lncRNA SNHG 1对乳腺癌细胞迁移、侵袭和上皮-间质转化的影响及其机制  

EFFECT OF LONG NON-CODING RNA SMALL NUCLEOLAR RNA HOST GENE 1 ON MIGRATION,INVASION,AND EPITHELIAL-MESENCHYMAL TRANSITION OF BREAST CANCER CELLS AND ITS MECHANISM

作　　者：邓林 吴清兰 宋军莹 高笑 肖欢欢 张丽[2] 曹奕[3] 侯琳[1] DENG Lin;WU Qinglan;SONG Junying;GAO Xiao;XIAO Huanhuan;ZHANG Li;CAO Yi;HOU Lin(Department of Biochemistry and Molecular Bio-logy,School of Basic Medicine,Qingdao University,Qingdao 266071,China)

机构地区：[1]青岛大学基础医学院生物化学与分子生物学系,山东青岛266071 [2]青岛大学药学院,山东青岛266071 [3]青岛大学生物医学实验中心,山东青岛266071

出　　处：《精准医学杂志》2023年第2期159-163,168,共6页Journal of Precision Medicine

基　　金：国家自然科学基金资助项目(81472542);山东省重点研发项目(2019GSF107025);山东省医药卫生科技发展计划项目(202102021141);山东省中医药科技项目(2021Z196);山东省科技卫生发展计划项目(202102021147)。

摘　　要：目的探讨长链非编码RNA(lncRNA)核仁小分子RNA宿主基因1(SNHG 1)对乳腺癌细胞迁移、侵袭和上皮-间质转化(EMT)的影响及其作用机制。方法收集20例乳腺癌患者的癌组织与癌旁正常组织,培养人正常乳腺上皮细胞MCF-10A及乳腺癌细胞MCF-7、BT-549、MDA-MB-231和MDA-MB-468,采用实时荧光定量PCR(RT-qPCR)方法检测组织和细胞中SNHG 1与miR-641表达量。以BT-549细胞为研究对象,分为A~D组,分别转染si-NC(A组)、si-SNHG1(B组)、si-SNHG1+anti-miR-641-NC(C组)、si-SNHG1+anti-miR-641(D组),采用RT-qPCR检测细胞中SNHG 1和miR-641表达量,采用Transwell实验检测细胞侵袭和迁移能力,用Western blot检测细胞EMT相关蛋白E-cadherin、N-cadherin和Vimentin表达量。将BT-549细胞分为E~H组,分别共转染SNHG1-WT与miR-641 mimics(E组)、SNHG1-WT与miR-NC(F组)、SNHG1-MUT与miR-641 mimics(G组)以及SNHG1-MUT与miR-NC(H组),采用双荧光素酶报告基因实验验证SNHG 1和miR-641间是否存在互作。结果SNHG 1在乳腺癌组织中高表达(t=4.81,P<0.05),miR-641在乳腺癌组织中低表达(t=7.96,P<0.05)。与正常乳腺上皮细胞MCF-10A相比,4种乳腺癌细胞中SNHG 1表达明显上调(t=8.11~10.79,P<0.05),而miR-641表达明显降低(t=9.16~11.17,P<0.05)。相较于A组,B组BT-549细胞的迁移及侵袭能力降低,N-cadherin和Vimentin蛋白表达量降低,E-cadherin蛋白和miR-641表达升高(t=3.95~19.45,P<0.05)。相较于C组,D组BT-549细胞迁移、侵袭能力增强,N-cadherin和Vimentin蛋白表达升高,E-cadherin蛋白表达降低(t=3.59~13.83,P<0.05)。双荧光素酶报告基因实验结果显示,E组荧光素酶活性低于F组(t=8.37,P<0.05),G组与H组荧光素酶活性比较无统计学差异(P>0.05)。结论SNHG 1在乳腺癌组织和细胞中高表达,下调其表达可通过靶向miR-641抑制乳腺癌细胞迁移、侵袭及EMT过程。Objective To investigate the effect of the long non-coding RNA(lncRNA)small nucleolar RNA host gene 1(SNHG 1)on the migration,invasion,and epithelial-mesenchymal transition(EMT)of breast cancer cells and its mechanism.Methods Breast cancer tissue samples and adjacent normal tissue samples were collected from 20 patients with breast cancer,and normal human breast epithelial cell MCF-10A and breast cancer cell lines MCF-7,BT-549,MDA-MB-231,and MDA-MB-468 were cultured.RT-qPCR was used to measure the expression levels of SNHG 1 and miR-641 in tissue samples and cells.BT-549 cells were divided into groups A to D and were transfected with si-NC(group A),si-SNHG1(group B),si-SNHG1+anti-miR-641-NC(group C),and si-SNHG1+anti-miR-641(group D),respectively.RT-qPCR was used to measure the expression levels of SNHG 1 and miR-641 in cells;Transwell assay was used to observe cell invasion and migration abilities;Western blot was used to measure the expression levels of the EMT-related proteins E-cadherin,N-cadherin,and Vimentin.BT-549 cells were divided into groups E to H and were co-transfected with SNHG1-WT and miR-641 mimics(group E),SNHG1-WT and miR-NC(group F),SNHG1-MUT and miR-641 mimics(group G),and SNHG1-MUT and miR-NC(group H),respectively,and dual-luciferase reporter assay was used to determine the interaction between SNHG 1 and miR-641.Results SNHG 1 was highly expressed in breast cancer tissue(t=4.81,P<0.05),while miR-641 was lowly expressed in breast cancer tissue(t=7.96,P<0.05).Compared with normal human breast epithelial cell line MCF-10A,the four breast cancer cell lines had a significantly upregulated expression of SNHG1(t=8.11-10.79,P<0.05)and a significant reduction in the expression of miR-641(t=9.16-11.17,P<0.05).Compared with group A,group B had significant reductions in the migration and invasion abilities of BT-549 cells and the protein expression le-vels of N-cadherin and Vimentin,as well as significant increases in the expression levels of N-cadherin and miR-641(t=3.95-19.45,P<0.05).Compared with grou

关 键 词：乳腺肿瘤 长链非编码RNA 微RNAS 细胞运动 肿瘤浸润 上皮-间质转化 

分 类 号：R737.9[医药卫生—肿瘤]