摘要
目的分析长链非编码RNA(lncRNA)RP11-686D22.10表达水平与乳腺癌患者生存期的关系,探讨过表达RP11-686D22.10对乳腺癌细胞系增殖和侵袭的影响及其分子机制。方法GEPIA数据库分析RP11-686D22.10表达水平与乳腺癌患者生存期的关系。RT-qPCR检测乳腺上皮细胞系MCF-10A和乳腺癌细胞系MCF7、SKBR3、HCC1937、BT549和MB-MDA-468中RP11-686D22.10的表达量。选择RP11-686D22.10表达水平最低的SKBR3细胞,分别转染对照质粒(NC组)和RP11-686D22.10过表达质粒(RP11-686D22.10组)。CCK-8法和Transwell小室法分别检测乳腺癌细胞增殖及侵袭。Linc2Go数据库预测RP11-686D22.10的靶向微小RNA(miRNA),双荧光素酶报告基因实验检测RP11-686D22.10和miR-454-3p的靶向关系。RT-qPCR检测miR-454-3p表达量。Western blot检测Wnt/β-catenin信号通路蛋白质表达量。结果与RP11-686D22.10低表达乳腺癌患者相比,RP11-686D22.10高表达患者的总生存期较长(P<0.01),并且无病生存期较长(P<0.01)。MCF7、SKBR3、HCC1937、BT549和MB-MDA-468乳腺癌细胞中RP11-686D22.10表达量低于MCF-10A细胞(P<0.05),SKBR3细胞中RP11-686D22.10表达最低(P<0.01)。与NC组比较,RP11-686D22.10组SKBR3细胞增殖能力降低(P<0.05),并且细胞侵袭能力降低(P<0.01)。RP11-686D22.10能够靶向结合miR-454-3p(P<0.01)。与NC组比较,RP11-686D22.10组miR-454-3p表达量降低(P<0.01),Wnt/β-catenin信号通路蛋白CCND1、β-catenin、JUN、AXIN2和MMP2表达量降低。结论RP11-686D22.10表达水平与乳腺癌患者生存期密切相关,过表达RP11-686D22.10通过靶向调控miR-454-3p抑制乳腺癌细胞系的增殖及侵袭。
Objective To dientify the correlation between the expression of long-chain non-coding RNA(lncRNA)RP11-686D22.10 and the survival of breast cancer patients,and to explore the effect of over-expression of RP11-686D22.10 on the proliferation and invasion of breast cancer cell lines.Methods The relationship between the expression level of RP11-686D22.10 and the survival time of breast cancer patients was analyzed with GEPIA database.The expression of RP11-686D22.10 in mammary epithelial cell line MCF-10A and breast cancer cell lines MCF7,SKBR3,HCC1937,BT549 and MB-MDA-468 was detected by real-time quantitative polymerase chain reaction(RT-qPCR).SKBR3 cell with the least expression of RP11-686D22.10 was selected and transfected with control plasmid(NC group)and RP11-686D22.10 over-expression plasmid(RP11-686D22.10 group).The proliferation and invasion of breast cancer cells were detected by CCK-8 and Transwell assays,respectively.The Linc2Go database predicted the targeting microRNA(miRNA)of RP11-686D22.10,and the dual-luciferase reporter gene assay was used to target at relationship between RP11-686D22.10 and miR-454-3p.The expression of miR-454-3p was detected by RT-qPCR.Western blot was used to detect the protein expression of Wnt/β-catenin signaling pathway.Results Compared with breast cancer patients with low expression of RP11-686D22.10,patients with high expression of RP11-686D22.10 had longer overall survival(P<0.01)and longer disease-free survival(P<0.01).The expression of RP11-686D22.10 in MCF7,SKBR3,HCC1937,BT549,and MB-MDA-468 cells was lower than that in MCF-10A cell(P<0.05).The expression of RP11-686D22.10 in SKBR3 cells was the lowest(P<0.01).Compared with the NC group,the proliferation ability of SKBR3 cells in the RP11-686D22.10 group was decreased(P<0.05),and the cell invasion ability was decreased(P<0.01).RP11-686D22.10 could target and bind miR-454-3p(P<0.01).Compared with the NC group,the expression of miR-454-3p in the RP11-686D22.10 group was decreased(P<0.01),and the expressions of Wnt/β-catenin signaling pathway proteins CCND1,β-catenin,JUN,AXIN2,and MMP2 was decreased.Conclusions The expression level of RP11-686D22.10 is closely related to the survival of breast cancer patients.Over-expression of RP11-686D22.10 inhibits the proliferation and invasion of breast cancer cell lines by targeting at miR-454-3p.
作者
唐一君
王文龙
李江丽
杨欢
孙静
TANG Yijun;WANG Wenlong;LI Jiangli;YANG Huan;SUN Jing(Department of the Fifth Ward of Medical Oncology;Department of the Fifth Ward of Surgery,Anyang Cancer Hospital Affiliated to Henan University of Science and Technology,Anyang 455000;Department of Thyroid and Breast Surgery,the First Affiliated Hospital of Soochow University,Suzhou 215006,China)
出处
《基础医学与临床》
2023年第7期1023-1029,共7页
Basic and Clinical Medicine
基金
国家自然科学基金(81702078)。
