摘要
目的探讨虎杖苷对肝癌Huh-7细胞增殖、迁移及侵袭的影响及其可能作用机制。方法体外培养人肝癌细胞Huh-7,随机分组:对照组、低剂量虎杖苷组、中剂量虎杖苷组、高剂量虎杖苷组;应用克隆形成实验、CCK-8法、划痕实验与Transwell实验分别检测细胞增殖、迁移及侵袭;q RT-PCR法检测肝癌组织、癌旁组织与Huh-7细胞中miR-877-5p的表达量;E-cadherin、N-cadherin蛋白含量由Western blot法检测。结果相较于对照组,虎杖苷剂量依赖性的抑制细胞克隆形成数(分别为103.58±9.23、50.89±4.08,F=103.107,P=0.000),并通过促进E-cadherin蛋白表达(分别为0.16±0.02、0.55±0.05,F=187.778,P=0.000)抑制N-cadherin蛋白表达(分别为0.67±0.05、0.23±0.02,F=203.048,P=0.000)抑制细胞迁移(分别为71.96±5.94、28.12±2.54,F=164.192,P=0.000)和侵袭(分别为123.45±12.11、59.56±5.35,F=97.705,P=0.000);此外,虎杖苷刺激组细胞高表达miR-877-5p(分别为1.00±0.00、2.96±0.24,F=250.276,P=0.000),且成剂量依赖性。肝癌组织中miR-877-5p的表达量相较于癌旁组织显著降低(分别为1.00±0.08、0.32±0.03,t=55.712,P=0.000);miR-877-5p过表达相较于对照组被证实抑制肝癌细胞细胞克隆形成数(分别为108.41±11.37、59.33±5.12,t=11.808,P=0.000)、侵袭(分别为122.77±10.81、64.46±5.94,t=14.182,P=0.000)及迁移能力(分别为72.12±5.58、34.56±3.17,t=17.558,P=0.000);相较于虎杖苷+anti-miR-NC组,敲低虎杖苷刺激的肝癌细胞中miR-877-5p的表达可以逆转虎杖苷诱导的细胞克隆形成数(分别为48.52±4.50、89.23±7.88,t=13.459,P=0.000)、迁徙(分别为26.88±2.54、59.43±5.17,t=16.952,P=0.000)和侵袭抑制(分别为55.07±4.58、104.03±10.15,t=13.190,P=0.000)。结论虎杖苷可通过上调miR-877-5p表达而破坏肝癌细胞增殖、迁移及侵袭能力。
Objective To explore the effect of polydatin on the proliferation,migration and invasion of liver cancer Huh-7 cells and its possible mechanism.Methods Human hepatocellular carcinoma cells Huh-7 were cultured in vitro and randomly divided into groups:control group,polydatin low-dose group,polydatin medium-dose group,and polydatin high-dose group.CCK-8 method,plate clone formation test,scratch test and Transwell test were used to detect cell proliferation,clone formation,migration and invasion.qRT-PCR was used to detect the expression of miR-877-5p in liver cancer tissues,adjacent tissues and Huh-7 cells.Subsequent experimental groups:miR-NC group,miR-877-5p group,polydatin+anti-miR-NC group,polydatin+anti-miR-877-5p group.The above methods were used to detect cell proliferation,clone formation,migration and invasion.Western blot was used to detect the expression of E-cadherin and N-cadherin protein.Results Compared with the control group,polydatin dosedependently inhibited the number of cell clone formation(103.58±9.23 vs 50.89±4.08,respectively,F=103.107,P<0.05),suppressed E-cadherin protein expression(0.16±0.02 vs 0.55±0.05,respectively,F=187.778,P=0.000),promoted N-cadherin protein expression(0.67±0.05 vs 0.23±0.02,respectively,F=203.048,P=0.000),inhibited cell migration(71.96±5.94 vs 28.12±2.54,respectively,F=164.192,P=0.000),and invasion(123.45±12.11 vs59.56±5.35,respectively,F=97.705,P=0.000).In addition,cells in the polydatin-stimulated group highly expressed miR-877-5p(1.00±0.00 vs 2.96±0.24,respectively,F=250.276,P=0.000)in a dose-dependent manner.Compared with adjacent tissues,the expression of miR-877-5p was significantly lower in hepatocellular carcinoma tissues(1.00±0.08 vs 0.32±0.03,respectively,t=55.712,P=0.000).Compared with the miR-NC group,miR-877-5p overexpression was shown to inhibit the number of clone formation in Huh-7 cells(108.41±11.37 vs 59.33±5.12,respectively,t=11.808,P=0.000),invasion(122.77±10.81 vs 64.46±5.94,respectively,t=14.182,P=0.000)and migration ability(72.1
作者
戴鹏
戴丽
李德龙
张伟
DAI Peng;DAI Li;LI De-long;ZHANG Wei(Department of Comprehensive Internal Medicine,Shanxi Cancer Hospital,Taiyuan,Shanxi Province 030013,China;Department of Dermatology,Shanxi Staff Medical College,Taiyuan,Shanxi Province 030012,China;Department of Emergency,Shanxi Cancer Hospital,Taiyuan,Shanxi Province 030013,China)
出处
《解剖学研究》
CAS
2023年第5期441-446,共6页
Anatomy Research
