Effect On Proliferation And Apoptosis Of The Colon Cancer Cells COLO205by Inhibition Of PIAS3Expression

Objective:Detected the expression level of PIAS3in human colon cancer cell line SW480(from colon cancer tissue, Dukes B), SW620(from lymphatic metastasis of colon cancer tissue, Dukes C) and COLO205(from colon cancer peritoneal metastatic tissue, Dukes D), which has different tissue source and different stages of Dukes. Sieved the cell line which has a higher expression of PIAS3used for further transfection experiments. Down regulated the expression of PIAS3in colon cancer cell line by transfection of PIAS3specific hairpin RNA (shRNA), observe the effection on proliferation, apoptosis and cell cycle of colon cancer cells. Methods:The expression of PIAS3was detected in SW480, SW620and COLO205cell lines of human colon cancer by Real-Time PCR and Western blot. Plasmids include PIAS3shRNA and scrambled control sequence were transfected into colon cancer cell lines mediated by cationic liposome. Cells differentiated into PIAS3shRNA transfection group (transfected with PIAS3shRNA plasmid), unrelated sequence transfection group (transfected with scrambled control sequence plasmid) and control group (without transfection). The expression of PIAS3mRNA and protein in different cancer cells were detected by Real-Time PCR and Western blot after transfected with different plasmid. Observe the proliferation of human colon cancer cells by MTT and the apoptosis and cycle of the cells were detected by FACS. Results:PIAS3was observed in SW480,SW620and COLO205cells by Real-Time PCR and Western blot, and the level of expression in COLO205cells was significantly high than the others. Therefore, COLO205was used for further shRNA intervention experiment. Compared with the Control group and unrelated sequence transfection group, the expression of PIAS3protein and mRNA had a significant decrease, the proliferation of COLO205cells was increased, and there was a sharply decrease of the COLO205cells in G1,G2phase, most of the cells arrested in S phase, the apoptosis rate significantly descreased. Conclusions:PIAS3has a positive expression in SW480, SW620and COLO205in three human colon cancer cell lines. Knockdown of PIAS3expression can promote the proliferation and inhibit apoptosis of colon cancer cells. The results that PIAS3can be used as a tumor suppressor has been replenished in this experiment。...