医学分子生物学杂志 ›› 2024, Vol. 21 ›› Issue (1): 25-31.doi: 10.3870/j.issn.1672-8009.2024.01.004

• 论著 • 上一篇    下一篇

miR-200c 靶向 ZEB1 调控宫颈癌细胞的增殖和凋亡 #br#

  

  1. 武汉市红十字会医院急诊科 武汉市, 430015
  • 出版日期:2024-01-31 发布日期:2024-03-07
  • 基金资助:
    湖北省自然科学基金 (No. 2022CFB088)

miR-200c Regulates Proliferation and Apoptosis of Cervical Cancer Cells via Targeting ZEB1 Expression #br#

  1. Emergency Department, Wuhan Red Cross Hospital, Wuhan, 430015, China
  • Online:2024-01-31 Published:2024-03-07

摘要: 目的 探究 miR-200c 对宫颈癌细胞增殖和凋亡的影响及其分子机制方法 利用 RT-qPCR 检测52 例宫颈癌组织及癌旁组织中 miR-200c 的表达水平, 使用 miR-200c 模拟物或抑制物转染宫颈癌细胞系,RT-qPCR 验证转染效率后, 使用 CCK8 检测转染细胞的增殖水平, 流式细胞术检测转染细胞的凋亡水平;生物信息学预测 miR-200c 的靶基因, 荧光素酶报告基因验证其靶向关系; RT-qPCR 检测癌组织中靶基因的表达水平, Pearson 相关性分析 miR-200c 和靶基因的表达关系; RT-qPCR 和蛋白质印迹检测转染靶基因siRNA 或质粒细胞的增殖和凋亡水平结果 宫颈癌组织中 miR-200c 的表达显著低于癌旁组织, 转染 miR-200c 模拟物的 HT3 细胞增殖水平显著降低, 凋亡水平显著增加, 而转染 miR-200c 抑制物的 HeLa 细胞增殖水平显著增加, 凋亡水平显著降低; 荧光素酶报告基因实验证实 ZEB1 miR-200c 的指标靶标, miR-200c模拟物显著降低 ZEB1 表达, 而抑制物则发挥相反效应; 宫颈癌组织中 ZEB1 的表达显著高于癌旁组织, 且其表达与 miR-200c 呈显著负相关关系; 敲低 ZEB1 显著抑制 HT3 的增殖水平, 促进其凋亡, 而过表达ZEB1 则发挥相反效应结论 miR-200c 通过靶向抑制 ZEB1 表达调控宫颈癌细胞的增殖与凋亡, 从而参与宫颈癌的发生发展

关键词: 宫颈癌, miR-200c, ZEB1,  , 增殖,  , 凋亡

Abstract: Objective To study the effect of miR-200c on the proliferation and apoptosis ofcervical cancer cells and its possible mechanism. Methods RT-PCR was used to detect the expression level of miR-200c in 52 cases of cervical cancer tissues and adjacent tissues. The miR-200c mimic or inhibitor was used to transfect cervical cancer cell lines, and the transfection efficiency was verified by RT-PCR. CCK8 assay was used to detect the proliferation activity of transfected cells, while the apoptosis was detected by flow cytometry. The target gene of miR-200c was predicted by bioinformatics analysis, and the dual-luciferase gene reporter assay was applied to verify the targeting relationship. RT-PCR was used to detect the expression level of the target gene in cancer tissuesand adjacent tissues. Pearson correlation analysis between miR-200c and target gene expression wasanalyzed. RT-PCR and Western blotting were used to detect the expression level of target gene incells transfected with siRNA or plasmid. Results The expression level of miR-200c in cervicalcancer tissues was significantly lower than that in adjacent tissues. The proliferation activity of HT3 cells transfected with miR-200c mimics was significantly reduced, and the apoptosis was significantly increased, while the proliferation activity of HeLa cells transfected with miR-200c inhibitors was significantly increased, and the apoptosis of the above cells was significantly reduced. Dual-luciferase gene reporter assay confirmed that ZEB1 is an indicator target of miR-200c, and miR-200c mimics significantly reduced the expression level of ZEB1, while the inhibitor exerted the opposite effect. The expression level of ZEB1 in cervical cancer tissues was significantly higher than that in adjacent tissues, and its expression was significantly negatively correlated with the expression of miR-200c. Knockdown of ZEB1 significantly inhibited the proliferation of HT3 cells and promotedthe apoptosis of cells, while the overexpression of ZEB1 had the opposite effect. Conclusion miR-200c regulates the proliferation and apoptosis of cervical cancer cells by targeting the expression of ZEB1, thereby participating in the occurrence and development of cervical cancer.

Key words: cervical cancer, miR-200c, ZEB1, proliferation, apoptosis

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