干扰miR-21靶向调控PDCD4、PTEN和TPM1抑制肝癌细胞增殖和侵袭能力

Inhibition of miR-21 inhibits proliferation and invasion of liver cancer cells by targeting PDCD4, PTEN and TPM1

目的运用构建好的重组慢病毒载体系统PLVX-MIR21-RNAi分别感染高、低侵袭性肝癌细胞HCCLM3和HepG2,并观察miR-21下调对其生长增殖和侵袭转移特性的影响及其分子机制研究。方法采用qPCR、CCK-8、Transwel肿瘤侵袭和Westermn blot 实验分别检测干扰miR-21表达对HCCLM3和HepG2细胞miR-21表达量、增殖抑制率、侵袭转移特性和PTEN、PDCD4和TPM1蛋白的表达量影响。结果HCCLM3转染组miR-21的表达量低于HepG2转染组(P<0.05);24 h 36 h、48 h时,HCCLM3未处理组增殖能力均明显高于HepG2未处理组(P<0.01),而HCCLM3转染组增殖抑制率均高于HepG2转染组(P<0.05);HCCLM3转染组细胞侵袭抑制率高于HepG2转染组(P<0.05);HCCLM3未处理组PTEN、PDCD4和TPM1蛋白表达低于HepG2未处理组(P<0.01)。此外,下调miR-21后,HCCLM3转染组PTEN PDCD4和TPM1蛋白表达水平均高于HepG2转染组(P <0.01)。结论miR-21 下调能有效抑制HCCLM3和HepG2细胞生长增殖和侵袭转移能力,且与其靶向调控PDCD4、PTEN、TPM1基因有关。

Objective PLVX-shRNA2 lentiviral vector system was established and transfected into high/ low-invasion liver cancer cells HCCLM3 and HepG2;to investigate the proliferation, invasive ability effects of Inhibition of miR-21 on these cells, and to clarify the underlying mechanisms. Methods Quantitative PCR ( qPCR) assay was used to examine the miR-21 levels. Cell counting kit-8 (CCK-8) assay was performed to observe cell viabilities. Transwell assay was conducted to evalu-ate the invasion potential. Western blot assay was used to determine the protein expression of PTEN, PDCD4 and TPM1. Results MiR-21 level was significantly lower in HCCLM3-PLVX-anti-miR-21 group compared to HepG2-PLVX-anti-miR-21 (P<0. 05). The proliferation of HCCLM3-untreated cells was significantly higher compared to HepG2-untreated cells at 24 h, 36 h and 48 h, respectively (P <0.01). The proliferation inhibition rates of HCCLM3-PLVX-anti-miR-21 cells were higher compared to HepG2-PLVX-anti-miR-21 cells at 24 h, 36 h and48 h (P <0.05). The inva-sive inhibition rate in HCCLM3-PLVX-anti-miR-21 cells was significantly higher compared with HepG2-PLVX-anti-miR-21 cells (P <0.05). The protein expression of PTEN, PDCD4 and TPM1 in HCCLM3-untreated cells were lower than HepG2-untreated cells (P <0.01). The protein expression of PTEN, PDCD4 and TPM1 in HCCLM3一PLVX-anti-miR-21 ellls were higher than HepG2-PLVX-anti-miR-21 cells by Western blot (P <0.01 ). Conclusion Down-regulation of miR-21 can effectively inhibit the growth, invasion and metastasis of HCCLM3 and HepG2 cells, and it is related to the targeted regulation of PDCD4, PTEN and TPM1 genes.