or activation of various genes in different types of cells [22]. Various factors are anticipated as game changers in Kaiso’s functional specificity towards a tumor suppressor or activator role. One of the factors may be Kaiso’s ability to bind both methylated and non-methylated sequence-specific sites (bi-modal DNA binding) [47, 48]. the second factor that should be taken into account is its SUMOylation which may play a role in the bimodal functional abilities of the proteins [49]. Another factor that can be taken into account is whether Kaiso binds with wild-type p53 or mutated p53 which determines the final functional path chosen by the protein [24, 25]. In this study we compared the expression of Kaiso in the mucosa of subjects who were never exposed to chemical carcinogens known to predispose to oral cancer, with the expression of Kaiso in biopsy-proven tumors and seemingly normal mucosa taken from the tumor periphery and opposing cheek of the same patient. The idea was to determine what path this protein takes in case of oral squamous cell carcinoma in terms of expression. Astonishingly, the findings show an almost 50% decreased expression of Kaiso in the mucosa of OSCC patients in comparison to controls with a p-value of < 0.0001 (Fig. 1b). Also, it can be appreciated that pure tumor tissue, peripheral mucosa and opposing mucosa of OSCC cases, all show similar mean expressions of Kaiso (Table 4), indicating a decreased Kaiso expression in the entire oral cavity and not specifically the tumor. These findings highlight that Kaiso might have a vital role in defining the field of cancerization in cases of oral squamous cell carcinoma. Furthermore, levels of E-cadherin expressions in the same tissue specimens of OSCC was also found significantly down regulated (Fig. 2b, Table 5) as compared to controls but when the two proteins were correlated with each other, no significant correlation could be observed between the two except in the region of tumor periphery (Fig. 6), where a negative correlation could be observed between the two proteins. Also, a correlation analysis was done between the expressional variations of Kaiso with different tumor grades and tumor sizes which turned out to be insignificant (Fig. 3a and b). These outcome indicates that Kaiso’s expressional vicissitudes are more likely involved in the incidence of OSCC rather than its progression. The other inference that could be obtained from this study was that, although E-cadherin might be an independent marker of epithelial mesenchymal transition and field cancerization but it does not show any significant correlation with the expressional changes of kaiso in case if OSCC. Moreover, if E-cadherin affects kaiso in any way it is not in terms of expressional changes, rather it may affect Kaiso’s ability to alter the expression of target genes after binding with p120 as suggested by Van Roy et al [20]. Additionally, correlation analyses were done employing Kaiso expression in tumor tissues and the histological features of these specimens which are frequently used in terms of prognostic determinants of OSCC (Figs. 4 and 5), namely the number of positive lymph nodes, depth of tumor, number of tumor buds, degree of keratinization, number of mitotic figures, nuclear polymorphism, the pattern of invasion and extent of plasma-lymphocytic infiltrate. No significant correlation could be observed between the expression of Kaiso and histological features mentioned, which further strengthens our impression of Kaiso acting as a match stick to a haystack in the case of OSCC. A study by Cofre et al. demonstrated that diminution of Kaiso expression resulted in the augmented proliferation and reduced expression of differentiation markers; supporting our findings [50]. In contrast to this study there are several studies which have reported a higher expression of Kaiso associated with different types and pathological features of cancers. For instance, Pierre et al. reported a greater Kaiso expression in primary & metastatic tumor tissue specimens of colon, in comparison to normal [51]. Whereas, Jones et al. observed a higher Kaiso expression in malignant tumors of the prostate than benign prostatic hyperplasia. He also reported that higher expression of Kaiso had a correlation with higher tumor grade [31]. In another scientific paper, Jones et al. reported similar associations between high Kaiso expression in pancreatic ductal carcinomas and higher tumor grades and sizes [33]. Analogous findings are reported in the case of breast carcinoma in various studies [5, 7, 22, 23]. Differences which might be responsible for such contrasting results might include exposure to chemical carcinogens specific to the oral cavity such as betel nut and betel quid promoting a factor specific functioning of Kaiso.

What mechanism and/or factors are behind this striking decrease in Kaiso’s expression in the mucosa of oral cancer patients and whether Kaiso plays a role in devising epithelial mesenchymal transition in OSCC still needs to be explored, giving us a future direction for a much needed scientific research.

