干扰circABCC4靶向微小RNA-185-5p抑制胰腺癌细胞发生发展  

Interfering with circABCC4 targeting microRNA-185-5p to inhibit the development of pancreatic cancer cells

作　　者：于龙 张少康 李超博 高志强[1] Yu Long;Zhang Shaokang;Li Chaobo;Gao Zhiqiang(Department of Hepatic and Biliary Pancreatic Surgery,Henan Key Laboratory of Digestive Organ Transplantation,the First Affiliated Hospitat of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院肝胆胰外科,河南省消化器官移植重点实验室,郑州450052

出　　处：《中华实验外科杂志》2024年第6期1213-1217,共5页Chinese Journal of Experimental Surgery

基　　金：河南省医学科技攻关计划项目(LHGJ20220706)。

摘　　要：目的探讨circABCC4对胰腺癌细胞发生发展的影响及分子机制。方法收集2020年10月至2023年10月郑州大学第一附属医院收治的39例胰腺癌患者的癌组织及癌旁组织,用实时荧光定量聚合酶链反应(RT-qPCR)检测circABCC4和微小RNA(miR)-185-5p的表达水平;双荧光素酶报告实验检测circABCC4和miR-185-5p的靶向关系。将胰腺癌细胞SW1990分为si-circABCC4组、si-NC组、miR-185-5p mimic组、miR-NC组、si-circABCC4+miR-185-5p inhibitor组、si-circABCC4+抗miR-NC组;四甲基偶氮唑盐比色法(MTT)检测细胞增殖抑制率;克隆形成实验检测细胞克隆形成数;流式细胞术检测细胞凋亡率;蛋白质印迹法检测蛋白表达;Transwell检测细胞迁移和侵袭数。两组比较行t检验,多组间比较采用单因素方差分析,组间两两比较采用LSD-t检验。结果胰腺癌组织中circABCC4表达水平高于癌旁组织(3.13±0.29比1.03±0.11,t=42.283,P<0.05),miR-185-5p表达水平低于癌旁组织(0.45±0.08比1.09±0.17,t=21.273,P<0.05)。circABCC4靶向结合并负调控miR-185-5p表达(P<0.05)。si-circABCC4组SW1990细胞抑制率[(47.13±2.23)%比0,F=2376.899,P<0.05]、凋亡率[(18.94±1.08)%比(7.33±0.72)%,F=409.972,P<0.05和B细胞淋巴瘤/白血病-2相关的x蛋白(bax)表达水平(0.55±0.04比0.21±0.03,F=310.551,P<0.05)]高于si-NC组,克隆形成数[(67.44±3.44)个比(112.00±5.94)个,F=296.029,P<0.05]、B细胞淋巴瘤/白血病-2(bcl-2)表达水平(0.27±0.03比0.67±0.05,F=236.571,P<0.05)低于si-NC组。miR-185-5p mimic组SW1990细胞抑制率[(55.52±2.30)%比(0.02±0.01)%,F=2376.899,P<0.05]、凋亡率[(22.59±1.45)%比(7.11±0.55)%,F=409.972,P<0.05]和bax表达水平(0.65±0.04比0.20±0.02,F=310.551,P<0.05)高于miR-NC组,克隆形成数[(54.11±2.23)个比(112.11±6.15)个,F=296.029,P<0.05]、bcl-2表达水平(0.17±0.02比0.67±0.07,F=236.571,P<0.05)低于miR-NC组;si-circABCC4+miR-185-5p inhibitor组SW1990细胞抑制率[(19.13±0.99)%比(47.09±1.73)%,F=2376.899,P<0.05]、凋亡率[(Objective To explore the influence of circABCC4 on the occurrence and development of pancreatic cancer cells and its molecular mechanism.Methods Cancer tissues and adjacent tissues were collected from 39 patients with pancreatic cancer admitted to our hospital from October 2020 to October 2023.Real-time fluorescent quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression levels of circABCC4 and miR-185-5p.The dual luciferase reporter experiment was used to detect the targeting relationship between circABCC4 and miR-185-5p.Pancreatic cancer cells SW1990 were divided into si-circABCC4 group,si-NC group,miR-185-5p mimic group,miR-NC group,si-circABCC4+miR-185-5p inhibitor group,si-circABCC4+anti-miR-NC group.The tetramethylazolium salt colorimetric method(MTT)was used to detect cell proliferation inhibition rate.The clone formation test was used to detect cell clone formation number.The flow cytometry was used to detect cell apoptosis rate.Western blotting was used to detect protein expression.Transwell assay was used to test number of migrating and invasive cells.This study represented the relevant data using mean±standard deviation(±s).The t-test was used for comparing two groups,one-way analysis of variance was used for comparing multiple groups,and LSD-t test was used for pairwise comparisons between groups.P<0.05 was considered to indicate a significant difference.Results Compared to adjacent tissues,the expression of circABCC4 was upregulated(3.13±0.29 vs.1.03±0.11,t=42.283,P<0.05)and the expression of miR-185-5p was downregulated(0.45±0.08 vs.1.09±0.17,t=21.273,P<0.05)in pancreatic cancer tissues.CircABCC4 targeted and negatively regulated miR-185-5p expression(P<0.05).Compared to the si-NC group,SW1990 cells’inhibition rate[(47.13±2.23)%vs.0,F=2376.899,P<0.05],apoptosis rate[(18.94±1.08)%vs.(7.33±0.72)%,F=409.972,P<0.05],and B cell lymphoma/leukemia-2-associated x protein(bax)expression level(0.55±0.04 vs.0.21±0.03,F=310.551,P<0.05)were elevated in the si-circABCC4 group,

关 键 词：胰腺癌 微小RNA 增殖 凋亡 迁移 侵袭 

分 类 号：R735.9[医药卫生—肿瘤]