microRNA-15b通过调节MET-PI3K-Akt通路对肺癌模型鼠免疫功能及高凝状态的影响研究

Effect of microRNA-15b on immune function and hypercoagulable state of lung cancer model mice by regulating the MET-PI3K-Akt pathway

目的探究微小RNA(microRNA,miR)-15b通过调节MET-PI3K-Akt通路对肺癌模型鼠免疫功能及高凝状态的影响。方法人肺癌细胞A549分为阴性对照(negative control,NC)组、mimic组和inhibitor组,分别转染NC、miR-15b mimic和miR-15b inhibitor,通过皮下注射的方式构建肺癌荷瘤裸鼠模型。TUNEL染色检测凋亡。Western blot检测增殖蛋白CyclinD1、Ki67以及MET-PI3K-Akt通路水平。检测和比较两组的CD4^(+)、CD8^(+)水平与凝血功能。结果与NC组比较,mimic组的miR-15b水平(3.45±0.36)、凋亡指数(17.72±3.57)%、PT、APTT、NO、CD4^(+)(19.83±2.71%)和CD4^(+)/CD8^(+)水平(1.30±0.34)显著升高(P<0.05),CyclinD1和Ki67蛋白水平、全血黏度、ET1、MET、PI3K、Akt水平显著降低(P<0.05)。inhibitor组的miR-15b水平(0.52±0.07)、凋亡指数(1.35±0.41)%、PT、APTT、NO、CD4^(+)(19.83±2.71%)和CD4^(+)/CD8^(+)水平(1.30±0.34)降低(P<0.05),CyclinD1和Ki67蛋白水平、全血黏度、ET1、MET、PI3K、Akt水平升高(P<0.05)。结论在肺癌裸鼠模型中,miR-15b可以诱导肺癌细胞的凋亡和抑制增殖,并促进免疫功能改善高凝状态。

Objective To explore the effect of microRNA(miR)-15b on immune function and hypercoagulability of lung cancer model rats by regulating the MET-PI3K-Akt pathway.Methods Human lung cancer cells A549 were divided into the negative control(NC)group,the mimic group and the inhibitor group,and they were transfected with NC,miR-15b mimic and miR-15b inhibitor,respectively.A nude mouse model of lung cancer was constructed by subcutaneous injection.TUNEL staining was used to detect apoptosis.Western blot was used to detect the levels of the proliferating proteins CyclinD1,Ki67 and MET-PI3K-Akt pathway.The levels of CD4+and CD8+in the two groups were detected and compared for coagulation function.Results Compared with the NC group,the miR-15b level(3.45±0.36),apoptosis index(17.72±3.57)%,PT,APTT,NO,CD4+(19.83±2.71%)and CD4+/CD8+level(1.30±0.34)increased significantly in the mimic group(P<0.05),and CyclinD1 and Ki67 protein levels,whole blood viscosity,ET1,MET,PI3K,and Akt levels decreased significantly(P<0.05).The miR-15b level(0.52±0.07),apoptosis index(1.35±0.41)%,PT,APTT,NO,CD4^(+)(19.83±2.71%)and CD4^(+)/CD8^(+) levels(1.30±0.34)in the inhibitor group decreased(P<0.05),and CyclinD1 and Ki67 protein levels,whole blood viscosity,ET1,MET,PI3K,and Akt levels increased(P<0.05).Conclusion In the nude mouse model of lung cancer,miR-15b can induce apoptosis and proliferation of lung cancer cells,and promote immune function to improve hypercoagulability.