[1]刘华松,徐兰兰,张军,等.miRNA-32通过下调PTEN表达促进食管癌细胞的增殖[J].第三军医大学学报,2016,38(22):2438-2443.
 Liu Huasong,Xu Lanlan,Zhang Jun,et al.MicroRNA-32 promotes proliferation via downregulation of phosphatase and tensin homologue (PTEN) in esophageal carcinoma cells[J].J Third Mil Med Univ,2016,38(22):2438-2443.
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miRNA-32通过下调PTEN表达促进食管癌细胞的增殖(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
38卷
期数:
2016年第22期
页码:
2438-2443
栏目:
基础医学
出版日期:
2016-11-30

文章信息/Info

Title:
MicroRNA-32 promotes proliferation via downregulation of phosphatase and tensin homologue (PTEN) in esophageal carcinoma cells
作者:
刘华松 徐兰兰张军 郭家龙林称意原野曾敏程栋梁
十堰市太和医院(湖北医药学院附属医院)胸心大血管外科;湖北医药学院护理学院
Author(s):
Liu HuasongXu Lanlan Zhang Jun Guo Jialong Lin Chengyi Yuan Ye Zeng Min Cheng Dongliang

Department of Thoracic and Cardiovascular Surgery, Shiyan Taihe Hospital, the Affiliated Hospital of Hubei University of Medicine;School of Nursing ,Hubei  University  of  Medicine, Shiyan,Hubei,442000

关键词:
食管癌miR-32磷酸酶张力蛋白基因增殖
Keywords:
esophageal carcinoma miRNA phosphatase and tensin homologue (PTEN) proliferation
分类号:
R394.3;R730.23;R735.1
文献标志码:
A
摘要:

目的      通过检测miR-32在食管癌组织和食管癌细胞系中的表达,探讨miR-32对食管癌增殖和凋亡的影响及分子机制。方法      使用实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)检测47例食管癌组织和7种食管癌细胞系中miR-32表达水平;生物信息学预测miR-32下游靶基因磷酸酶-张力蛋白基因(phosphatase and tensin homologue ,PTEN),并通过荧光素酶报告基因法验证其结合状态;通过miR-32 mimics来研究其对PTEN的 mRNA和蛋白表达的影响及其对食管癌细胞系KYSE-410增殖和凋亡的影响;通过PTEN回补实验进一步验证miR-32对其的作用。结果     89%(42/47)的食管癌肿瘤组织中miR-32表达明显上调,在低分化的食管癌细胞系KYSE-410中表达程度最高(P<0.05);生物信息学预测miR-32可靶向作用于PTEN,高表达miR-32可明显抑制PTEN 3’UTR荧光素酶活性(P<0.05);高表达miR-32可明显促进KYSE-410增殖,并抑制其凋亡,抑制率28%(P<0.05);高表达miR-32对PTEN mRNA表达无明显影响(P>0.05),但可明显抑制其蛋白表达,抑制率48%(P<0.05);相较于miR-32单独处理组,PTEN补救实验可部分恢复PTEN蛋白的表达,同时也可逆转其对凋亡的抑制作用,此外,在96 h组可明显抑制食管癌细胞增殖(P<0.05),在其他时间点与miR-32组无明显差异(P>0.05)。结论     miR-32可通过下调PETN的表达促进食管癌细胞的增殖。

Abstract:

Objective      To investigate the effect and mechanism of microRNA-32 (miR-32) regulation on the proliferation and apoptosis of esophageal carcinoma(EC) cell line KYSE-410. Methods      Quantitative real-time PCR (qRT-PCR) was used to detect the expression level of miR-32 in EC tissues of 47 patients and 7 EC cell lines. Dual-luciferase reporter assay was used to exam the binding site of 3′ UTR-PTEN to miR-32, which PTEN was theoretically predicted as downstream target of miR-32 by TargetScan database. KYSE-410 cells were either transfected with miR-32 mimics alone or co-transfected with miR-32 and constructed PTEN plasmid for PTEN rescue experiment. The regulation role of miR-32 on PTEN mRNA and protein expression, proliferation and apoptosis were assessed by Western blotting, MTT assay and Annexin V-FITC apoptosis detection kit, respectively. PTEN rescue experiment was used to validate the effect of miR-32 on PTEN expression.  Results       The expression of miR-32 was significantly increased in 89% (42/47 patients) EC tissues compared with corresponding adjacent normal tissues, and was highest in low differentiated KYSE-410 cells (P<0.05). Dual-luciferase reporter assay showed that upregulation of miR-32 decreased luciferase activity of 3′ UTR-PTEN, suggesting that PTEN was the functional downstream target of miR-32. In KYSE-410 cells, overexpression of miR-32 promoted the proliferation, inhibited the apoptosis (inhibitory rate 28%, P<0.05), and suppressed PTEN protein expression (inhibitory rate 48%, P<0.05), but had no change in mRNA level. Compared to cells transfected with miR-32 mimics alone, co-transfected cells (PTEN+miR-32 mimics) could partly reverse the inhibitory effect of miR-32 on PTEN protein expression and apoptosis. The proliferation rate was significantly decreased at 96 h, and no differences were seen at other measured time points between PTEN+miR-32 and miR-32 transfected KYSE-410 cells. Conclusions       MiR-32 can promote the proliferation of esophageal carcinoma at least in part by suppression of PTEN expression.

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更新日期/Last Update: 2016-11-23