Effects of c-FLIPL knockdown in cervical uterine carcinoma cell lines.

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Carmen G. Ili; Priscilla Brebi; Patricia Garcia; Pamela Leal; Jaime Lopez; Oscar Tapia; Pablo Letelier; Helga Weber; Jonathan Castillo & Juan C. Roa

Summary

Overexpression of Short and Raji variants of Cellular FLICE-like inhibitory protein (c-FLIP) is capable of inhibiting apoptosis, while the function of the Long isoform depends of c-FLIPL concentration in cells. The aim of this study was to determine the effects of c-FLIPL knockdown in cervical cell lines. SiHa, C-4I and C-33A cervical cancer cell lines were analyzed. c-FLIPL level expression was determined by quantitative real-time PCR and western blotting. c-FLIPL was transiently downregulated by siRNA. The effects of knockdown of c-FLIPL on cell viability, proliferation and apoptosis were assessed by comparing with scrambled siRNA- transfected cells. SiHa and C-4I c-FLIPL knockdown cells showed increased viability compared with scrambled siRNA-transfected cells (P<0.05), while C-33A cells did not show significant differences. Ki-67 and PCNA immunocytochemistry was performed to evaluate proliferation on these cervical cancer cell lines. SiHa cells with c-FLIPL knockdown showed elevated expression of Ki-67 protein compared with their scrambled counterparts (P<0.0001), while C-33A c-FLIPL knockdown cells showed a significantly lower in PCNA expression (P<0.01) compared with control. All three c-FLIP-transfected cell lines showed a higher level of apoptosis compared with their scrambled controls. Our results suggest that c-FLIPL could have effects in proliferation and apoptosis in cervical cancer cell lines.

KEY WORDS: Apoptosis; Proliferation; Cervical cancer; siRNA; c-FLIPL.

How to cite this article

ILI, C. G.; BREBI, P.; GARCIA, P.; LEAL, P.; LOPEZ, J.; TAPIA, O.; LETELIER, P.; WEBER, H.; CASTILLO, J. & ROA, J. C. Effects of c-FLIPL knockdown in cervical uterine carcinoma cell lines. Int. J. Morphol., 33(2):638-646, 2015.