类胰岛素生长因子-1调控微小RNA-145对肝癌术后复发转移的影响

Effects of insulin like growth factor-1 on recurrence and metastasis of hepatocellular carcinoma after hepatectomy regulated through microRNA-145

目的:探讨肝切除术后类胰岛素生长因子-1(IGF-1)的表达变化,及其对微小RNA(miRNA,miR)-145的调控在肿瘤复发转移中的作用。方法:收集吉林大学中日联谊医院2016年至2017肝癌手术患者40例,检测术后血清IGF-1浓度变化;建立小鼠肝切除模型,检测术后血清IGF-1表达变化;Transwell检测IGF-1作用后HepG2细胞侵袭能力的变化以及miR-145、E盒结合锌指蛋白2(Zeb2)的表达变化;脂质体转染mimics和miR-145 inhibitor,检测HepG2细胞侵袭能力的变化。组间比较采用t检验分析两组。结果:患者术后30 d血清IGF-1浓度[(53.90±6.53)μg/L]高于术后1 d[(43.00±6.37)μg/L,t=-3.804,P<0.01],小鼠术后21 d血清IGF-1浓度[(74.00±4.73)μg/L]高于术后3 d[(57.00±6.87)μg/L,t=-8.046,P<0.01];Transwell显示处理组细胞侵袭能力[(165.67±10.01)个]明显高于对照组[(50.66±4.04)个,t=-20.880,P<0.01],miR-145表达(0.224±0.019)低于对照组(1.000±0.042,t=20.074,P<0.01);Zeb2表达(0.91±0.25)高于对照组(0.09±0.72,t=-53.959,P<0.01);转染抑制和增强miR-145表达后,miR-145 inhibitor组Zeb2蛋白表达和细胞侵袭数目[(1.33±0.38)、(259.00±14.10)个]较miR-145 mimics组[(0.26±0.19)、(68.00±4.58)个]明显增加(P<0.01)。结论:肝切除术后IGF-1表达增加,可能通过调控miR-145影响肿瘤的复发转移。

Objective To investigate the expression of insulin-like growth factor 1(IGF-1)after hepatectomy and its mechanism for the regulation of microRNA(miRNA,miR)-145 in tumor recurrence and metastasis.Methods The enzyme linked immunosorbent assay(ELISA)kit was used to detect the changes of serum IGF-1 in patients with hepatectomy after 1,3 and 30 days.The hepatectomy model of mice was established to detect the changes of serum IGF-1 after surgery of 3,7 and 21 days.Transwell invasion assay was used to detect the effect of IGF-1 on the invasion of HepG2 cells.The expression of miR-145 and Zeb2 was dectected by qPCR and Western blotting in mice treated with IGF-1.The effect of miR-145 on the invasion of HepG2 cells was examined by Transwell invasion assay.Results The serum of IGF-1 in patients at day 30 after surgery[(53.90±6.53)μg/L]was significantly higher than that at day 1[(43.00±6.37)μg/L,t=-3.804,P<0.01],and that at day 21[(74.00±4.73)μg/L]was significantly higher than that at day 3 after surgery[(57.00±6.87)μg/L,t=-8.046,P<0.01];Transwell showed cell invasion capacity in IGF-1 group(165.67±10.01)was significantly higher than in the control group(50.66±4.04,t=-20.880,P<0.01),the miR-145 expression in IGF-1 group(0.224±0.019)was significantly lower than in the control group(1.000±0.042,t=20.074,P<0.01);the Zeb2 expression in IGF-1 group(0.91±0.25)was significantly higher than in the control group(0.09±0.72,t=-53.959,P<0.01);the expression of Zeb2 protein and cell invasion in the miR-145 inhibitor group(1.33±0.38,259.00±14.10)were significantly higher than in miR-145 mimics group(0.26±0.19,68.00±4.58,P<0.01).Conclusion The increased expression of IGF-1 after hepatectomy may affect the recurrence and metastasis of tumors through the regulation of miR-145.