LINC00665靶向调控miR-34a-5p对食管癌细胞增殖、迁移和侵袭能力的影响

陈昭伟; 康丽丽; 宋健

doi:10.13898/j.cnki.issn.1000-2200.2024.05.002

LINC00665靶向调控miR-34a-5p对食管癌细胞增殖、迁移和侵袭能力的影响

The effects of LINC00665 targeting miR-34a-5p on the proliferation, migration and invasion of esophageal cancer cells

摘要

摘要:
目的探讨LINC00665靶向调控微小RNA(miR)-34a-5p对食管癌细胞增殖、迁移和侵袭能力的影响。
方法选取食管癌病人肿瘤组织及癌旁组织，采用qRT-PCR法检测组织中LINC00665、miR-34a-5p表达水平。以食管癌细胞株EC9706为研究对象，设置对照组、阴性转染(siNC)组、LINC00665小干扰RNA载体(siLINC00665)组、siLINC00665+inhibitor阴性转染(anti-NC)组、siLINC00665+miR-34a-5p inhibitor(anti-miR-34a-5p)组，CCK-8法检测各组细胞增殖能力；Transwell实验检测细胞侵袭、迁移数；Western blotting检测基质金属蛋白酶2(MMP-2)、细胞周期素D1(CyclinD1)、p21蛋白表达水平；双荧光素酶实验验证LINC00665和miR-34a-5p的靶向关系。
结果食管癌病人肿瘤组织LINC00665表达水平较癌旁组织明显增加，miR-34a-5p表达水平明显下降(P＜0.01)。与对照组、siNC组相比，siLINC00665组细胞OD值(24、48 h)、侵袭和迁移数、CyclinD1、MMP-2蛋白、LINC00665表达均降低，p21蛋白、miR-34a-5p表达均增加(P＜0.05)；与siLINC00665+anti-NC组相比，siLINC00665+anti-miR-34a-5p组细胞OD值(24、48 h)、侵袭和迁移数、CyclinD1、MMP-2蛋白、LINC00665表达均增加，p21蛋白、miR-34a-5p表达均降低(P＜0.05)。双荧光素酶实验证实LINC00665和miR-34a-5p靶向关系。
结论沉默LINC00665可靶向上调miR-34a-5p表达，进而抑制食管癌细胞增殖、迁移和侵袭能力。

Abstract:
Objective To investigate the effects of LINC00665 targeting and regulating micrornas(miR) 34a-5p on the proliferation, migration and invasion of esophageal cancer cells.
Methods The cancer tissues and adjacent tissues of patients with esophageal cancer were collected, the expression levels of LINC00665 and miR-34a-5p in tissues were detected by qRT-PCR.The esophageal cancer cell line EC9706 were divided into the control group, negative transfection(siNC) group, LINC00665 small interfering RNA vector(siLINC00665) group, siLINC00665+inhibitor negative transfection(anti-NC) group and siLINC00665+miR-34a-5p inhibitor(anti-miR-34a-5p) group.The proliferation ability of cells was detected by CCK-8 method, the numbers of cell invasion and migration were detected by Transwell assay; the protein expression levels of matrix metalloproteinase 2(MMP-2), cyclin D1(CyclinD1) and p21 were measured by Western blotting, and the targeting relationship between LINC00665 and miR-34a-5p was verified by dual-luciferase experiment.
Results The expression level of LINC00665 in cancer tissues of patients with esophageal cancer was obviously higher than that in adjacent tissues, and the expression level of miR-34a-5p in cancer tissues was obviously lower (P < 0.01).Compared with the control group and siNC group, the OD value(24, 48 h), numbers of invasion and migration, expression levels of CyclinD1, MMP-2 proteins and LINC00665 in siLINC00665 group were obviously lower, and the expression levels of p21 protein and miR-34a-5p was obviously higher (P < 0.05).Compared with the siLINC00665+anti-NC group, the OD value (24, 48 h), numbers of invasion and migration, expression levels of CyclinD1, MMP-2 proteins and LINC00665 in siLINC00665+anti-miR-34a-5p group were obviously higher, and the expression levels of p21 protein and miR-34a-5p were obviously lower (P < 0.05).The targeting relationship between LINC00665 and miR-34a-5p was confirmed by dual-luciferase reporter gene experiment.
Conclusions Silencing LINC00665 can target and up-regulate the expression of miR-34a-5p, thereby inhibiting the proliferation, migration and invasion of esophageal cancer cells.

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