CPLX2在30例肝癌组织的表达及其对体外细胞增殖与侵袭的影响

目的 检测人复合素2(CPLX2)在肝癌组织中的表达,探讨其对肝癌细胞增殖与侵袭的影响。 方法 采用RT-qPCR和Western blotting方法检测30例配对肝癌组织和癌旁组织中CPLX2 mRNA和蛋白水平的表达。利用Lipofectamine 2000转染两条CPLX2小分子干扰RNA(siRNA)至肝癌Huh7细胞,RT-qPCR和Western blotting方法验证转染后的干扰效率。细胞实验分4组:空白组、对照siRNA组,CPLX2 siRNA1组和CPLX2 siRNA2组。噻唑蓝(MTT)法和Transwell实验检测细胞增殖和侵袭能力。采用成组t检验分析siRNA干扰效果和细胞侵袭能力的差异,双因素方差分析法两两多重比较各组细胞增殖能力的差异。 结果 CPLX2在肝癌组织(22.69±14.78)中的表达高于癌旁组织(4.03±2.65),差异有统计学意义(t=5.941, P<0.001)。CPLX2 mRNA在两siRNA干扰组的表达量分别为0.34±0.02和0.48±0.01,均低于对照siRNA组(0.88±0.02)和空白组(1.00±0.05),差异有统计学意义(P<0.001),mRNA和蛋白水平均显示siRNA干扰效果较好。MTT实验证实CPLX2两siRNA干扰组的细胞增殖能力低于对照siRNA组(P<0.001)和空白组(P<0.001)。Transwell migration实验显示每个检测视野CPLX2 siRNA1组细胞的穿膜细胞数(37.0±2.0)和CPLX2 siRNA2组细胞的穿膜细胞数(46.3±2.5)低于空白组(88.0±2.0)和对照siRNA组(77.0±4.4)。Transwell invasion实验结果显示,每个检测视野CPLX2 siRNA1组细胞的穿膜细胞数(29.7±2.5)和CPLX2 siRNA2组细胞的穿膜细胞数(41.0±2.6)低于与空白组(74.7±3.1)和对照siRNA组(68.7±1.5),差异有统计学意义(P<0.001)。 结论 CPLX2在肝癌组织中表达升高,下调其表达可抑制肝癌细胞Huh7的增殖和侵袭,CPLX2可能在促进肝癌的发生发展过程发挥重要作用。

Expression of CPLX2 and its in vitro effects on the proliferation migration and invasion of hepatocellular carcinoma cells

Objective To investigate the expression of Complexin 2(CPLX2)in human hepatocellular carcinoma(HCC)and its effects on the proliferation and invasion of Huh7 cells. Methods The mRNA and protein expressions of CPLX2 in 30 HCC tissues and adjacent tissues were detected with RT-qPCR and Western blotting. Two CPLX2 small interfering RNA(siRNA)were transfected into Huh7 cells mediated by Lipofectamine 2000. The transfection efficiency of siRNA was confirmed by RT-qPCR and Western blotting. The cells were divided into 4 groups: blank group, siRNA control group, CPLX2 siRNA group 1 and CPLX2 siRNA group 2. The cell proliferation and migration were determined with methyl thiazol tetrazolium(MTT)and Transwell assay. The siRNA interference effects and cell invasion were analyzed with group t test. The cell proliferation in each group was compared with two-way ANOVA. Results The expression of CLPX2 was higher in HCC tissues than in adjacent tissues ［(22.69±14.78) vs(4.03±2.65), t=5.941, P<0.001］. The mRNA expression of CPLX2 was(0.34±0.02)in CPLX2 siRNA group 1 and(0.48±0.01)in CPLX2 siRNA group 2, which was lower than that in the siRNA control group(0.88±0.02)and blank group(1.00±0.05), and the difference was statistically significant(P<0.05), showing good siRNA interference effects. MTT assay confirmed the cell proliferation was lower in CPLX2 siRNA groups 1 and 2 than in the control group(P<0.001)and blank group(P<0.001). Transwell migration showed the number of penetrating cells in each detection field was(37.0±2.0)in CPLX2 siRNA group 1 and(46.3±2.5)in CPLX2 siRNA group 2, which was lower than that in the blank group(88.0±2.0)and siRNA control group(77.0±4.4). Transwell invasion showed the number of penetrating cells was(29.7±2.5)in CPLX2 siRNA group 1 and(41.0±2.6)in CPLX2 siRNA group 2, which was lower than that in the blank group(74.7±3.1)and siRNA control group(68.7±1.5), and the difference was statistically significant(P<0.001). Conclusion CPLX2 is highly expressed in HCC tissues and knockdown of CPLX2 can inhibit the proliferation and migration of Huh7 cells, indicating that CPLX2 may play an important role in HCC tumorigenesis and progression.