天津医药

天津医药 ›› 2022, Vol. 50 ›› Issue (3): 241-247.doi: 10.11958/20211157

• 细胞与分子生物学 • 上一篇    下一篇

LincRNA-P21通过靶向miR-17-3p对三阴性乳腺癌细胞迁移和侵袭能力的影响

敖翔，梁红玲，詹勇涛，姜明，夏浩明   

  1. 广州医科大学附属肿瘤医院胸部（乳腺）肿瘤外科（邮编510095）
  • 收稿日期:2021-05-17 修回日期:2021-10-24 出版日期:2022-03-15 发布日期:2022-03-15
  • 基金资助:
    广东省医学科学技术研究基金项目（A2020457）；广州市卫生健康科技项目（20201A010061）

The effect of LincRNA-P21 on metastasis and invasion of triple negative breast cancer cells by targeting miR-17-3p

AO Xiang, LIANG Hongling, ZHAN Yongtao, JIANG Ming, XIA Haoming   

  1. Department of Thoracic (Breast) Oncology, the Affiliated Tumor Hospital of Guangzhou Medical University, Guangzhou 510095, China
  • Received:2021-05-17 Revised:2021-10-24 Published:2022-03-15 Online:2022-03-15

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摘要： 目的　探讨LincRNA-P21通过靶向miR-17-3p对三阴性乳腺癌（TNBC）细胞迁移和侵袭作用的影响。方法　qPCR检测30例TNBC患者肿瘤组织、癌旁正常组织以及6种乳腺细胞（乳腺正常细胞MCF-10A，乳腺癌细胞MDA-MB-231、MDA-MB-435、MDA-MB-468、BT549、T47D）LincRNA-P21和miR-17-3p的表达。取MDA-MB-231细胞，单独过表达LincRNA-P21（实验设对照组、pcDNA3.1空白组、pcDNA-LincRNA-P21组），抑制miR-17-3p表达（实验设对照组、miR-17-3p空白组、miR-17-3p抑制剂组），过表达LincRNA-P21+抑制miR-17-3p表达（实验设对照组、miR-17-3p抑制剂组和pcDNA-LincRNA-P21+miR-17-3p inhibitor组）。CCK-8法检测MDA-MB-231细胞增殖，集落形成实验检测细胞的集落数量，细胞划痕实验检测细胞的迁移能力，Transwell实验检测细胞侵袭能力。Western blot检测E-cadherin和Vimentin蛋白表达。结果　TNBC患者组织中LincRNA-P21的表达明显低于癌旁组织（0.48±0.03 vs. 1.03±0.06，t=11.714，P＜0.01），miR-17-3p的表达显著高于癌旁组织（2.93±0.17 vs. 1.02±0.04，t=15.593，P＜0.01）。乳腺癌细胞系中LincRNA-P21表达量明显低于MCF-10A，miR-17-3p表达量高于MCF-10A（P＜0.01）。单独过表达LincRNA-P21或抑制miR-17-3p后，MDA-MB-231细胞增殖能力、集落形成数量、迁移和侵袭能力均明显下降。过表达LincRNA-P21的同时抑制miR-17-3p，MDA-MB-231细胞增殖能力、集落形成数量、迁移和侵袭能力出现进一步下降，且上调E-cadherin的表达，抑制Vimentin的表达（P＜0.05）。结论　LincRNA-P21通过竞争性结合miR-17-3p来抑制MDA-MB-231细胞增殖、集落形成、迁移和侵袭。

关键词: 三阴性乳腺癌, 细胞运动, RNA干扰, RNA, 长链非编码, LincRNA-P21, miR-17-3p, 细胞侵袭

Abstract: Objective　To investigate the effect of LincRNA-P21 on the migration and invasion of triple negative breast cancer (TNBC) cells by targeting miR-17-3p. Methods　qPCR was used to detect the expression levels of LincRNA-P21 and miR-17-3p in tumor tissues, normal adjacent tissues and 6 kinds of breast cells (normal breast cells MCF-10A, breast cancer cells MDA-MB-231, MDA-MB-435, MDA-MB-468, BT549 and T47D). MDA-MB-231 cells were taken. LincRNA-P21 was overexpressed alone (the control group, the pcDNA3.1 blank group and the pcDNA - LincRNA-P21 group), miR-17-3p expression was inhibited (the control group, the miR-17-3p blank group and the miR-17-3p inhibitor group), and overexpression LincRNA-P21+ inhibited miR-17-3p (the control group, the miR-17-3p inhibitor group and the pcDNA-LincRNA-P21+miR-17-3p inhibitor group). The proliferation of MDA-MB-231 cells was detected by CCK-8 assay. The number of colonies was detected by colony formation assay. The migration ability was detected by cell scratch assay, and the invasion ability was detected by Transwell assay. The protein expressions of E-cadherin and Vimentin were detected by Western blot assay. Results　The expression of LincRNA-P21 was significantly lower in TNBC than that in adjacent tissues (0.48±0.03 vs.1.03±0.06, t=11.714, P＜0.01). The expression of miR-17-3p was significantly higher in TNBC than that in adjacent tissues (2.93±0.17 vs. 1.02±0.04, t=15.593, P＜0.01). The expression levels of LincRNA-P21 mRNA in different breast cancer cell lines were significantly lower than that in MCF-10A (P＜0.01). The expressions of miR-17-3p mRNA in different breast cancer cell lines were significantly higher than that in MCF-10A (P＜0.01). Theoverexpression of LincRNA-P21 inhibited cell proliferation, colony formation and invasion (P＜0.01). The inhibition of miR-17-3p expression could reduce cell proliferation, colony formation and invasion (P＜0.01). Compared with the miR-NC group, the overexpression of LincRNA-P21 and inhibition of miR-17-3p could significantly reduce cell proliferation, colony formation and invasion (P＜0.01), increase the expression of E-cadherin and inhibit the expression of vimentin (P＜0.01). Conclusion　LincRNA-P21 can reduce miR-17-3p expression and inhibit MDA-MB-231 cell proliferation, colony formation, migration and invasion through competitive binding.

Key words: triple negative breast neoplasms, cell movement, RNA interference, RNA, long noncoding, LincRNA-P21, miR-17-3p, cell invasion