关键词: 结直肠癌, WIPI2, 慢病毒载体

Abstract:

WD repeat domain， phosphoinositide interacting 2 （WIPI2） is overexpressed in colorectal cancer （CRC）， suggesting that WIPI2 may be closely related to CRC progression， but the precise mechanism remain unclear. Therefore， the construction of a CRC cell model with stable overexpression of WIPI2 can provide a powerful experimental tool for in-depth study of the effect mechanism of WIPI2 in CRC and discovery of potential therapeutic targets for CRC. In this study， lentiviral vectors overexpressing WIPI2 were designed according to the human WIPI2 gene sequence and infected with CRC cells to evaluate the effect of the recombinant lentiviral vectors in enhancing the expression of WIPI2 protein. The sequence of the recombinant lentiviral vector was confirmed to be consistent with the target sequence of WIPI2 by gene sequencing， and the viral titer of the recombinant lentivirus was measured after viral packaging using 293T， and the titer of GL190-WIPI2 was 2.16×10⁸ TU·mL^-1； the expression level of WIPI2 protein in the overexpression of WIPI2 stably-transfected cell line was obviously up-regulated， and the lentiviral vector for overexpressing WIPI2 was successfully constructed and established. A lentiviral vector for WIPI2 overexpression was successfully constructed， and a stable transient CRC cell line overexpressing WIPI2 gene was established， which provides a cell model for studying the role of WIPI2 in CRC.

Key words: colorectal cancer, WIPI2, lentiviral vector