基础医学与临床 ›› 2021, Vol. 41 ›› Issue (7): 1001-1006.

• 研究论文 • 上一篇    下一篇

LINC00607抑制miR-372影响A549/DDP细胞的增殖和凋亡

杨汶川,韩东,杨帆   

  1. 临沂市第三人民医院
  • 收稿日期:2020-08-11 修回日期:2020-12-11 出版日期:2021-07-05 发布日期:2021-06-17
  • 通讯作者: 杨汶川 E-mail:a2kt2i@163.com
  • 基金资助:
    山东省自然科学基金

LINC00607 affects the proliferation and apoptosis of A549/DDP cells through inhibiting miR-372

  • Received:2020-08-11 Revised:2020-12-11 Online:2021-07-05 Published:2021-06-17

摘要: 目的 探讨LINC00607对肺癌顺铂耐药细胞(A549/DDP)增殖、凋亡的影响和可能分子机制。方法 RT-qPCR检测肺癌组织、癌旁组织、A549、A549/DDP中LINC00607和miR-372的表达水平。将A549/DDP分为pcDNA3.1组、pcDNA3.1-LINC00607组、pcDNA3.1-LINC00607+ miR-NC组、pcDNA3.1-LINC00607+ miR-372组。四甲基偶氮唑蓝(MTT)和集落形成实验检测细胞增殖,流式细胞测量术检测细胞凋亡。Western blot检测Cleaved-caspase3和Ki67蛋白表达。双荧光素酶报告基因和RT-qPCR实验确定LINC00607对miR-372的靶向调控作用。结果: 与癌旁组织比较,肺癌组织中LINC00607表达显著降低,miR-372表达显著升高(P<0.05)。与A549细胞比较,A549/DDP细胞中LINC00607表达显著降低,miR-372表达显著升高(P<0.05)。与pcDNA3.1组比较,pcDNA3.1-LINC00607组A549/DDP细胞存活率、克隆形成数、Ki67蛋白表达显著降低,而凋亡率、Cleaved-caspase3蛋白表达显著升高(P<0.05)。与pcDNA3.1-LINC00607+miR-NC组比较,pcDNA3.1-LINC00607+miR-372组A549/DDP细胞存活率、克隆形成数、Ki67蛋白表达显著升高,而凋亡率、Cleaved-caspase3蛋白表达显著降低(P<0.05)。LINC00607靶向负调控miR-372表达。结论 LINC00607通过下调miR-372能够抑制A549/DDP细胞增殖,诱导细胞凋亡。

关键词: LINC00607, miR-372, A549/DDP细胞株, 耐药敏感性

Abstract: Objective To explore the effect of LINC00607 on proliferation and apoptosis of lung cancer cisplatin-resistant cells (A549/DDP) and its possible molecular mechanism. Methods Real-time fluorescence quantitative PCR (RT-qPCR) detected the expression levels of LINC00607 and miR-372 in lung cancer tissues, adjacent tissues, A549, A549/DDP cells. A549/DDP cells were divided into pcDNA3.1 group, pcDNA3.1-LINC00607 group, pcDNA3.1-LINC00607 + miR-NC group, pcDNA3.1-LINC00607 + miR-372 group. Methyl thiazolyl tetrazolium (MTT) and colony formation experiments tested cell proliferation, flow cytometry measured apoptosis, and Western blot detected the expression levels of Cleared-caspase3 and Ki67. Dual luciferase reporter gene and RT-qPCR experiments confirmed the targeted regulation effect of LINC00607 on miR-372. Results Compared with adjacent tissues, the expression of LINC00607 in lung cancer tissues was significantly reduced, while the expression of miR-372 was significantly increased (P<0.05). Compared with A549 cells, the expression of LINC00607 in A549/DDP cells was significantly reduced, while the expression of miR-372 was significantly increased (P<0.05). Compared with pcDNA3.1 group, A549/DDP cell survival rate, clone formation numbers, Ki67 protein expression in pcDNA3.1-LINC00607 group were significantly decreased, and apoptosis rate and Cleared-caspase3 protein expression were significantly increased (P<0.05). Compared with the pcDNA3.1-LINC00607 + miR-NC group, the A549/DDP cell survival rate, clone formation numbers, Ki67 protein expression in the pcDNA3.1-LINC00607 + miR-372 group were significantly increased, and the apoptosis rate, Cleared-caspase3 protein expression were significantly reduced (P<0.05). LINC00607 targets and negatively regulates miR-372 expression. Conclusions LINC00607 can inhibit the proliferation and induce apoptosis of A549/DDP cells by down-regulating miR-372.

Key words: LINC00607, miR-372, A549/DDP cell line, drug resistance sensitivity