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Figure 2 | BMC Cancer

Figure 2

From: The tumour-promoting receptor tyrosine kinase, EphB4, regulates expression of Integrin-β8 in prostate cancer cells

Figure 2

Knockdown ofITGB8results in reduced metastatic potential in prostate cancer cells. A) Quantitative real-time PCR was carried out to determine knockdown levels of siRNA against ITGB8. 100 nM negative non-silencing or ITGB8 targeting siRNA were transiently transfected into PC-3 cells. RNA was isolated 48 h after transfection, transcribed into cDNA and analyzed for gene expression. ITGB8 expression is reduced by approximately 60-70%. B) PC-3 cells were transiently transfected with 100 nM negative non-silencing (neg si) or ITGB8 targeting siRNA (β8 si) and 24 h later a scratch wound was applied using the IncuCyte (Essen Bioscience) system and migration was monitored for a further 24 h. Cell migration was reduced following knockdown of ITGB8. C) PC-3 cells were transiently transfected with 100 nM negative non-silencing (neg si) or ITGB8 targeting siRNA (β8 si) and subjected to a Matrigel transwell invasion assay. After 22 h of incubation, invaded cells were stained and counted. Cell invasion was reduced following knockdown of ITGB8. D) 22Rv1-VO (vector only) or 22Rv1–B4 (EphB4 over-expressing) cells were transiently transfected with 100 nM siRNA against ITGB8 (β8 si) or negative non-silencing siRNA (neg si). An invasion assay was carried out using the Matrigel invasion system and cells were allowed to invade for 22 h. Cells containing the ITGB8 siRNA showed significantly reduced ability to invade. E) 22Rv1-VO (vector only) or 22Rv1–B4 (EphB4 over-expressing) cells were transiently transfected with siRNA against ITGB8 (β8 si) or negative non-silencing siRNA (neg si) and subjected to an adhesion assay to vitronectin. No significant changes were seen. n = 3 * p < 0.01 vs VO negative; # p < 0.001 vs B4 negative.

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