微小RNA-127-3p在乙型肝炎病毒相关性肝癌中的表达意义及对HBV阳性肝癌细胞增殖迁移侵袭的影响

Expression of miR-127-3p in hepatitis B virus associated hepatocellular carcinoma and its effect on proliferation,migration and invasion of HBV positive hepatocellular carcinoma cells

目的:探讨微小RNA-127-3p(miR-127-3p)在乙型肝炎病毒(HBV)相关性肝癌中的表达意义及其对HBV阳性肝癌细胞增殖、迁移和侵袭的影响。方法:采用实时荧光定量聚合酶链反应(RTFQ-PCR)检测2018年12月至2020年12月在医院手术切除的64例HBV相关性肝癌患者肝癌组织和癌旁组织中miR-127-3p表达量。将对数生长期肝癌HepG2细胞分别以10、20、50 pmol的miR-127-3p mimic进行转染,同时设置对照组(以0 pmol mimic进行转染),培养48 h后采用RTFQ-PCR检测各转染组细胞中miR-127-3p表达量,MTT法检测各转染组细胞增殖情况,细胞划痕实验检测细胞迁移能力,Transwell实验检测细胞侵袭能力。结果:肝癌组织miR-127-3p表达量(0.26±0.03)低于癌旁组织(0.53±0.06)(P<0.05)。miR-127-3p mimic转染组(10、20、50 pmol)与对照组肝癌HepG2细胞中miR-127-3p表达量分别为(0.87±0.09)、(1.23±0.13)、(1.51±0.16)、(0.28±0.04),总体比较差异有统计学意义(P<0.05);与对照组比较,miR-127-3p mimic(10、20、50 pmol)转染组miR-127-3p表达量更高(P<0.05);miR-127-3p mimic转染组中miR-127-3p表达量随转染浓度的升高而升高(P<0.05)。miR-127-3p mimic转染组(10、20、50 pmol)与对照组肝癌HepG2细胞增殖率分别为[(51.58±5.24)%、(25.42±2.55)%、(16.23±1.65)%、(78.67±7.88)%],细胞迁移距离分别为[(28.26±2.84)mm、(23.17±2.25)mm、(18.32±1.83)mm、(33.87±3.41)mm],穿膜细胞数分别为[(122.54±12.26)个、(101.26±10.45)个、(46.15±4.78)个、(131.23±13.24)个],总体比较差异均有统计学意义(P<0.05);与对照组比较,miR-127-3p mimic(10、20、50 pmol)转染组细胞增殖率更低,细胞迁移距离更短,穿膜细胞数更少(P<0.05);miR-127-3p mimic转染组细胞增殖率随转染浓度的升高而降低,细胞迁移距离随转染浓度的升高而变短,穿膜细胞数随转染浓度的升高而减少(P<0.05)。结论:miR-127-3p在HBV相关性肝癌患者中呈低表达,其可抑制HBV阳性肝癌HepG2细胞增殖、迁移和侵袭。

Objective:To investigate the significance of microRNA-127-3p(miR-127-3p)expression in hepatitis B virus(HBV)-associated hepatocellular carcinoma and its effects on the proliferation,migration and invasion of HBV-positive hepatocellular carcinoma cells.Methods:Real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR)was used to detect miR-127-3p expression in liver cancer tissues and paracancerous tissues of 64 patients with HBV-associated hepatocellular carcinoma surgically resected in hospitals from December 2018 to December 2020.Logarithmic growth phase hepatocellular carcinoma HepG2 cells were transfected with 10,20,and 50 pmol of miR-127-3p mimic,while a control group(transfected with 0 pmol mimic)was set up.After 48 h of culture,RTFQ-PCR was used to detect the expression of miR-127-3p in each transfected group,and MTT assay was used to detect the proliferation of each transfected group.The cell proliferation was detected by RTFQ-PCR,the cell migration ability was detected by MTT,and the cell invasion ability was detected by Transwell assay.Results:The miR-127-3p expression in hepatocellular carcinoma tissue(0.26±0.03)was lower than that in paraneoplastic tissue(0.53±0.06)(P<0.05).The miR-127-3p expression in miR-127-3p mimic transfected groups(10,20,and 50 pmol)and control hepatoma HepG2 cells were(0.87±0.09),(1.23±0.13),(1.51±0.16),(0.28±0.04),respectively,and the overall comparison was statistically significant(P<0.05).Compared with the control group,miR-127-3p mimic(10,20,and 50 pmol)transfected group had higher miR-127-3p expression(P<0.05).The miR-127-3p expression in miR-127-3p mimic transfected group increased with the increase of transfection concentration(P<0.05).The proliferation rates of hepatocellular carcinoma HepG2 cells in miR-127-3p mimic transfected group(10,20,and 50 pmol)and control group were[(51.58±5.24)%,(25.4±2.55)%,(16.23±1.65)%,(78.67±7.88)%],respectively.The cell migration distances were[(28.26±2.84)mm,(23.17±2.25)mm,(18.32±1.83)mm,(33.87±3.41)mm],and the numbers of membrane penetrating cells were[(122.54±12.26),(101.26±10.45),(46.15±4.78),(131.23±13.24)],all with statistically significant differences in the overall comparison(P<0.05).Compared with the control group,miR-127-3p mimic(10,20,and 50 pmol)transfected group had a lower cell proliferation rate,shorter cell migration distance,and less number of membrane penetrating cells(P<0.05).The miR-127-3p mimic transfected group had lower cell proliferation rate with increasing transfection concentration,shorter cell migration distance with increasing transfection concentration,and less number of membrane penetrating.The cell proliferation rate of miR-127-3p mimic transfected group decreased with the increase of transfection concentration,cell migration distance became shorter with the increase of transfection concentration,and the number of membrane penetration cells decreased with the increase of transfection concentration(P<0.05).Conclusion:The miR-127-3p was lowly expressed in HBV-associated hepatocellular carcinoma patients,and it inhibited the proliferation,migration,and invasion of HBV-positive hepatocellular carcinoma HepG2 cells.