长链非编码RNA MLK7-AS1在结肠癌中的表达及其对结肠癌细胞增殖的影响
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刘海鹰, Email: alylhy@aliyun.com

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国家自然科学基金资助项目(81872035)。


Expression of long non-coding RNA MLK7-AS1 in colon cancer and its influence on proliferation of colon cancer cells
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    摘要:

    背景与目的:长链非编码RNA(lncRNA)MLK7-AS1是一种新发现的与多种肿瘤发生发展密切相关的lncRNA,但其在结肠癌中的表达及其作用尚未见报道。因此,本研究旨在探讨lncRNA MLK7-AS1在结肠癌中的表达以及其对结肠癌细胞生物学行为的影响。
    方法:用qRT-PCR检测lncRNA MLK7-AS1在21对结肠癌组织与相对应的癌旁组织手术标本以及在不同结肠癌细胞系(SW480、HCT116、SW620、DLD1、HT29、LOVO)与正常结肠上皮细胞(NCM460)中的表达。将结肠癌细胞转染lncRNA MLK7-AS1过表达质粒后,分别用MTS、平板克隆试验、流式细胞术、Western blot及qRT-PCR分别检测细胞活力、克隆形成能力、细胞周期以及细胞周期相关蛋白表达水平的变化。
    结果:lncRNA MLK7-AS1在结肠癌组织中的表达水平明显高于癌旁正常组织;在各结肠癌细胞株中表达水平明显高于正常结肠上皮细胞(均P<0.05)。将lncRNA MLK7-AS1表达量相对较低的SW480与HCT116转染lncRNA MLK7-AS1过表达质粒后,两种细胞的细胞活力和克隆形成能力均显著增强(均P<0.05);G1期细胞比例明显减少,S期细胞比例明显增加(均P<0.05);细胞周期相关蛋白cyclin D1和CDK6的表达水平均明显上升(均P<0.05)。
    结论:lncRNA MLK7-AS1在结肠癌中高表达,其高表达可以促进结肠癌细胞增殖,其机制可能与上调细胞周期相关蛋白cyclin D1及CDK6表达有关。

    Abstract:

    Background and Aims: Long non-coding RNA (lncRNA) MLK7-AS1 is a newly identified lncRNA that is closely related to the occurrence and development of various tumors. However, the expression and function of lncRNA MLK7-AS1 in colon cancer have not been reported yet. Therefore, this study was conducted to investigate the expression of lncRNA MLK7-AS1 in colon cancer and its effects on the biological behaviors of the colon cancer cells. 
    Methods: The lncRNA MLK7-AS1 expressions in the surgical specimens of 21 paired colon cancer tissue and corresponding adjacent normal tissue, as well as in different colon cancer cell lines (SW480, HCT116, SW620, DLD1, HT29 and LOVO) and normal colon epithelial cell line (NCM460) were detected by qRT-PCR. In colon cancer cells after transfection with lncRNA MLK7-AS1 overexpression plasmid, the changes in cell viability, colony forming capacity, cell cycle and the expressions of cell cycle-associated proteins were determined by MTS assay, plate cloning assay, flow cytometry and Western blot assays as well as qRT-PCR, respectively.
    Results: The expression levels of lncRNA MLK7-AS1 in colon cancer tissue was significantly higher than that in normal adjacent tissue, and in all studied colon cancer cell lines were significantly higher than that in the normal colonic epithelial cells (all P<0.05). In both SW480 and HCT116 cells that had relatively low lncRNA MLK7-AS1 expressions after transfection with lncRNA MLK7-AS1 overexpression plasmid, the cell viabilities and colony forming capacities were significantly enhanced (both P<0.05); the proportion of cells in G1 phase were significantly decreased and the proportion of cells in S phase were significantly increased (all P<0.05); the expression levels of cyclin D1 and CDK6 were significantly up-regulated (all P<0.05).
    Conclusion: LncRNA MLK7-AS1 is highly expressed in colon cancer, which can promote the proliferation and colony formation of colon cancer cells, and the mechanism is probably related to its regulating the cell cycle-associated proteins cyclinD1 and CDK6 expressions.

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蓝俊松, 李楠, 黄志良, 沈西凌, 刘海鹰.长链非编码RNA MLK7-AS1在结肠癌中的表达及其对结肠癌细胞增殖的影响[J].中国普通外科杂志,2020,29(4):420-428.
DOI:10.7659/j. issn.1005-6947.2020.04.005

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  • 收稿日期:2019-04-19
  • 最后修改日期:2019-12-06
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  • 在线发布日期: 2020-04-25