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目的检测TIMP-3在人肝癌组织中的表达,研究其对肝癌细胞增殖、凋亡和周期等细胞功能的影响。方法免疫组织化学检测80例肝癌及60例癌旁组织中TIMP-3表达。建立以真核表达载体介导的过表达TIMP-3的肝癌细胞株HepG2,用Western Blot检测细胞内TIMP-3蛋白的表达,通过MTS实验和流式细胞仪研究TIMP-3对HepG2肝癌细胞各生物学行为的影响。结果与癌旁组织相比,肝癌组织TIMP-3的阳性表达率则显著下降(40.00%vs.61.67%)。体外试验显示转染了pCMV6-AC-GFP/TIMP-3的HepG2肝癌细胞TIMP-3蛋白高表达,而对照组的肝癌细胞则TIMP-3低表达。相对对照组,试验结果显示TIMP-3不仅使肝癌细胞HepG2的增殖力减弱,且诱导其发生凋亡,并使细胞周期停滞于G2/M期。结论 TIMP-3在肝癌组织中低表达,参与肝癌的生成,并影响了肝癌细胞的增殖、凋亡和周期。
Objective To detect the expression of TIMP-3 in human hepatocellular carcinoma (HCC) and to study its effects on the proliferation, apoptosis and cell cycle of hepatocellular carcinoma cells. Methods Immunohistochemistry was used to detect the expression of TIMP-3 in 80 cases of HCC and 60 cases of paracancerous tissues. To establish a hepatoma cell line HepG2 overexpressing TIMP-3 mediated by eukaryotic expression vector and detect the expression of TIMP-3 protein by Western Blot. To study the effect of TIMP-3 on the expression of TIMP-3 in HepG2 hepatocellular carcinoma by using MTS assay and flow cytometry The impact of biological behavior. Results The positive rate of TIMP-3 in HCC tissues was significantly lower than that in paracancerous tissues (40.00% vs.61.67%). In vitro experiments showed that the expression of TIMP-3 protein in HepG2 hepatoma cells transfected with pCMV6-AC-GFP / TIMP-3 was high, while that in control cells was low. Compared with the control group, TIMP-3 not only reduced the proliferation of HepG2 hepatocarcinoma cells, but also induced the apoptosis of cells and arrested the cell cycle in G2 / M phase. Conclusions TIMP-3 is low expressed in HCC tissues and is involved in the generation of HCC and affects the proliferation, apoptosis and cycle of HCC cells.