miR-183-5p靶向RBMS1对乳腺癌细胞自噬和迁移的影响  

Effects of miR-183-5p targeting RBMS1 on autophagy and migration of breast cancer cells

作　　者：陈登云[1] 吴一波[1] 黄琨波 张汉辉 张志珊[1] 洪志鹏 CHEN Deng-yun;WU Yi-bo;HUANG Kun-bo;ZHANG Han-hui;ZHANG Zhi-shan;HONG Zhi-peng(Clinical Laboratory and Department,QuanzhouFirst Hospital,Quanzhou 362000,Fujian Province,China;Breast Department,QuanzhouFirst Hospital,Quanzhou 362000,Fujian Province,China)

机构地区：[1]泉州市第一医院检验科,福建泉州362000 [2]泉州市第一医院乳腺科,福建泉州362000

出　　处：《中国临床药理学杂志》2025年第5期671-675,共5页The Chinese Journal of Clinical Pharmacology

基　　金：福建省自然科学基金资助项目(2023J011775)。

摘　　要：目的探讨微小RNA-183-5p(miR-183-5p)靶向RNA结合基序单链相互作用蛋白1(RBMS1)对乳腺癌细胞自噬和迁移的影响及其机制。方法将人乳腺癌细胞MCF-7分为对照组、NC inhibitor组(转染NC inhibitor)、miR-183-5p inhibitor组(转染miR-183-5p inhibitor)、miR-183-5p inhibitor+si-NC组(转染空载体miR-183-5p inhibitor和siNC)、miR-183-5p inhibitor+si-RBMS1组(转染RBMS1敲低质粒si-RBMS1)。蛋白质印迹法检测细胞自噬和磷脂酰肌醇-3-激酶(PI3K)/苏氨酸蛋白激酶B(Akt)/雷帕霉素靶蛋白(mTOR)通路相关蛋白的相对表达水平,划痕实验检测细胞的迁移率。结果对照组、NC inhibitor组、miR-183-5p inhibitor组、miR-183-5p inhibitor+si-NC组和miR-183-5p inhibitor+si-RBMS1组微管相关蛋白轻链3Ⅱ(LC3-Ⅱ)/微管相关蛋白轻链3Ⅰ(LC3-Ⅰ)蛋白的相对表达水平分别为0.29±0.03、0.31±0.03、0.86±0.10、0.84±0.09和0.43±0.05,自噬效应蛋白(Beclin-1)的相对表达水平分别为0.18±0.02、0.20±0.02、0.74±0.08、0.78±0.09和0.35±0.04,自噬蛋白(P62)蛋白的相对表达水平分别为0.93±0.12、0.89±0.10、0.51±0.06、0.54±0.06和0.86±0.10,48 h细胞迁移率分别为(62.87±7.26)%、(59.23±6.89)%、(24.13±3.49)%、(26.14±4.72)%和(40.11±5.71)%,磷酸化-(p-)PI3K/PI3K蛋白的相对表达水平分别为0.67±0.12、0.64±0.10、0.32±0.06、0.35±0.06和0.74±0.09,miR-183-5p inhibitor+si-RBMS1组的上述指标与miR-183-5p inhibitor+si-NC组相比,miR-183-5p inhibitor组的上述指标与NC inhibitor组相比,在统计学上差异均有统计学意义(均P<0.05)。结论抑制miR-183-5p表达能促进MCF-7细胞自噬,抑制细胞迁移,下调RBMS1则有相反结果,其机制与PI3K/Akt/mTOR通路有关。Objective To investigate the effect of microRNA-183-5p(miR-183-5p)targeting RNA-binding motif single-strandedinteracting protein 1(RBMS1)on autophagy and migration of breast cancer cells and its mechanism.Methods Human breast cancer cells MCF-7 were divided into control group,NC inhibitor group(transfected with NC inhibitor)and miR-183-5p inhibitor group(transfected with miR-183-5p inhibitor),miR-183-5p inhibitor+si-NC group(transfected with miR-183-5p inhibitor and empty vector si-NC),miR-183-5p inhibitor+si-RBMS1 group(transfected with miR-183-5p inhibitor and RBMS1 knockdown plasmid si-RBMS1).The expression levels of autophagy and phosphatidylinositol-3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR)pathway-related proteins in each group were detected by Western blot,and the mobility of cells in each group was detected by scratch test.Results Control group,NC inhibitor group,miR-183-5p inhibitor group,miR-183-5p inhibitor+si-NC group and miR-183-5p inhibitor+si-RBMS1 group the relative expression levels of light chain 3Ⅱ(LC3-Ⅱ)/light chain 3Ⅰ(LC3-Ⅰ)protein were 0.29±0.03,0.31±0.03,0.86±0.10,0.84±0.09and 0.43±0.05,respectively;the relative expression levels of Beclin-1 protein were 0.18±0.02,0.20±0.02,0.74±0.08,0.78±0.09 and 0.35±0.04,respectively;the relative expression levels of sequestosome-1(P62)protein were 0.93±0.12,0.89±0.10,0.51±0.06,0.54±0.06 and 0.86±0.10,respectively;the 48 h cell mobility was(62.87±7.26)%,(59.23±6.89)%,(24.13±3.49)%,(26.14±4.72)%and(40.11±5.71)%,respectively;the relative expression levels of phospho-(p-)PI3K/PI3K protein were0.67±0.12,0.64±0.10,0.32±0.06,0.35±0.06 and 0.74±0.09,respectively.The above indexes of miR-183-5p inhibitor+si-RBMS1 group were compared with those of miR-183-5p inhibitor+si-NC group,and those of miR-183-5p inhibitor group were compared with those of NC inhibitor group,and the differences were statistically significant(allP<0.05).Conclusion Inhibition ofmiR-183-5pexpression can promote autophagy and inhib

关 键 词：微小RNA-183-5p 乳腺癌 细胞自噬 细胞迁移 RNA结合基序单链相互作用蛋白1 磷脂酰肌醇-3-激酶/苏氨酸蛋白激酶B/雷帕霉素靶蛋白通路 

分 类 号：R97[医药卫生—药品]