Hsa-miR-4282对肝癌细胞系SMMC-7721生长的影响

目的探讨Hsa-miR-4282在肝癌细胞系SMMC-7721中的表达及其对细胞生长的影响。方法采用实时荧光定量PCR法检测Hsa-miR-4282在人正常肝上皮细胞系HL-7702和人肝癌细胞系MHCC97-H、SMMC-7721,以及 20例肝癌组织及其相应癌旁组织中的表达。采用瞬时转染法将Hsa-miR-4282 mimics(上调组)和Hsa-miR-4282 inhibitor(下调组)分别转染肝癌SMMC-7721细胞,上调组和下调组分别设置相应阴性对照组。转染后采用MTT法检测细胞增殖能力,平板克隆形成实验检测细胞克隆形成能力,流式细胞仪检测细胞凋亡能力。结果 Hsa-miR-4282在肝癌组织、肝癌细胞MHCC97-H及肝癌细胞SMMC-7721中的表达均低于癌旁组织及正常肝细胞HL-7702(P<0.05)。MTT实验结果显示,Hsa-miR-4282上调后肝癌SMMC-7721细胞的OD值低于其阴性对照组,而下调后OD值高于其阴性对照组(P<0.05)。平板克隆形成实验显示,下调组的细胞克隆数高于其阴性对照组(240±7)个 vs (191±10)个,P=0.005)],而上调组细胞克隆数低于基阴性对照组(146±10)个 vs (193±12)个,P=0.013)]。流式细胞仪检测结果显示,Hsa-miR-4282上调组细胞凋亡率较其阴性对照组升高(23.89±1.89)% vs(16.6±1.14)%,P=0.009)],下调组细胞凋亡率较期阴性对照组降低(14.98±0.46)% vs (17.79±0.73)%,P=0.010]。结论 Hsa-miR-4282上调可抑制肝癌SMMC-7721细胞增殖,促进细胞凋亡,可能与肝癌的发病机制有关。

Effect of Hsa-miR-4282 on the growth of hepatoma cell line SMMC-7721

Objective To investigate the expression of Hsa-miR-4282 in hepatocellular carcinoma cell line SMMC-7721 and its effect on cell growth. Methods The expression of Hsa-miR-4282 in human normal liver epithelial cell line HL-7702，human hepatoma cell lines MHCC97-H and SMMC-7721，20 cases of primary liver cancer tissues and their corresponding adjacent tissues were detected by quantitative real-time PCR. The SMMC-7721 cells were transfected with Hsa-miR-4282 mimics（up-regulation group） or Hsa-miR-4282 inhibitor（down-regulation group） by transient transfection method，and negative control group were set in the up-regulated group and the down-regulated group，respectively.After transfection，the cell proliferation ability was detected by MTT assay，the cell clone formation ability was detected by plate cloning assay，and the apoptosis ability was detected by flow cytometry. Results The expres-sion of Hsa-miR-4282 in liver cancer tissues，liver cancer cells MHCC97-H and liver cancer cells SMMC-7721 were lower than that in adjacent tissues and normal liver cells HL-7702 （P<0.05）.The MTT assay showed that the OD value of liver cancer SMMC-7721 cells up-regulated by Hsa-miR-4282 was lower than that of the negative control group，but the OD value of liver cancer SMMC-7721 cells down-regulated by Hsa-miR-4282 was higher than that of the control group.Plate cloning experiments showed that the number of cell clones in the down-regulated group was higher than that in the negative control group ［（240±7） vs （191±10），P=0.005）］，while the number of cell clones in the up-regulated group was lower than that in the control group ［（146±10） vs （193±12），P=0.013）］.The results of flow cytometry showed that the apoptosis rate of Hsa-miR-4282 up-regulated group was higher than that of the control group ［（23.89±1.89）% vs （16.6±1.14）%，P=0.009）］，while the rate in Hsa-miR-4282 down-regulated group was lower than that of the control group ［（14.98±0.46）% vs （17.79±0.73）%，P=0.010］. Conclusions  Up-regulation of Hsa-miR-4282 can inhibit the proliferation of liver cancer SMMC-7721 cells and promote cell apoptosis，which may be related to the pathogenesis of liver cancer.