circ-MAT2B调节前列腺癌细胞DU145生物学行为的分子机制  

Molecular mechanism of circ-MAT2B regulating the biological behavior of prostate cancer cell DU145

作　　者：杨君[1] 江波涛[1] 孔东波[1] 汪洋[1] YANG Jun;JIANG Bo-tao;KONG Dong-bo;WANG Yang(Department of Urology,Xianning Central Hospital,The First Affiliated Hospital of Hubei University of Science and Technology,Xianning 437100,Hubei,China)

机构地区：[1]咸宁市中心医院湖北科技学院附属第一医院泌尿外科,咸宁437100

出　　处：《医学研究生学报》2022年第8期833-840,共8页Journal of Medical Postgraduates

基　　金：湖北省卫生健康委面上项目(WJ2021M091)。

摘　　要：目的 circ-MAT2B在前列腺癌中的表达及其可能作用机制尚不清楚。文中旨在探讨circ-MAT2B调控miR-491-5p/PEG10轴对前列腺癌细胞DU145增殖、迁移、凋亡的影响。方法 收集2019年1月至2020年1月咸宁市中心医院泌尿外科接受根治性切除手术的37份前列腺癌组织和正常邻近组织样本(癌旁组织)。根据待转染物不同将DU145细胞分为si-NC组、si-circ-MAT2B组、miR-NC组、miR-491-5p mimic组、si-circ-MAT2B+miR-491-5p inhibitor组,未转染DU145细胞记为对照组。RT-qPCR检测前列腺癌组织和对应癌旁组织中circ-MAT2B和miR-491-5p相对水平;平板克隆实验、CCK-8实验检测DU145增殖能力;划痕愈合实验和Transwell实验检测DU145迁移能力;流式细胞术检测细胞凋亡率;蛋白质印记法分析PEG10蛋白表达;双荧光素酶报告实验分析miR-491-5p与circ-MAT2B或PEG10的相互作用。结果 前列腺癌组织中circ-MAT2B相对水平(4.04±0.33)较癌旁组织(1.04±0.13)显著升高(P<0.05),miR-491-5p相对水平(0.25±0.05)较癌旁组织(1.00±0.10)显著降低(P<0.05)。与对照组、si-NC组比较,si-circ-MAT2B2组DU145细胞miR-491-5p表达、凋亡率显著升高(P<0.05),PEG10蛋白表达、细胞存活率、克隆形成数、迁移距离、侵袭数显著降低(P<0.05)。与miR-NC组比较,miR-491-5p mimic组DU145细胞凋亡率显著升高(P<0.05),PEG10蛋白表达、细胞存活率、克隆形成数、迁移距离、侵袭数显著降低(P<0.05)。与si-circ-MAT2B2组比较,si-circ-MAT2B+miR-491-5p inhibitor组DU145细胞凋亡率显著降低(P<0.05),PEG10蛋白表达、细胞存活率、克隆形成数、迁移距离、侵袭数显著升高(P<0.05)。miR-491-5p分别与circ-MAT2B、PEG10直接结合。结论 干扰circ-MAT2B可诱导前列腺癌细胞DU145凋亡,抑制其增殖和迁移,其抗肿瘤机制可能与上调miR-491-5p/PEG10轴有关。Objective The expression of circ-MAT2 B in prostate cancer and its possible mechanism of action were not yet known. This study aimed to investigate the effect of circ-MAT2 B regulating miR-491-5 p/PEG10 axis on the proliferation, migration and apoptosis of prostate cancer cells DU145. Methods Thirty-seven samples of prostate cancer tissues and normal adjacent tissues(para-cancer tissues) from patients undergoing radical prostatectomy in the Department of Urology, Xianning Central Hospital from January 2019 to January 2020 were collected. DU145 cells were divided into four groups according to different transfection materials: si-NC group, si-circ-MAT2 B group, miR-NC Group, miR-491-5 p mimic group, si-circ-MAT2 B + Mir-491-5 P inhibitor group. DU145 cells without transfection were recorded as the control group. The relative levels of circ-MAT2 B and miR-491-5 p in prostate cancer tissues and corresponding adjacent tissues were calculated using RT-qPCR. Plate cloning assays and CCK-8 methods were used to detect DU145 proliferation ability;scratch healing assays and Transwell assays were conducted to explore DU145 migration ability;flow cytometry was used to exam cell apoptosis rate;Western blot was applied to analyze PEG10 protein expression. The interaction between miR-491-5 p and circ-MAT2 B or PEG10 was analyzed by dual luciferase report assay. Results The relative level of circ-MAT2 B(4.04±0.33) was significantly higher than that of the adjacent tissues(1.04±0.13)(P<0.05), and the relative level of miR-491-5 p(0.25±0.05) was significantly lower than that of the adjacent tissues(1.00±0.10)(P<0.05). Compared with the control group and si-NC group, the miR-491-5 p expression, apoptosis rate of DU145 cells in the si-circ-MAT2 B group notably increased(P<0.05), while PEG10 protein expression, cell survival rate, clonal formation numbers, migration distance and invasion numbers were notably reduced(P<0.05). Compared with the miR-NC group, the apoptosis rate of DU145 cells in the miR-491-5 p mimic group notably in

关 键 词：circ-MAT2B 前列腺癌 细胞增殖 凋亡 迁移 miR-491-5p PEG10 

分 类 号：R737.25[医药卫生—肿瘤]