Effects of c-FLIPL Knockdown in Cervical Uterine Carcinoma Cell Lines
Effects of c-FLIPL Knockdown in Cervical Uterine Carcinoma Cell Lines
Authors
Ili, Carmen G.
Brebi, Priscilla
Garcia, Patricia
Leal, Pamela
Lopez, Jaime
Tapia, Oscar
Letelier, Pablo
Weber, Helga
Castillo, Jonathan
Roa, Juan C.
Brebi, Priscilla
Garcia, Patricia
Leal, Pamela
Lopez, Jaime
Tapia, Oscar
Letelier, Pablo
Weber, Helga
Castillo, Jonathan
Roa, Juan C.
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Abstract
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.