实用医学杂志 ›› 2020, Vol. 36 ›› Issue (24): 3338-3343.doi: 10.3969/j.issn.1006⁃5725.2020.24.007

• 基础研究 • 上一篇    下一篇

miR⁃17⁃5p通过下调肌球蛋白调节轻链相互作用蛋白促进结直肠癌细胞增殖和转移的机制研究

凌旭坤, 谢文鸿 ,张喆, 胡琛   

  1. 惠州市中心人民医院胃肠外科(广东惠州 516200)
  • 出版日期:2020-12-25 发布日期:2021-01-19
  • 通讯作者: 凌旭坤 E⁃mail:lxk6534@163.com
  • 基金资助:
    广东省医学科研基金立项项目(编号:B2018263)

Mechanism of miR⁃17⁃5p in promoting proliferation and metastasis of colorectal cancer cells by down⁃regu⁃lating MYLIP

LING Xukun,XIE Wenhong,ZHANG Zhe,HU Chen
  

  1. Department of Gastrointestinal Surgery,Hui⁃zhou Central People′s Hospital,Huizhou 516200,China
  • Online:2020-12-25 Published:2021-01-19
  • Contact: LING Xukun E⁃mail:lxk6534@163.com

摘要:

目的 探讨miR⁃17⁃5p通过调控肌球蛋白调节轻链相互作用蛋白(myosin regulatory light chain interacting protein,MYLIP)对结直肠癌(colorectal cancer,CRC)细胞增殖和转移的影响。方法 收集CRC 者以及对应的癌旁组织标本和CRC细胞系SW620,Lovo,Caco⁃2,HT29及正常结肠上皮细胞FHC。用脂质体 转染技术将miR⁃17⁃5p inhibitor(实验组)和miR⁃NC(阴性 对照组)瞬时转染CRC细胞,用RT⁃qPCR法检测CRC 组织和细胞中miR⁃17⁃5p的表达水平,双荧光素酶报告基因实验检测MYLIP与miR⁃17⁃5p的靶向关系,采用 MTT法检测CRCLovo细胞的增殖活力,Transwell小室法检测CRCLovo细胞侵袭和迁移能力,Western blotting 法检测 CRC 组织细胞中 MYLIP 及上皮间质转化(epithelial⁃mesenchymal transition,EMT)相关蛋白的表达 水平。结果 CRC组织中miR⁃17⁃5p的表达水平明显高于癌旁组织(P < 0.05),CRCLovo细胞中miR⁃17⁃5p 表达水平最高(P < 0.05)。敲降miR⁃17⁃5p可以抑制CRCLovo细胞的增殖和转移。miR⁃17⁃5p能与MYLIP 3′UTR特异性结合,且miR⁃17⁃5p可以靶向调控MYLIP的蛋白水平。miR⁃17⁃5p靶向下调MYLIP的表达能够 促进CRCLovo细胞的增殖和转移。结论 miR⁃17⁃5p在CRC的发生发展中起重要作用,可通过下调MYLIP 表达促进CRCLovo

关键词:

Abstract:

Objective To investigate the effect of miR⁃17⁃5p on the proliferation and metastasis of colorectal cancer(CRC)cells by regulating MYLIP. Methods The tissues adjacent to the CRCs as well the CRC cell lines SW620,Lovo,Caco⁃2,HT29 and normal colon epithelial cells FHC were collected from the CRC patients. miR⁃17⁃5p inhibitor(as experimental group)and miR ⁃NC(negative control group)were transfected into CRC cells by lipo⁃ some transfection technique. The expression of miR⁃17⁃5p in CRC tissues and cells was detected by RT⁃qPCR,the targeting relationship between MYLIP and miR⁃17⁃5p was detected by double luciferase reporter gene assay. MTT assay was used to detect the proliferation of colorectal cancer Lovo cells,Transwell chamber assay was used to detect the invasion and migration of colorectal cancer Lovo cells and Western blotting was used to detect the expression of MYLIP and EMT(epithelial ⁃mesenchymal transition)related proteins in CRC cells. Results The expression of miR⁃17⁃5p in CRC tissues was significantly higher than that in the adjacent tissues(P < 0.05),and the highest expression of miR⁃17⁃5p was found in Lovo cells of the colorectal cancers(P < 0.05). Knocking down miR⁃17⁃5p inhibited the proliferation and metastasis of Lovo cells in the colorectal cancers. miR⁃17⁃5p was specifically bound to MYLIP′ s 3′ UTR,and mir ⁃ 17 ⁃ 5p targeted the expression of MYLIP′ s protein. Targeted down ⁃ regulation of MYLIP expression by miR⁃17⁃5p promoted the proliferation and metastasis of Lovo cells in the colorectal cancers. Conclusion miR⁃17⁃5p plays an important role in the occurrence and development of colorectal cancer. It can pro⁃ 

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