Relevance of cyclin D1b expression and CCND1polymorphism in the pathogenesis of multiple myeloma and mantle cell lymphoma

Table 2 Cyclin D1a and cyclin D1b mRNA expression in MCL and MM samples

Sample	ΔCta	ΔΔCta	Na	ΔCtb	ΔΔCtb	Nb
MCL
JeKo-1*	6.5	0	1	12.7	0	1
GRANTA-519	12.5	6.0	0.0156	19.0	6.3	0.0127
HBL-2	7.8	1.3	0.4061	14.5	1.8	0.2871
NCEB-1	10.9	4.4	0.0474	19.7	7.0	0.0078
Pt 1	10.4	3.9	0.0670	19.3	6.6	0.0103
Pt 2	11.3	4.8	0.0359	16.4	6.7	0.0096
Pt 3	10.1	5.6	0.0206	nc	nc	nc
Pt 4	12.9	6.4	0.0118	18.2	8.5	0.0028
Pt 5	9.2	2.7	0.1539	15.5	5.8	0.0179
Pt 6	8.8	2.3	0.2030	15.0	5.3	0.0254
Pt 7	11.4	4.9	0.0335	16.9	7.2	0.0068
Pt 8	9.1	2.6	0.1650	14.3	4.6	0.0412
Pt 9	12.2	5.7	0.0192	17.0	7.3	0.0063
MM
U266*	10.0	0	1	18.8	0	1
Karpas 620	13.9	3.9	0.0670	20.8	2.0	0.2500
RPMI 8226	17.1	7.1	0.0073	23.9	5.1	0.0291
NCI-H929	15.9	5.9	0.0167	23.7	4.9	0.0335
OPM-2	nc	nc	nc	nc	nc	nc
Pt 1	12.3	2.3	0.2030	18.5	-0.3	1.2311
Pt 2	18.2	8.2	0.0034	27.7	8.9	0.0021
Pt 3	15.0	5.0	0.0312	27.1	8.3	0.0032

The quantitation of cyclin D1a and cyclin D1b mRNA compared to 18S rRNA was done using the comparative threshold (Ct) method with the ABI PRISM 7000 SDS software. Triplicate experiments were done for each sample; for each sample the average Ct value for the internal standard was subtracted from the average Ct value for cyclin D1a or cyclin D1b to yield ΔCta and ΔCtb. ΔCt obtained from the calibrator cell lines (* JeKo-1 for MCL samples and U266 for MM samples) was subtracted from each ΔCta or ΔCtb to give the ΔΔCt. The relative amount of cyclin D1a and cyclin D1b compared to the calibrator was calculated by the formula N = 2^-ΔΔCt. No cyclin D1b was detected in MCL patient 3 (Ctb>50); no cyclin D1a or b was detected in OPM-2 MM cell line (Cta>50 and Ctb>50). nc, not calculated; Pt, patient.

ISSN: 1471-2407