基于miR-106b-5p调控CCND1基因研究幽门螺杆菌促进食管癌细胞增殖的功能机制

Study on the functional mechanism of Helicobacter pylori on promoting the proliferation of esophageal cancer cells based on the regulation of CCND1 gene by miR-106b-5p

目的研究幽门螺杆菌(Hp)感染通过miR-106b-5p/细胞周期蛋白D1(CCND1)途径促进食管癌细胞增殖的作用机制。方法收集手术切除的食管癌组织46份,检测Hp感染情况及miR-106b-5p、CCND1的表达水平。培养食管癌TE-1细胞株,分别给予不处理、Hp感染、转染阴性对照(NC)序列、转染miR-106b-5p模拟物、转染NC序列同时加入Hp菌液、转染miR-106b-5p模拟物同时加入Hp菌液处理,分别作为对照组、Hp组、miR-NC组、miR-106b-5p组、miR-NC+Hp组、miR-106b-5p+Hp组,检测细胞增殖情况、细胞周期及miR-106b-5p、CCND1的表达水平。结果Hp阳性的食管癌组织中miR-106b-5p的表达水平低于Hp阴性的食管癌组织,CCND1的表达水平高于Hp阴性的食管癌组织,差异有统计学意义(P<0.05)。食管癌组织中miR-106b-5p与CCND1的表达水平呈负相关(r=-0.414,P=0.004);Hp组TE-1细胞的A_(490)水平、细胞周期S期及G2/M期比例、CCND1的表达水平高于对照组,G0/G1期比例及miR-106b-5p的表达水平低于对照组,差异有统计学意义(P<0.05);miR-106b-5p组TE-1细胞中野生型CCND1报告基因的荧光值低于对照组,差异有统计学意义(P<0.05);miR-106b-5p+Hp组TE-1细胞的A_(490)水平、细胞周期S期及G2/M期比例、CCND1的表达水平低于miR-NC+Hp组,G0/G1期比例及miR-106b-5p的表达水平高于miR-NC+Hp组,差异有统计学意义(P<0.05)。结论Hp感染食管癌细胞促进细胞增殖、加速细胞周期,这一作用与调控miR-106b-5p/CCND1途径有关。

Objective To study the role and mechanism of Helicobacter pylori infection in promoting the proliferation of esophageal cancer cells through miR-106b-5p/cyclin D1(CCND1)pathway.Methods A total of 46 patients resected esophageal cancer tissues were collected,and the Hp infection,the expression levels of miR-106b-5p and CCND1 were detected.The esophageal cancer TE-1 cell was cultured and given no processing,Hp infection,transfected with negative control(NC)sequence,transfected miR-106b-5p mimic,transfected NC sequence and added Hp,transfected miR-106b-5p mimic and added Hp,used as control group,Hp group,miR-NC group,miR-106b-5p group,miR-NC+Hp group,miR-106b-5p+Hp group.The cell proliferation,cell cycle and the expression levels of miR-106b-5p and CCND1 were detected.Results The expression level of miR-106b-5p in Hp positive esophageal cancer tissues was lower than that in Hp negative esophageal cancer tissues,but the expression level of CCND1 was higher than that in Hp negative esophageal cancer tissues(P<0.05).The expression level of miR-106b-5p correlated negatively with the expression level of CCND1(r=-0.414,P=0.004).The cell A_(490) level,cell cycle S phase,G2/M phase ratio and CCND1 expression level of TE-1 cells in Hp group were higher than those in control group,while the G0/G1 phase ratio and miR-106b-5p expression level were lower than those in control group(P<0.05).The fluorescence value of wild-type CCND1 reporter gene in TE-1 cells of miR-106b-5p group was lower than that in control group(P<0.05).The cell A_(490)level,cell cycle S phase and G2/M phase ratio,CCND1 expression level of TE-1 cells in miR-106b-5p+Hp group were lower than those in miR-NC+Hp group,while the G0/G1 phase ratio and miR-106b-5p expression level were higher than those in miR-NC+Hp group(P<0.05).Conclusion Hp infection of esophageal cancer cells promotes cell proliferation and accelerates cell cycle,which relates to the regulation of miR-106b-5p/CCND1 pathway.