中国癌症杂志 ›› 2022, Vol. 32 ›› Issue (9): 818-826.doi: 10.19401/j.cnki.1007-3639.2022.09.009

• 论著 • 上一篇    下一篇

LINC02163靶向miR-338-3p影响乳腺癌细胞增殖、侵袭及迁移

张景臣(), 李新, 李江涛, 李海平, 陈艳丽, 牛冰, 祁川川, 叶贝贝   

  1. 郑州人民医院乳腺科,河南 郑州 450053
  • 收稿日期:2022-01-06 修回日期:2022-03-25 出版日期:2022-09-30 发布日期:2022-10-24
  • 通信作者: 张景臣
  • 基金资助:
    2019年河南省医学科技攻关计划(联合共建)项目(LHGJ20191074)

LINC02163 targeting miR-338-3p affects proliferation, invasion and migration of breast cancer cells

ZHANG Jingchen(), LI Xin, LI Jiangtao, LI Haiping, CHEN Yanli, NIU Bing, QI Chuanchuan, YE Beibei   

  1. Department of Galactophore, People’s Hospital of Zhengzhou, Zhengzhou 450053, Henan Province, China
  • Received:2022-01-06 Revised:2022-03-25 Published:2022-09-30 Online:2022-10-24
  • Contact: ZHANG Jingchen

摘要:

背景与目的: 长链非编码RNA(long non-coding RNA,lncRNA)已被发现在乳腺癌中失调,与肿瘤恶性行为密切相关。本研究旨在探究LINC02163靶向miR-338-3p对乳腺癌细胞增殖、侵袭及迁移的影响。方法: 收集郑州人民医院2020年1月—2021年9月收治的9例乳腺癌患者的乳腺癌组织及距其2 cm外的癌旁组织样本,通过实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR)检测组织样本、人正常乳腺上皮细胞系(MCF-10A)和乳腺癌细胞系(MCF-7、BT-20、MDA-MB-231、T47D)中LINC02163的表达。将MDA-MB-231分为control组、sh-NC组、sh-LINC02163组、sh-LINC02163+inhibitor-NC组和sh-LINC02163+miR-338-3p inhibitor组。采用MTT法、transwell实验及划痕实验分别检测MDA-MB-231细胞活力、侵袭及迁移能力。采用蛋白质印迹法(Western blot)检测MDA-MB-231细胞c-Myc、基质金属蛋白酶(matrix metalloproteinase,MMP)2、MMP9、E-钙粘素(E-cadherin)及N-钙粘素(N-cadherin)蛋白的表达。通过双荧光素酶报告基因实验分析LINC02163与miR-338-3p的靶向关系。采用体内成瘤实验检测BALB/c裸鼠肿瘤体积及重量。采用免疫组织化学法检测裸小鼠移植瘤组织的Ki-67增殖指数。结果: 与癌旁组织相比,LINC02163在乳腺癌组织中的表达显著升高(P<0.05)。与MCF-10A细胞相比,LINC02163在MCF-7、BT-20、MDA-MB-231及T47D细胞中的表达均显著升高,并且其在MDA-MB-231细胞中升高最为显著(P<0.05)。与control组比,sh-LINC02163组LINC02163表达、细胞存活率、侵袭细胞数及迁移率、c-Myc、MMP2、MMP9、N-cadherin表达、肿瘤体积及重量、Ki-67增殖指数显著降低,miR-338-3p、E-cadherin表达显著升高(P<0.05)。与sh-LINC02163组相比,sh-LINC02163+miR-338-3p inhibitor组LINC02163表达无显著变化(P>0.05),细胞存活率、侵袭细胞数及迁移率、c-Myc、MMP2、MMP9、N-cadherin表达、肿瘤体积及重量、Ki-67增殖指数显著增加,miR-338-3p、E-cadherin表达显著降低(P<0.05)。在乳腺癌中LINC02163与miR-338-3p存在潜在的靶向关系。结论: 沉默LINC02163表达通过促进miR-338-3p表达来抑制乳腺癌细胞增殖、侵袭及迁 移。

关键词: LINC02163, miR-338-3p, 乳腺癌, 增殖, 侵袭, 迁移

Abstract:

Background and purpose: Long non-coding RNA (lncRNA) has been found to be dysregulated in breast cancer and is closely related to the malignant behavior of tumors. This study aimed to explore the effects of LINC02163 targeting miR-338-3p on the proliferation, invasion and migration of breast cancer cells. Methods: Breast cancer tissue samples and the adjacent tissue samples (2 cm to cancer tissue) were collected from 9 female breast cancer patients admitted to People’s Hospital of Zhengzhou from January 2020 to September 2021. Real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) was used to detect the expression of LINC02163 in tissue samples, human normal breast epithelial cell line (MCF-10A) and breast cancer cell lines (MCF-7, BT-20, MDA-MB-231, T47D). MDA-MB-231 was divided into control group, sh-NC group, sh-LINC02163 group, sh-LINC02163+inhibitor-NC group and sh-LINC02163+miR-338-3p inhibitor group. MTT method, transwell test and scratch test were used to detect the MDA-MB-231 cell viability, invasion and migration ability. Western blot was used to detect the protein expression of c-Myc, matrix metalloproteinase (MMP) 2, MMP9, E-cadherin and N-cadherin in MDA-MB-231 cells. Dual luciferase reporter gene was used to detect the targeting relationship between LINC02163 and miR-338-3p. In vivo tumor formation experiment was used to detect tumor volume and weight in nude mice. Immunohistochemical method was used to detect the Ki-67 proliferation index in tumor tissues of BALB/c nude mice. Results: Compared with adjacent tissues, the expression of LINC02163 in breast cancer tissues was significantly higher (P<0.05). Compared with MCF-10A cells, the expression of LINC02163 in MCF-7, BT-20, MDA-MB-231 and T47D cells was significantly higher, and it increased most significantly in MDA-MB-231 cells (P<0.05). Compared with the control group, the LINC02163 expression, cell survival rate, number of invasive cells, migration rate, expressions of c-Myc, MMP2, MMP9 and N-cadherin, tumor volume and weight, and Ki-67 proliferation index were significantly reduced in the sh-LINC02163 group, while the expressions of miR-338-3p and E-cadherin were significantly increased (P<0.05). Compared with the sh-LINC02163 group, the expression of LINC02163 in the sh-LINC02163+miR-338-3p inhibitor group did not change significantly (P>0.05), the cell survival rate, number of invasive cells, migration rate, expressions of c-Myc, MMP2, MMP9 and N-cadherin, tumor volume and weight, and Ki-67 proliferation index were significantly increased, while the expressions of miR-338-3p and E-cadherin were significantly reduced (P<0.05). LINC02163 had a potential targeting relationship with miR-338-3p in breast cancer. Conclusion: Silencing the expression of LINC02163 can inhibit the proliferation, invasion and migration of breast cancer cells by promoting the expression of miR-338-3p.

Key words: LINC02163, miR-338-3p, Breast cancer, Proliferation, Invasion, Migration

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