Effects of miR-195 suppresses proliferation,metastasis and invasion on non-small cell lung cancer cells by targeting CHEK1

LUO Lei; YANG Yan-hui; LI Ji; WANG Yi; LI Xiao-liang; XIE Xiao-yang

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Effects of miR-195 suppresses proliferation,metastasis and invasion on non-small cell lung cancer cells by targeting CHEK1

更新时间：2023-05-22

- Effects of miR-195 suppresses proliferation,metastasis and invasion on non-small cell lung cancer cells by targeting CHEK1
- null Issue 4, Pages: 261(2019)
- 作者机构：
  
  内江市第一人民医院胸外科,四川,内江,641000
- 作者简介：
- 基金信息：
- DOI：
  CLC：
- Published：2019，
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LUO Lei, YANG Yan-hui, LI Ji, et al. Effects of miR-195 suppresses proliferation,metastasis and invasion on non-small cell lung cancer cells by targeting CHEK1. [J]. null (4):261(2019)
DOI：

LUO Lei, YANG Yan-hui, LI Ji, et al. Effects of miR-195 suppresses proliferation,metastasis and invasion on non-small cell lung cancer cells by targeting CHEK1. [J]. null (4):261(2019) DOI：

摘要

目的研究miR-195靶向调控细胞周期检测点激酶1（CHEK1）对非小细胞肺癌细胞增殖、凋亡、侵袭和转移能力的影响。方法选取非小细胞肺癌A549、H1299、H197细胞株为研究对象，分别转染miR-195及RNA阴性对照序列。采用实时荧光定量PCR法检测细胞中miR-195的表达水平；MTT检测转染miR-195对肿瘤细胞分裂增殖能力的影响；Transwell侵袭实验分析miR-195对细胞侵袭能力的作用；细胞划痕实验检测miR-195对细胞迁移能力的作用；流式细胞仪检测并评价转染miR-195对肿瘤细胞分裂周期的影响；Western blot探究miR-195对肿瘤细胞CHEK1表达的调节；采用Spearman相关分析法分析miR-195及CHEK1表达的相关性。结果 MTT实验表明高表达miR-195可明显抑制肺癌A549、H1299、H1975细胞的体外增殖能力；Transwell侵袭实验结果提示转染miR-195可导致肺癌A549、H1299、H1975细胞侵袭能力明显降低；流式细胞术结果表明，转染miR-195后A549、H1299、H1975细胞系的G1和G2期细胞增加，S期细胞减少；划痕实验结果表明，转染miR-195可以抑制细胞的迁移能力；Western blot结果显示，转染miR-195后A549、H1299、H1975细胞中CHEK1蛋白水平显著降低，上述差异均具有统计学意义(P＜0.05)。Spearman相关分析显示细胞miR-195 mRNA水平与CHEK1蛋白含量呈负相关(r=-0.464 8，P=0.00)。结论 miR-195可靶向调控CHEK1抑制非小细胞肺癌细胞增殖、转移及侵袭。

Abstract

Objective To investigate the effects of miR-195 on the proliferation

apoptosis

invasion and metastasis on nonsmall cell lung cancer (NSCLC) by targetedly suppressed CHEK1. Methods Nonsmall cell lung cancer cell lines A549

H1299 and H197 were transfected with miR-195 and RNAnegative control sequences respectively.The expression of miR-195 in NSCLC cells was detected by real-time fluorescence quantitative PCR. The effect of transfected miR-195 on the proliferation of tumor cells was detected by MTT assay. Cell invasion and metastasis were detected by transwell invasion test and cell scratch test. Cell apoptosis was detected by flow cytometry

the protein levels of CHEK1 were determined by Western blot

the correlation between the expressions of miR-195 and CHEK1 were analyzed by Spearman.Results MTT assay showed that overexpression of miR-195 could significantly inhibite the proliferation of lung cancer cells A549

H1299 and H1975 in vitro. Transwell invasion assay showed that miR195 could significantly reduce the invasive ability of lung cancer cells A549

H1299 and H1975. Flow cytometry showed that the G1 and G2 phase cells of A549

H1299 and H1975 cell lines increased and S phase cells decreased after transfection of miR-195. The scratch test showed that miR-195 could inhibit cell migration.Western blot results showed that the level of CHEK1 protein decreased significantly after transfection of miR-195. The above differences were statistically significant (P<0.05).Spearman correlation analysis showed that there was a negative correlation between the mRNA level of miR-195 and the content of CHEK1 protein (r=-0.464 8

P=0.00). Conclusion miR-195 can inhibit the proliferation

metastasis and invasion of NSCLC cells by targeting CHEK1.

关键词

miR-195CHEK1非小细胞肺癌转移侵袭

Keywords

miR-195CHEK1non-small cell lung cancermetastasisinvasion

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