Dados do Resumo

Título

PROFILE OF HUMAN EPITHELIAL RECEPTORS IN OVARIAN CELL LINES

Introdução

Ovarian cancers have become the second most common malignancy in women after breast cases and mortality rates have remained relatively stable, with a survival rate around 30% over 5 years in the past 40 years. This work investigated the role of human epithelial receptors (HERs) for cell survival of ovarian tumor lines.

Objetivo

This work investigated the role of human epithelial receptors (HERs) for cell survival of ovarian tumor lines.

Métodos

Initially, seven distinct ovarian cancer lines (SKOV-3, OVCAR-8, IGROV-1, A-2780, OVCAR-3, PEO-4 E PEO-6) of high- and low-grade carcinomas were submitted to protein extraction, SDS-PAGE electrophoresis and immunoblotting for HER identification (HER 1, 2, 3 and/or 4). The quantification of the bands was performed using the ImageJ 1.44 software. Next, lentiviruses were used for silencing experiments with shRNA sequences cloned into the pLKO vector. To produce the viruses, HEK-293T cells transformed with the adenoviral genes E1a and E1b and containing the SV40 virus T antigen were used. Transfection of HEK-293T cells was performed with the jetPEITM reagent (PolyPlus Transfection). Then, target ovarian cells were infected and new medium with puromycin 3 µg/mL was added 72 h thereafter. Cell proliferation was determined by the MTT assay following 8, 72, 96 and 120 h of incubation. Gene silencing of HER3 was analyzed by western blotting.

Resultados

The profile of receptors showed expression of HER3, HER1 (EGRF) and HER4, visually in that order. HER-2 and HER-4 are absent in OVCAR-3 and SKOV-3, respectively. Qualitative phosphotyrosine levels were proportional to HER3 expression for each cell line, but HER3 activation was specifically detected primarily in OVCAR-8, PEO-4, and PEO-6 cells. HER3 gene silencing was efficiently performed in all lines studied, although only OVCAR-3 cells showed cell growth reduction by gene silencing.

Conclusões

Among the 7 tumor cell lines analyzed, only OVCAR-3 cells overexpressing HER3 were sensitive to the shRNA gene silencing technique, which indicates these receptors play a physiological role for cell proliferation and they can be targeted for the development of personalized drugs.

Financiador do resumo

CNPq

Palavras Chave

epithelial receptors; cell signalling; gynecologic tumors