摘要
目的探讨miR-590-5p通过下游靶基因磷酸酶及张力蛋白同源物基因(phosphatase and tensin homologue deleted on chromosome ten,PTEN)调节人胃癌SGC7901细胞对顺铂敏感性的作用。方法通过Targetscan等生物信息学软件预测miR-590-5p下游靶基因,并用Western blot以及qRT-PCR验证其下游靶基因的表达;通过脂质体Lipofectamine 2000将miR-590-5p mimics及miR-590-5p inhibitor分别转染至人胃腺癌细胞SGC701和SGC7901/DDP,MTT检测转染后的胃癌细胞在不同顺铂浓度(0,2,4,8,16,32μmol/L)时对细胞增殖抑制作用。结果生物信息学软件预测到miR-590-5p与PTEN能够靶向互补。Western blot以及qRT-PCR结果提示,SGC7901细胞和SGC7901/DDP分别过表达和抑制miR-590-5p后PTEN的表达分别相应降低和升高(P<0.05)。人胃腺癌细胞SGC7901上调miR-590-5p mimics后较亲本细胞SGC7901对顺铂的敏感性升高,而转染miR-590-5p inhibitor后的SGC7901/DDP细胞对顺铂敏感性降低(P<0.05)。结论miR-590-5p可能通过调节下游靶基因PTEN的表达而影响胃癌细胞对顺铂的敏感性。
Objective To investigate the effect of miR-590-5p on the sensitivity of human gastric cancer SGC7901 cells to cisplatin by downstream target gene phosphatase and tensin homologue deleted on chromosome ten(PTEN).Methods The target gene of miR-590-5p was predicted by bioinformatics software such as Targetscan,and the expression of downstream target genes was verified by Western blot and qRT-PCR.After miR-590-5p mimics and miR-590-5p inhibitor were transfected into human gastric adenocarcinoma cells SGC701 and SGC7901/DDP by Lipofectamine 2000,respectively,MTT assay was used to detect the cell proliferation of gastric cancer cells at different concentrations of cisplatin(0,2,4,8,16,32μmol/L).Results The bioinformatics software predicted that miR-590-5p and PTEN could be targeted and complementary.Western blot and qRT-PCR suggested that the expression of PTEN was decreased after overexpression of miR-590-5p in SGC7901 cells and elevated after inhibition of miR-590-5p in SGC7901/DDP,respectively(P<0.05).The sensitivity of parental cell SGC7901 to cisplatin was increased after transfected with miR-590-5p mimics,while the sensitivity of SGC7901/DDP cells to cisplatin was decreased after transfected with miR-590-5p inhibitor(P<0.05).Conclusion The miR-590-5p may affect the sensitivity of gastric cancer cells to cisplatin by regulating the expression of downstream target gene PTEN.
作者
陈莹
章海斌
刘玉
丁晓娇
王子安
CHEN Ying;ZHANG Haibin;LIU Yu;DING Xiaojiao;WANG Zian(Department of Oncology,First Affiliated Hospital of Bengbu Medical College,Bengbu 233004,China)
出处
《山西医科大学学报》
CAS
2019年第12期1635-1639,共5页
Journal of Shanxi Medical University
基金
安徽省教育厅自然科学研究重点项目(KJ2017A238)
蚌埠医学院创新计划项目(Byycxz1813)