中国癌症杂志 ›› 2020, Vol. 30 ›› Issue (6): 441-448.doi: 10.19401/j.cnki.1007-3639.2020.06.006

• 论著 • 上一篇    下一篇

miR-203a-3p通过靶向调控GATA6抑制食管鳞癌细胞的增殖和侵袭

殷彩桥 1 ,方呈祥 2 ,陈 婷 1 ,黄 慧 1 ,郑 轶 1 ,谭家武 1 ,张继乔 1 ,张 辉 1   

  1. 1. 湖北民族大学附属民大医院消化内科,湖北 恩施 445000 ;
    2. 湖北民族大学附属民大医院肿瘤科,湖北 恩施 445000
  • 出版日期:2020-06-30 发布日期:2020-07-16
  • 通信作者: 殷彩桥 E-mail: 915683258@qq.com
  • 基金资助:
    中国国家留学基金管理委员会2016年地方合作项目(留金法[2016]5113号,201608420206);湖北民族学院博士启动基金项目(鄂民院科技字[2014]8号,MY2014B021)。

miR-203a-3p inhibits cell proliferation and invasion of esophageal squamous cell carcinoma by targeting GATA6

YIN Caiqiao 1 , FANG Chengxiang 2 , CHEN Ting 1 , HUANG Hui 1 , ZHENG Yi 1 , TAN Jiawu 1 , ZHANG Jiqiao 1 , ZHANG Hui 1   

  1. 1. Department of Gastroenterology, Minda Hospital of Hubei Minzu University, Enshi 445000, Hubei Province, China; 2. Department of Oncology, Minda Hospital of Hubei Minzu University, Enshi 445000, Hubei Province, China
  • Online:2020-06-30 Published:2020-07-16
  • Contact: YIN Caiqiao E-mail: 915683258@qq.com

摘要: 背景与目的:生物信息学分析提示GATA6是miR-203a-3p的潜在靶基因,明确miR-203a-3p通过靶向调控GATA6抑制食管鳞癌细胞的增殖和侵袭。方法:采用Lipofectamine TM RNAiMAX对培养的KYSE-70和KYSE-180细胞瞬时转染。采用实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR)检测miR-203a-3p和GATA6的表达水平。采用蛋白质印迹法(Western blot)检测GATA6蛋白的水平。质粒联合转染后检测相对萤光素酶活性。对食管鳞癌患者标本进行恶性肿瘤与异型增生组织miR-203a-3p和GATA6的表达检测。结果:RTFQ-PCR及Western blot检测结果显示,与对照组比较,GATA6基因和蛋白的表达在miR-203a-3p转染组中降低,在miR-203a-3p inhibitor组却升高,差异均有统计学意义(P<0.05)。与对照组比较,miR-203a-3p转染组KYSE-70细胞增殖能力下降,miR-203a-3p inhibitor组中却升高,差异均有统计学意义(P<0.05)。在KYSE-180中虽然差异无统计学意义,但其趋势却与KYSE-70一致。与对照组比较,在KYSE-70和KYSE-180细胞系中,侵袭细胞数值/视野在miR-203a-3p转染组中均明显下降(P<0.01),在miR-203a-3p inhibitor组中却明显升高(P<0.05)。与miR-203a-3p掠夺型+GATA6野生型组和miR-203a-3p野生型+GATA6突变型组比较,相对萤光素酶活性在miR-203a-3p野生型+GATA6野生型组中降低,差异有统计学意义(P<0.05)。与异型增生组织相比较,100%(10/10)食管鳞癌患者miR-203a-3p在恶性肿瘤组织的表达下调,而GATA6表达水平上调。结论:miR-203a-3p通过靶向调控GATA6抑制食管鳞癌细胞的增殖和侵袭能力。

关键词: 食管鳞状细胞癌, miR-203a-3p, GATA6, 增殖, 侵袭

Abstract: Background and purpose: Bioinformatics analysis showed that GATA6 was a potential target gene for miR-203a-3p. This study aimed to determine whether miR-203a-3p could inhibit proliferation and invasion of esophageal squamous cell carcinoma (ESCC) cells by targeting GATA6. Methods: Transient transfection was performed with Lipofectamine TM RNAiMAX in cultured KYSE-70 and KYSE-180 cell lines. The expressions of miR-203a-3p and GATA6 were detected by real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR). GATA6 protein expression was detected by Western blot. The relative luciferase activity was further detected by dual-luciferase reporter assay after co-transfection with plasmid by FuGENE reagent. The expression levels of miR-203a-3p and GATA6 in esophageal malignant and dysplastic tissues were determined by RTFQ-PCR after microdissection of specimens. Results: The RTFQ-PCR and Western blot results showed that, compared with the control group, the expression levels of GATA6 mRNA and protein were significantly decreased in miR-203a-3p mimic transfection group, while significantly increased in the miR-203a-3p inhibitor transfection group (P<0.05). The expression levels of GATA6 and miR-203a-3p were inversely correlated. Compared with the control group, the cell proliferation viability in the KYSE-70 cell line was decreased in the miR-203a-3p mimic transfection group, while increased in the miR-203a-3p transfection group, which was statistically significant (P<0.05). Although it did not reach statistical significance in the KYSE-180 cell line, its trend was consistent with that of the KYSE-70 cell line. Compared with the control group, the relative number of invasive cell per field in the miR-203a-3p mimic transfection group decreased significantly (P<0.01), while increased in the miR-203a-3p inhibitor transfection group (P<0.05), which had significant statistical significance. Compared with miR-203a-3p scrambled + GATA6 wild type group and miR-203a-3p wild type +GATA6 mutant group, relative luciferase enzyme activity in miR-203a-3p wild type+GATA6 in wild type group significantly decreased (P<0.05). Compared with the dysplastic tissues, the miR-203a-3p expression in esophageal malignant tissues decreased in all the ESCC patients, while GATA6 expression level increased. Conclusion: miR-203a-3p could inhibit the proliferation and invasion ability of ESCC by targeting GATA6.

Key words: Esophageal squamous cell carcinoma, miR-203a-3p, GATA6, Proliferation, Invasion