Network analysis on Rab7a gene in breast cancer MDA-MB-231 cells / 吉林大学学报(医学版)

ABSTRACT

Objective: To investigate the changes of expressions of the related genes in the triple-negative breast cancer (TNBC) MDA-MB-231 cells after knockout of Rab7a gene, and to elucidate the roles of Rab7a-related genes in TNBC. Methods: The breast cancer ZR-75-30, MCF-7, T-47D, MDA-MB-231 and HCC-1937 cells in the logarithmic phase were selected. The expression levels of Rab7a protein in the breast cancer ZR-75-30, MCF-7, T-47D, MDA-MB-231, and HCC-1937 cells were detected by Western blotting method. The Rab7a gene in the MDA-MB-231 cells was knockout with lentivirus and the cells were divided into negative control group and Rab7a knockout group. According to the four different knockout Rab7a sequences, the Rab7a knockout group was divided into KD1 group, KD2 group, KD3 group and KD4 group. The knockout efficiencies of Rab7a gene in the MDA-MB-231 cells in various groups were detected by qPCR method, the differential genes in the highest knockout efficiency group (KD2 group) and negative control group were detected by full gene expression microarray, and the interaction between Rab7a gene and other genes was analyzed by integrated pathway analysis (IPA) software. Results: The Rab7a protein was expressed in the TNBC MDA-MB-231 cells. Compared with negative control group, the knockout efficiency of Rab7a gene in the MDA-MB-231 cells in KD2 group was the highest (Pcontrol group, there were 634 differential genes in the MDA-MB-231 cells in KD2 group, the number of up-regulated genes and down-regulated genes were increased (P < 0. 01); the differential gene analysis results showed that Rab7a knockout had the relationships with cancer, cell survival, cell cycle, cell growth, and migration. The expressions of eIF4F, IRS1 and RPS6KB1 in the Rab7a IPA network diagram were down-regulated, and the expressions of PRKAA1, IKBKE, VEGFA, and transcription factor ATF2 were up-regulated. Concuson Rab7a gene plays a role as a gene network in the TNBC MDA-MB-231 cells. The Rab7a gene interacts with the multiple genes and participates in the pathological processes of cancer cells.