miR-106a靶向调控TLR4对A549细胞增殖的作用机制  

Mechanism of miR-106a targeting regulation of TLR4 on A549 cell proliferation

作　　者：高少林[1] 王晓丹 朱海勇 马政 芦旭超 GAO Shao-lin;WANG Xiao-dan;ZHU Hai-yong;MA Zheng;LU Xu-chao(Department of Thoracic Surgery,Second Hospital of Hebei Medical University,Shijiazhuang 050000,China)

机构地区：[1]河北医科大学第二医院胸外科,河北石家庄050000

出　　处：《生物技术》2023年第5期581-585,557,共6页Biotechnology

摘　　要：[目的]验证miR-106a对Toll受体4(TLR4)的靶向性,探讨其对肺癌细胞系A549增殖的调控作用。[方法]采用TCGA数据库和双荧光素酶实验分析miR-106a与TLR4相关性,将A549细胞分为NC组、inhibitor组、inhibitor+TLR4组和TLR4组,RT-PCR检测细胞miR-106a、TLR4 mRNA表达,CCK8检测细胞增殖水平,流式细胞仪检测细胞凋亡,Western Blot检测TLR4、细胞周期蛋白(Cyclin)、增殖细胞核抗原(PCNA)、凋亡蛋白(Bax)、抗凋亡蛋白(Bcl-2)蛋白表达表达。[结果]双荧光素酶实验显示过表达miR-106a能提高TLR4蛋白表达(1.00±0.12 vs 1.39±0.16,P<0.05);与NC组比较,inhibitor组miR-106a、TLR4 mRNA及蛋白表达、细胞增殖、Bcl-2、PCNA、Cyclin蛋白表达均降低,凋亡率、Bax表达升高(P<0.05),TLR4组miR-106a、TLR4 mRNA及蛋白表达、细胞增殖、Bcl-2、PCNA、Cyclin蛋白表达升高,凋亡率、Bax表达降低(P<0.05);与inhibitor组比较,inhibitor+TLR4组miR-106a、TLR4 mRNA及蛋白表达、细胞增殖、Bcl-2、PCNA、Cyclin蛋白表达升高(P<0.05),凋亡率、Bax表达降低(P<0.05)。[结论]miR-106a可通过靶向促进TLR4表达来促进肺癌A549细胞增殖,抑制其凋亡,从而发挥促癌作用。[Objective]To verify the targeting of miR-106a to Toll receptor 4(TLR4)and investigate its regulation on the proliferation of A549 cell.[Method]The correlation between miR-106a and TLR4 was analyzed by TCGA database and dual-luciferase assay.A549 cells were divided into a NC group,an inhibitor group and an inhibitor+TLR4 group.The expression of miR-106a and TLR4 mRNA was detected by RT-PCR,the proliferation level was detected by CCK8,the apoptosis was detected by flow cytometry,and the expression of TLR4,cyclin,proliferating cell nuclear antigen(PCNA),apoptotic protein(Bax)and anti-apoptotic protein(Bcl-2)protein was detected by Western Blot.[Result]Dual-luciferase assay showed that overexpression of miR-106a could increase TLR4 protein expression(P<0.05).Compared with the NC group,miR-106a and TLR4 mRNA and protein expression,cell proliferation,Bcl-2,PCNA,and cyclin protein expression were decreased,while apoptosis rate and Bax expression were increased in the inhibitor group(P<0.05);miR-106a and TLR4 mRNA and protein expression,cell proliferation,Bcl-2,PCNA,and cyclin protein expression were increased,and apoptosis rate and Bax expression were decreased in the TLR4 group(P<0.05).Compared with the inhibitor group,miR-106a and TLR4 mRNA and protein expression,cell proliferation,Bcl-2,PCNA,and cyclin protein expression were increased(P<0.05),and apoptosis rate and Bax expression were decreased in the inhibitor+TLR4 group(P<0.05).[Conclusion]miR-106a can promote the proliferation and inhibit the apoptosis of lung cancer A549 cells by targeting and promoting the expression of TLR4,thus exerting a cancer-promoting effect.

关 键 词：miR-106a 肺癌 Toll受体4 细胞增殖 

分 类 号：R734.2[医药卫生—肿瘤]