基础医学与临床 ›› 2023, Vol. 43 ›› Issue (7): 1023-1029.doi: 10.16352/j.issn.1001-6325.2023.07.1023

• 研究论文 •    下一篇

LncRNA RP11-686D22.10与乳腺癌患者生存期相关并影响乳腺癌细胞系的增殖和侵袭

唐一君1, 王文龙2,*, 李江丽1, 杨欢3, 孙静1   

  1. 1.河南科技大学附属安阳市肿瘤医院 肿瘤内科五病区, 河南 安阳 455000;
    2.河南科技大学附属安阳市肿瘤医院 外科五病区, 河南 安阳 455000;
    3.苏州大学附属第一医院 甲乳外科, 江苏 苏州 215006
  • 收稿日期:2022-07-07 修回日期:2022-11-13 出版日期:2023-07-05 发布日期:2023-07-05
  • 通讯作者: *wwlnanyang@163.com
  • 基金资助:
    国家自然科学基金(81702078)

LncRNA RP11-686D22.10 is related with the survival of breast cancer patients, affects the proliferation, invasion of breast cancer cell lines

TANG Yijun1, WANG Wenlong2,*, LI Jiangli1, YANG Huan3, SUN Jing1   

  1. 1. Department of the Fifth Ward of Medical Oncology, Anyang Cancer Hospital Affiliated to Henan University of Science and Technology, Anyang 455000;
    2. Department of the Fifth Ward of Surgery, Anyang Cancer Hospital Affiliated to Henan University of Science and Technology, Anyang 455000;
    3. Department of Thyroid and Breast Surgery, the First Affiliated Hospital of Soochow University, Suzhou 215006, China
  • Received:2022-07-07 Revised:2022-11-13 Online:2023-07-05 Published:2023-07-05

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。

关键词: 乳腺癌, RP11-686D22.10, miR-454-3p, 增殖, 侵袭

Abstract: 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.

Key words: breast cancer, RP11-686D22.10, miR-454-3p, proliferation, invasion

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