RLIP76 silencing inhibits cell proliferation and invasion in melanoma cell line A375.

OBJECTIVE

RLIP76 is a stress-responsive membrane protein regulating multiple cellular signaling pathways and is overexpressed in a number of malignant tumor cells. The aim of this study is to test whether the RLIP76 plays a critical role in cellular growth, apoptosis, cell cycle distribution and chemoresistance in melanoma.

MATERIALS AND METHODS

In this study, human melanoma cell line A375 was stably transfected with an RLIP76-targeted shRNA-containing vector to investigate the role of RLIP76 in cellular function. Quantitative RT-PCR and Western blot revealed significant suppression of RLIP76 in A375 cells after transfection with shRNA-containing vector. Cell proliferation was determined by MTT assay. Wound scratch ad transwell assays were used for cell migration and invasion.

RESULTS

The results showed that shRNA decreased the expression of RLIP76 in human melanoma cell line A375, and the knockdown of RLIP76 gene significantly inhibited A375 cell growth, resulted in G1 phase arrest and apoptosis increase of A375. Moreover, by testing the cell migratory and invasive abilities by wound scratch and transwell assays, it determined that the RLIP76 also suppressed cell migration and invasion in A375 cells and P13K/Akt signalling CONCLUSIONS: Overall, this study suggests that RLIP76 is a potential therapeutic target against melanoma.