Conclusion

In conclusion, it can be safely assumed that Kaiso has a role in the genesis of Oral cancer indicated by its expressional dysregulation in OSCC patients. Kaiso may also serve as a marker for field cancerization in OSCC patients. These findings make Kaiso a suitable candidate for targeted therapy. Hence, it is recommended that other aspects of Kaiso’s functionality should be assessed to further elaborate its role in the pathogenesis of oral squamous cell carcinoma and field cancerization.

Limitations of the study

Like most studies, there were some limitations to this study as well. The measurement of protein was done by one method only. This may be performed with other methods of protein estimation to cross-check the results. No specimens were included from patients with precancerous conditions, the inclusion of which may help determine a threshold level of Kaiso that indicates the conversion of a premalignant state into a malignant state before the appearance of physiopathological characteristics. It is therefore required that the research is validated with improved sample size and multicenter study, with populations from different ethnicities, and with groups including oral premalignant states and lesions in future. Furthermore, survival analysis of the study could not be performed as the sample collection for this study was started in year 2019 (IRB ref no: IRB-1319/DUHS/Approval/2019) and continued till 2021, the patients are still being followed for survival data which will be published it in near future.

Availability of data and materials

The data collected and used for this research is the property of Dow University of Health Sciences, whereas the data used in this study is available upon request from the corresponding author.

Abbreviations

OSCC:

Oral Squamous Cell Carcinoma

IHC:

Immunohistochemistry

PBS:

Phosphate Buffered Saline

BNF:

Neutral Buffered Formalin

References

  1. Eljabo N, Nikolic N, Carkic J, Jelovac D, Lazarevic M, Tanic N, et al. Genetic and epigenetic alterations in the tumour, tumour margins, and normal buccal mucosa of patients with oral cancer. Int J Oral Maxillofac Surg. 2018;47(8):976–82.

    CAS  PubMed  Article  Google Scholar 

  2. Curtius K, Wright NA, Graham TA. An evolutionary perspective on field cancerization. Nat Rev Cancer. 2018;18(1):19–32.

    CAS  PubMed  Article  Google Scholar 

  3. Slaughter DP, Southwick HW, Smejkal W. “Field cancerization” in oral stratified squamous epithelium. Clinical implications of multicentric origin. Cancer. 1953;6(5):963–8.

    CAS  PubMed  Article  Google Scholar 

  4. Bosch FX, Ouhayoun J-P, Bader BL, Collin C, Grund C, Lee I, et al. Extensive changes in cytokeratin expression patterns in pathologically affected human gingiva. Virchows Archiv B. 1989;58(1):59.

    CAS  Article  Google Scholar 

  5. Bongers V, Snow GB, De Vries N, Braakhuis BJ. Potential early markers of carcinogenesis in the mucosa of the head and neck using exfoliative cytology. J Pathol. 1996;178(3):284–9.

    CAS  PubMed  Article  Google Scholar 

  6. Bartkova J, Lukas J, Müller H, Strauss M, Gusterson B, Bartek J. Abnormal patterns of D-type cyclin expression and G1 regulation in human head and neck cancer. Can Res. 1995;55(4):949–56.

    CAS  Google Scholar 

  7. Bloching MM, Soulsby H, Naumann A, Aust W, Merkel D, Braun T, et al. Tumor risk assessment by means of immunocytochemical detection of early pre-malignant changes in buccal smears. Oncol Rep. 2008;19(6):1373–9.

    PubMed  Google Scholar 

  8. Shin DM, Ro JY, Hong WK, Hittelman WN. Dysregulation of epidermal growth factor receptor expression in premalignant lesions during head and neck tumorigenesis. Can Res. 1994;54(12):3153–9.

    CAS  Google Scholar 

  9. Van Oijen M, Rijksen G, Broek Ft, Slootweg PJ. Increased expression of epidermal growth factor receptor in normal epithelium adjacent to head and neck carcinomas independent of tobacco and alcohol abuse. Oral Dis. 1998;4(1):4–8.

    PubMed  Article  Google Scholar 

  10. Grandis JR, Tweardy DJ. Elevated levels of transforming growth factor α and epidermal growth factor receptor messenger RNA are early markers of carcinogenesis in head and neck cancer. Can Res. 1993;53(15):3579–84.

    CAS  Google Scholar 

  11. van Oijen MG, Gilsing MMA, Rijksen G, Hordijk GJ, Slootweg PJ. Increased number of proliferating cells in oral epithelium from smokers and ex-smokers. Oral Oncol. 1998;34(4):297–303.

