Overexpression of TIMP3 Affects Biological Functions of Liver Cancer Cells
CAO Li1, WU Fei2, WANG Xiaofei3*, QIN Yannan1*
The role of TIMP3 (tissue inhibitor of metalloproteinase 3) in hepatocellular carcinoma is still unclear. This study aims to explore the expression of TIMP3 in HCC (hepatocellular carcinoma) cell lines, and its effect on cell proliferation, apoptosis and cell cycle in 97H and HUH7. The expression of TIMP3 in HCC cell lines (97H, 97L and HUH7) and normal human hepatocytes (LO2) was detected by qRT-PCR and Western blot. The overexpressing plasmids were transfected into 97H and HUH7 cells, and the expression of TIMP3 in the transfected cells was examined by qRT-PCR and Western blot. The effect of TIMP3 overexpression on the proliferation of 97H and HUH7 cell was detected by MTT assay and further monitored by cell imaging multi-mode reader. In addition, the effect of TIMP3 overexpression on apoptosis, cell cycle, and cell migration was detected by flow cytometry and wound-healing assay. Finally, GSEA (gene enrichment analysis) and IntAct database were used to explore the biological functions of TIMP3 in HCC. The results showed that expression of TIMP3 in 97H, 97L and HUH7 cells was lower than that in LO2 cells. The levels of RNA and protein in 97H and HUH7 cells transfected with TIMP3 plasmid were significantly higher, and overexpression of TIMP3 could promote cell proliferation, promoting the transition from G1 phase to S phase, and inhibit cell apoptosis (downregulation of Bax expression, upregulation of Bcl-2 and Bcl-xl expression). All in all, TIMP3 overexpression significantly promotes the proliferation and inhibits cell apoptosis in HCC, and TIMP3-related signaling pathways and interacting proteins are closely involved in tumor progression. TIMP3 may be a potential diagnostic marker and therapeutic target for hepatocellular carcinoma.
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