关键词: 甲磺酸盐及紫外线敏感性81号基因, 肝细胞癌, 增殖, 凋亡, 小干扰RNA

Abstract:

Objective To study the effects of methyl methanesulfonate and UV sensitive gene clone 81（Mus81）silencing on proliferation and apoptosis of human hepatocellular carcinoma Hep3B cells. Methods Lenti⁃virus⁃mediated small interfering RNA（siRNA）was employed to inhibit Mus81 expression and real⁃time quantitativepolymerase chain reaction（Real⁃time PCR）was used to determine the inhibition efficiency of Mus81 expression.Growth curve was established by the methylthiazol tetrazolium（MTT）assay. The plate cloning experiment，propidiumiodide（PI）and Annexin V⁃APC staining were used to detect the proliferation，cell cycle distribution and apoptosisof Hep3B cell line. Results The efficiency of lentivirus infection was higher than 80%，and the efficiency of Mus81gene interference was 76.42%（P＜0.001）. The results of MTT growth curve and plate cloning experiment showedthat the growth rate of the Hep3B cell line silenced by Mus81 was decreased significantly（P < 0.001），the proliferationability was also significantly weakened（number of cell clones：（2 ± 1）vs.（54 ± 3），P < 0.001）. Annexin V⁃APCand PI staining showed that the apoptosis rate of Mus81 silenced Hep3B cell line was significantly increased［（20.87 ± 0.82）% to（0.69 ± 0.09）%，P < 0.001］and the proportion of cells in G1 phase was slightly decreased［（33.77 ± 1.61）% to（42.06 ± 0.40）%，P < 0.05］. Conclusion Mus81 gene silencing can significantly inhibit thegrowth and proliferation of human liver cancer Hep3B cells and promote its apoptosis.

Key words: methyl methanesulfonate and UV sensitive gene clone 81, hepatocellular carcinoma, pro?liferation, apoptosis, small interfering RNA