Cx43基因修饰对人肝癌SMMC-7721细胞增殖和迁移能力的影响

Effect of Cx43 gene on proliferation and migration of human hepatocellular carcinoma cell line SMMC-7721

目的探讨Cx43基因对人肝癌SMMC-7721细胞增殖、细胞周期及细胞迁移等方面的作用。方法采用LipofectamineTM2000转染的方法将重组表达质粒p Bud CE4.1-Cx43转染人肝癌SMMC-7721细胞(实验组),对照组细胞仅转染空载质粒p Bud CE4.1,空白组为未转染的SMMC-7721细胞。通过RT-PCR、Western blot检测转染后Cx43的mRNA和蛋白的表达情况,划痕荷载染料传输实验检测细胞间通讯功能,流式细胞仪检测细胞周期并通过CCK-8增殖实验、细胞划痕损伤实验、小室侵袭实验评价转染Cx43基因对人肝癌SMMC-7721细胞增殖、迁移以及侵袭能力的影响。结果实验组Cx43 mRNA和蛋白表达高于对照组和空白组,差异有统计学意义(均P<0.05)。实验组细胞传递的荧光强度高于对照组和空白组(P<0.05),实验组细胞周期被阻滞在G0/G1期,S期细胞数减少(P<0.05)。实验组与对照组和空白组比较,SMMC-7721细胞的增殖受到抑制(P<0.05),尤以72小时为著。实验组迁移能力下降,细胞侵袭能力下降,差异有统计学意义(P<0.05)。结论转染Cx43基因能够抑制人肝癌SMMC-7721细胞迁移的能力,并可以抑制肿瘤细胞增殖。

Objective To investigated the effect of Cx43 gene on the proliferation and migration of human hepatocellular carcinoma cell line SMMC-7721. Methods Recombination vector p Bud CE4. 1-Cx43 was transfected into human hepatocellular carcinoma SMMC-7721 cells with LipofectamineTM2000. The expression of Cx43 was detected by RTPCR and Western blot. Flow cytometry was used to examine cell cycle. Cell gap junction communication was measured by scrape-loading and dye transfer（ SLDT）. The effects of p Bud CE4. 1-Cx43 transfection on the proliferation,migration and invasion of SMMC-7721 cell line were evaluated by CCK-8,wound closure assay,chemotactic migration and cell invasive experiment,respectively. Results Compared with control group and blank group,mRNA and protein levels were significantly increased in experimental group transfected with p Bud CE4. 1-Cx43（ both P〈0. 05）. The cell cycle was arrested at G0/ G1 phase and cell number in S phase was decreased（ P〈0. 05）. Compared with control group and blank group,the cell proliferation in experimental group was inhibited significantly（ P〈0. 05）,especially at 72 h. The migration and invasion ability of cells in experimental group was decreased significantly（ P〈0. 05）. Conclusion p Bud CE4. 1-Cx43 transfection may decrease the migration,invasion and proliferation ability of human hepatocellular carcinoma SMMC-7721 cells.