摘要
[目的]探讨化疗药物诱导肝癌细胞凋亡的潜在机制。[方法]高通量测序检测顺铂处理后肝癌细胞HepG2的失调miRNA。过表达失调miRNA后,检测HepG2的凋亡水平。miRDB在线分析和RNAi筛选鉴定miRNA的潜在底物。[结果]顺铂处理后,HepG2的凋亡水平上升(18.23±3.01 vs 74.97±11.25,t=8.564,P<0.05)。过表达miR-301a-3p时,HepG2的凋亡水平上升(16.40±7.97 vs 56.12±9.37,t=5.590,P<0.05)。敲低miR-301a-3p时,HepG2的凋亡水平下降(21.34±3.34 vs 6.78±1.11,t=7.165,P<0.05)。敲低MDM4时,HepG2的凋亡水平上升(16.80±7.88 vs 72.85±10.70,t=7.302,P<0.05)。过表达MDM4时,HepG2的凋亡水平下降(19.11±5.02 vs 6.04±1.02,t=4.432,P<0.05)。过表达miR-301a-3p时,MDM4的mRNA水平和蛋白水平均下降(0.58±0.04 vs 0.11±0.01,t=19.74,P<0.05);而敲低miR-301a-3p时,MDM4的mRNA水平和蛋白水平均上升(0.58±0.04 vs 1.11±0.23,t=3.932,P<0.05)。肝癌组织中miR-301a-3p的表达水平显著低于癌旁组织(0.65±0.25 vs 0.40±0.04,t=5.949,P<0.05)。肝癌组织中MDM4的表达水平显著高于癌旁组织(0.036±0.025 vs 0.062±0.003,t=6.550,P<0.05)。同时,肝癌组织中miR-301a-3p和MDM4的表达水平呈现负相关(ρ=-0.385,P<0.05)。[结论]顺铂诱导肝癌细胞miR-301a-3p表达上调,miR-301a-3p通过靶向MDM4 mRNA的3’端非编码区,降解了MDM4 mRNA,减少了MDM4的蛋白表达,最终促进了肝癌细胞的凋亡。
[Objective]To investigate the potential mechanism of apoptosis induced by chemotherapeutic drugs in hepatocellular carcinoma cells.[Method]High-throughput sequencing was performed to detect the dysregulated miRNA of HepG2 in hepatocellular carcinoma cells after cisplatin treatment.apoptosis level of HepG2 was detected after overexpression of dysregulated miRNA.miRDB online analysis and RNAi screening were performed to identify potential substrates of miRNA.[Result]Apoptosis level of HepG2 increased after cisplatin treatment(18.23±3.01 vs 74.97±11.25,t=8.564,P<0.05).The apoptotic level of HepG2 increased upon overexpression of miR-301a-3p(16.40±7.97 vs 56.12±9.37,t=5.590,P<0.05).The apoptotic level of HepG2 decreased when miR-301a-3p was knocked down(21.34±3.34 vs 6.78±1.11,t=7.165,P<0.05).The apoptotic level of HepG2 increased when MDM4 was knocked down(16.80±7.88 vs 72.85±10.70,t=7.302,P<0.05).The apoptosis level of HepG2 decreased when overexpressing MDM4(19.11±5.02 vs 6.04±1.02,t=4.432,P<0.05).Both mRNA levels and protein levels of MDM4 decreased when miR-301a-3p was overexpressed(0.58±0.04 vs 0.11±0.01,t=19.74,P<0.05),while both mRNA levels and protein levels of MDM4 increased when miR-301a-3p was knocked down(0.58±0.04 vs 1.11±0.23,t=3.932,P<0.05).The expression level of miR-301a-3p was significantly lower in hepatocellular carcinoma tissues than in paraneoplastic tissues(0.65±0.25 vs 0.40±0.04,t=5.949,P<0.05).The expression level of MDM4 was significantly higher in hepatocellular carcinoma tissues than in paraneoplastic tissues(0.036±0.025 vs 0.062±0.003,t=6.550,P<0.05).Meanwhile,the expression levels of miR-301a-3p and MDM4 in hepatocellular carcinoma tissues showed a negative correlation(ρ=-0.385,P<0.05).[Conclusion]Cisplatin induced upregulation of miR-301a-3p expression in hepatocellular carcinoma cells.miR-301a-3p degraded MDM4 mRNA by targeting the non-coding region at the 3'end of MDM4 mRNA,reduced the protein expression of MDM4,and finally promoted apoptosis of hepatocellular carc
作者
倪淑斌
张建勋
徐星
金翌
杨静
NI Shu-bin;ZHANG Jian-xun;XU Xing;JIN Yi;YANG Jing(Hepatobiliary Surgery,Seventh People's Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200137,China)
出处
《生物技术》
CAS
2022年第3期332-337,共6页
Biotechnology
基金
上海中医药大学附属第七医院人才培养计划(BDX2022)
上海中医药大学“研究生创新培养专项”(Y2021045)。
