GINS复合物4在肝细胞癌组织中的表达及其对肝癌细胞侵袭、增殖能力的影响

The expression of GINS complex 4 in hepatocellular carcinoma tissues and its effect on the invasion and proliferation of hepatocellular carcinoma cell

目的检测肝细胞癌组织中GINS复合物4(GINS4)蛋白的表达,观察其对肝癌细胞侵袭、增殖能力的影响。方法选取河南大学人民医院2019年9月至2020年12月60例肝细胞癌患者术中收集肝细胞癌组织及对应癌旁组织标本分别作为观察组和对照组,采用实时荧光定量聚合酶链反应(RT-qPCR)和蛋白质印迹法(Western blot)检测GINS4在肝细胞癌组织中的表达,使用特异性短发卡RNA(shRNA)构建低表达的肝癌细胞稳定株,将其分为实验组(shRNA组)和对照组(NC组);Transwell检测其对HepG2细胞的侵袭能力;集落形成实验和细胞计数试剂盒(CCK-8)检测其对HepG2细胞的增殖能力。组间比较采用配对样本t检验。结果RT-qPCR表明GINS4在肝细胞癌组织中表达量(2.89±0.11)高于癌旁组织表达量(1.34±0.15),差异有统计学意义(t=8.56,P<0.01)。Western blot显示在肝细胞癌组织中GINS4表达量高于癌旁组织的(1.54±0.76)倍,差异有统计学意义(t=2.67,P<0.05),shRNA转染HepG2细胞后,实验组GINS4表达量较对照组的表达量低(1.88±0.34)倍,差异有统计学意义(t=7.43,P<0.05);Transwell实验显示实验组穿孔细胞数目[(56.33±3.51)个/视野]低于对照组[(90.42±1.53)个/视野],差异有统计学意义(t=3.34,P<0.01);集落形成实验显示实验组细胞集落数量[(83.33±7.37)个]低于对照组[(345.00±16.64)个],差异有统计学意义(t=15.28,P<0.01);CCK-8实验显示在3 d的吸光度值实验组(1.43±0.11)低于对照组(2.15±0.15),差异有统计学意义(t=6.52,P<0.01)。结论在肝细胞癌组织中GINS4表达量高于癌旁组织,降低GINS4的表达能够显著抑制肝癌细胞侵袭及增殖能力。

Objective To detect the expression of GINS complex subunit 4(GINS4)protein in hepatocellular carcinoma tissues and observe its effect on the invasion and proliferation of hepatocellular carcinoma cells.Methods From September 2019 to December 2020 in our hospital,60 patients with hepatocellular carcinoma were collected during surgery and hepatocellular carcinoma tissues and corresponding para-cancerous tissue specimens were selected as the observation group and the control group.Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)and Western blotting was used to detect the expression of GINS4 in hepatocellular carcinoma tissues,use specific short hairpin RNA(shRNA)to construct low-expressing stable hepatocellular carcinoma cell lines,and divide them into experimental groups(shRNA group)and the control group(NC group);Transwell tested its ability to invade HepG2 cells;colony formation test and cell counting kit(CCK-8)test tested its proliferation ability on HepG2 cells.Comparison between groups is by paired sample t test.Results RT-qPCR showed that the expression of GINS4 in hepatocellular carcinoma tissues(2.89±0.11)was higher than that in adjacent tissues(1.34±0.15),and the difference was statistically significant(t=8.56,P<0.01).Western blot showed that the expression of GINS4 in hepatocellular carcinoma tissue was(1.54±0.76)times higher than that in adjacent tissues,and the difference was statistically significant(t=2.67,P<0.05).After shRNA transfection of HepG2 cells,GINS4 in the experimental group The expression level was(1.88±0.34)times lower than that of the control group,and the difference was statistically significant(t=7.43,P<0.05);the Transwell experiment showed that the number of perforated cells in the experimental group was(56.33±3.51)compared with the control group(90.42±1.53)cells were significantly reduced,and the difference was statistically significant(t=3.34,P<0.01);the colony formation experiment showed that the number of cell clusters in the experimental group(83.33±7.37)was significantly less than that in the control group(345.00±16.64).The difference was statistically significant(t=15.28,P<0.01);CCK-8 experiment showed that the 3 d absorbance of the experimental group was(1.43±0.11)lower than that of the control group(2.15±0.15),and the difference was statistically significant(t=6.52,P<0.001).Conclusion The expression of GINS4 in hepatocellular carcinoma tissues is higher than that in adjacent tissues.Reducing the expression of GINS4 can significantly inhibit the invasion and proliferation of hepatocellular carcinoma cells.