    PubMed  Article  Google Scholar 

  12. Birchall MA, Schock E, Harmon BV, Gobe G. Apoptosis, mitosis, PCNA and bcl-2 in normal, leukoplakic and malignant epithelia of the human oral cavity: prospective, in vivo study. Oral Oncol. 1997;33(6):419–25.

    CAS  PubMed  Article  Google Scholar 

  13. El-Gazzar R, Macluskey M, Ogden GR. Evidence for a field change effect based on angiogenesis in the oral mucosa? A brief report. Oral Oncol. 2005;41(1):25–30.

    CAS  PubMed  Article  Google Scholar 

  14. Cruz IB, Snijders PJ, Meijer CJ, Braakhuis BJ, Snow GB, Walboomers JM, et al. p53 expression above the basal cell layer in oral mucosa is an early event of malignant transformation and has predictive value for developing oral squamous cell carcinoma. J Pathol. 1998;184(4):360–8.

    CAS  PubMed  Article  Google Scholar 

  15. Lavieille JP, Brambilla E, Riva-Lavieille C, Reyt E, Charachon R, Brambilla C. Immunohistochemical detection of p53 protein in preneoplastic lesions and squamous cell carcinoma of the head and neck. Acta Otolaryngol. 1995;115(2):334–9.

    CAS  PubMed  Article  Google Scholar 

  16. Nakanishi Y, Noguchi M, Mukai K, Matsuno Y, Hirohashi S, Saikawa M, et al. p53 expression in multicentric squamous cell carcinoma and surrounding squamous epithelium of the upper aerodigestive tract. Immunohistochemical analysis of 95 lesions. Cancer. 1995;75(7):1657–62.

    CAS  PubMed  Article  Google Scholar 

  17. Nees M, Homann N, Discher H, Andl T, Enders C, Herold-Mende C, et al. Expression of mutated p53 occurs in tumor-distant epithelia of head and neck cancer patients: a possible molecular basis for the development of multiple tumors. Can Res. 1993;53(18):4189–96.

    CAS  Google Scholar 

  18. van Oijen MG, van de Craats JG, Slootweg PJ. p53 overexpression in oral mucosa in relation to smoking. J Pathol. 1999;187(4):469–74.

    PubMed  Article  Google Scholar 

  19. Waridel F, Estreicher A, Bron L, Flaman JM, Fontolliet C, Monnier P, et al. Field cancerisation and polyclonal p53 mutation in the upper aero-digestive tract. Oncogene. 1997;14(2):163–9.

    CAS  PubMed  Article  Google Scholar 

  20. Van Roy FM, McCrea PD. A role for Kaiso–p120ctn complexes in cancer? Nat Rev Cancer. 2005;5(12):956–64.

    PubMed  Article  CAS  Google Scholar 

  21. Kelly KF, Daniel JM. POZ for effect–POZ-ZF transcription factors in cancer and development. Trends Cell Biol. 2006;16(11):578–87.

    CAS  PubMed  Article  Google Scholar 

  22. Pierre CC, Hercules SM, Yates C, Daniel JM. Dancing from bottoms up–roles of the POZ-ZF transcription factor Kaiso in cancer. Biochim Biophys Acta Rev Cancer. 2019;1871(1):64–74.

    CAS  PubMed  Article  Google Scholar 

  23. Xue X, Zhang J, Lan H, Xu Y, Wang H. Kaiso protects human umbilical vein endothelial cells against apoptosis by differentially regulating the expression of B-cell CLL/lymphoma 2 family members. Sci Rep. 2017;7(1):1–16.

    Article  CAS  Google Scholar 

  24. Bassey-Archibong BI, Rayner LG, Hercules SM, Aarts CW, Dvorkin-Gheva A, Bramson JL, et al. Kaiso depletion attenuates the growth and survival of triple negative breast cancer cells. Cell Death Dis. 2017;8(3):e2689.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  25. Koh D-I, Han D, Ryu H, Choi W-I, Jeon B-N, Kim M-K, et al. KAISO, a critical regulator of p53-mediated transcription of CDKN1A and apoptotic genes. Proc Natl Acad Sci. 2014;111(42):15078–83.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  26. Vermeulen JF, van de Ven RA, Ercan C, van der Groep P, van der Wall E, Bult P, et al. Nuclear Kaiso expression is associated with high grade and triple-negative invasive breast cancer. PLoS ONE. 2012;7(5):e37864.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  27. Jones J, Wang H, Karanam B, Theodore S, Dean-Colomb WM, Welch DR, et al. Nuclear localization of Kaiso as a prognostic biomarker and promotion of EMT in infiltrating ductal breast carcinomas. Am Soc Clin Oncol. 2014.

  28. Bassey-Archibong BI, Hercules SM, Rayner LG, Skeete DH, Connell SPS, Brain I, et al. Kaiso is highly expressed in TNBC tissues of women of African ancestry compared to Caucasian women. Cancer Causes Control. 2017;28(11):1295–304.

    PubMed  PubMed Central  Article  Google Scholar 

  29. Kwiecien JM, Bassey-Archibong BI, Dabrowski W, Rayner LG, Lucas AR, Daniel JM. Loss of Kaiso expression in breast cancer cells prevents intra-vascular invasion in the lung and secondary metastasis. PLoS ONE. 2017;12(9):e0183883.

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  30. Dai S-D, Wang Y, Miao Y, Zhao Y, Zhang Y, Jiang G-Y, et al. Cytoplasmic Kaiso is associated with poor prognosis in non-small cell lung cancer. BMC Cancer. 2009;9(1):1–13.

    Article  CAS  Google Scholar 

  31. Jones J, Wang H, Zhou J, Hardy S, Turner T, Austin D, et al. Nuclear Kaiso indicates aggressive prostate cancers and promotes migration and invasiveness of prostate cancer cells. Am J Pathol. 2012;181(5):1836–46.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  32. Wang H, Liu W, Black S, Turner O, Daniel JM, Dean-Colomb W, et al. Kaiso, a transcriptional repressor, promotes cell migration and invasion of prostate cancer cells through regulation of miR-31 expression. Oncotarget. 2016;7(5):5677.

    PubMed  Article  Google Scholar 

  33. Jones J, Mukherjee A, Karanam B, Davis M, Jaynes J, Reams RR, et al. African Americans with pancreatic ductal adenocarcinoma exhibit gender differences in Kaiso expression. Cancer Lett. 2016;380(2):513–22.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  34. SeyedJafari SM, Hunger R. IHC optical density score: a new practical method for quantitative immunohistochemistry image analysis. Appl Immunohistochem Mol Morphol. 2017;25(1):e12–3.

    Article  Google Scholar 

  35. BRODERS AC. The microscopic grading of cancer. The surgical clinics of north America. 1941;21:947–62.

  36. Bryne M, Koppang HS, Lilleng R, Kjærheim Å. Malignancy grading of the deep invasive margins of oral squamous cell carcinomas has high prognostic value. J Pathol. 1992;166(4):375–81.

    CAS  PubMed  Article  Google Scholar 

  37. Boxberg M, Jesinghaus M, Dorfner C, Mogler C, Drecoll E, Warth A, et al. Tumour budding activity and cell nest size determine patient outcome in oral squamous cell carcinoma: proposal for an adjusted grading system. Histopathology. 2017;70(7):1125–37.

    PubMed  Article  Google Scholar 

  38. van Houten VM, Tabor MP, van den Brekel MW, Alain Kummer J, Denkers F, Dijkstra J, et al. Mutated p53 as a molecular marker for the diagnosis of head and neck cancer. J Pathol. 2002;198(4):476–86.

    PubMed  Article  CAS  Google Scholar 

  39. El-Naggar AK, Lai S, Luna MA, Zhou X-D, Weber RS, Goepfert H, et al. Sequential p53 mutation analysis of pre-invasive and invasive head and neck squamous carcinoma. Int J Cancer. 1995;64(3):196–201.

    CAS  PubMed  Article  Google Scholar 

  40. Reda E, Reda N. Over expression of p53 and p21 in normal oral mucosa adjacent to resected squamous cell carcinomas. 2005.

    Google Scholar 

  41. Izzo JG, Papadimitrakopoulou VA, Li XQ, Ibarguen H, Lee JS, Ro JY, et al. Dysregulated cyclin D1 expression early in head and neck tumorigenesis: in vivo evidence for an association with subsequent gene amplification. Oncogene. 1998;17(18):2313–22.

    CAS  PubMed  Article  Google Scholar 

  42. Ortiz BH, Ailawadi M, Colitti C, Muto MG, Deavers M, Silva EG, et al. Second primary or recurrence? Comparative patterns of p53 and K-ras mutations suggest that serous borderline ovarian tumors and subsequent serous carcinomas are unrelated tumors. Can Res. 2001;61(19):7264–7.

    CAS  Google Scholar 

  43. Werkmeister R, Brandt B, Joos U. The erbB oncogenes as prognostic markers in oral squamous cell carcinomas. Am J Surg. 1996;172(6):681–3.

    CAS  PubMed  Article  Google Scholar 

  44. Szymańska K, Levi JE, Menezes A, Wünsch-Filho V, Eluf-Neto J, Koifman S, et al. TP53 and EGFR mutations in combination with lifestyle risk factors in tumours of the upper aerodigestive tract from South America. Carcinogenesis. 2010;31(6):1054–9.

    PubMed  Article  CAS  Google Scholar 

  45. Astekar M, Joshi A, Ramesh G, Metgud R. Expression of vascular endothelial growth factor and microvessel density in oral tumorigenesis. J Oral Maxillofac Pathol. 2012;16(1):22.

    PubMed  PubMed Central  Article  Google Scholar 

  46. López-Blanc SA, Collet AM, Gandolfo MS, Femopase F, Hernández SL, Tomasi VH, et al. Nucleolar organizer regions (AgNOR) and subepithelial vascularization as field cancerization markers in oral mucosa biopsies of alcoholic and smoking patients. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2009;108(5):747–53.

    PubMed  Article  Google Scholar 

  47. Daniel: The p120 ctn-binding partner Kaiso is a bi-modal... - Google Scholar [

  48. Buck-Koehntop BA, Stanfield RL, Ekiert DC, Martinez-Yamout MA, Dyson HJ, Wilson IA, et al. Molecular basis for recognition of methylated and specific DNA sequences by the zinc finger protein Kaiso. Proc Natl Acad Sci. 2012;109(38):15229–34.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  49. Zhenilo S, Deyev I, Litvinova E, Zhigalova N, Kaplun D, Sokolov A, et al. DeSUMOylation switches Kaiso from activator to repressor upon hyperosmotic stress. Cell Death Differ. 2018;25(11):1938–51.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  50. Cofre J, Menezes JR, Pizzatti L, Abdelhay E. Knock-down of Kaiso induces proliferation and blocks granulocytic differentiation in blast crisis of chronic myeloid leukemia. Cancer Cell Int. 2012;12(1):1–14.

    Article  CAS  Google Scholar 

  51. Pierre C. Kaiso: a novel mediator of intestinal inflammation and tumorigenesis [PhD Thesis]. 2015.

    Google Scholar 

Download references

Acknowledgements

We are highly indebted to all the research participants for their constant co-operation during the study. We are also extremely grateful to Dow Research Institute of Biotechnology and Bio-Sciences for providing us access to all the necessary equipment for this research.

Funding

This is a self-funded research project.

Author information

Authors and Affiliations

Authors

Contributions

SA: Contributed to conceptualizing the overall study, data collection, sample processing and drafting of the manuscript. SK: Supervised as well as helped in drafting and editing of the manuscript. MAQ: Co-Supervised the study in addition to drafting and editing the main manuscript. UB: Contributed and helped in the assessment of patients with Oral Squamous Cell Carcinoma. MA: Contributed to analyzing the results of the study. MNM: Co-Supervised the study and contributed to conceptualizing the overall study. The author(s) read and approved the final manuscript.

Corresponding authors

Correspondence to Saeed Khan or Muhammad Nouman Mughal.

Ethics declarations

Ethics approval and consent to participate

This study has been conducted after taking endorsement from the Institutional Review Board of Dow University of Health Sciences IRB No: IRB-1319/DUHS/Approval/2019. All techniques employed in the study were in agreement with the 1964 Helsinki Declaration and the standards approved by the institutional and/or national research committee. Informed consent was obtained in writing from all the research participants.

Consent for publication

The current study does not contain any images or videos related to any research participant. The consent for publication is obtained from all research participants included in the current study.

Competing interests

The authors declare that they have no conflict of interest.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Ahmed, S., Khan, S., Qureshi, M.A. et al. Expressional variations of Kaiso: an association with pathological characteristics and field cancerization of OSCC. BMC Cancer 22, 990 (2022). https://doi.org/10.1186/s12885-022-10014-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1186/s12885-022-10014-7

Keywords

  • Kaiso
  • E-Cadherin
  • Oral cancer
  • Field cancerization
  • Oral squamous cell carcinoma
  • Immunohistochemistry (IHC)