|本期目录/Table of Contents|

miRNA-98-5p靶向HMGA2调控结直肠癌细胞增殖、侵袭和上皮间质转化

《现代肿瘤医学》[ISSN:1672-4992/CN:61-1415/R]

期数:
2019年20期
页码:
3565-3571
栏目:
论着(基础研究)
出版日期:
2019-09-08

文章信息/Info

Title:
miRNA-98-5p inhibits proliferation,invasion and EMT of colorectal cancer cells by targeting HMGA2
作者:
李 红;?邓文英;?赵玉州;?臧 凯;?李 璐;?罗素霞
郑州大学附属肿瘤医院(河南省肿瘤医院)内科,河南 郑州 450008
Author(s):
Li Hong;?Deng Wenying;?Zhao Yuzhou;?Zang Kai;?Li Lu;?Luo Suxia
Department of Internal Medicine,the Tumor Hospital Affiliated to Zhengzhou University(Henan Cancer Hospital),Henan Zhengzhou 450008,China.
关键词:
结直肠癌;?微小RNA;?细胞增殖和侵袭;?上皮间质转化;?高迁移率族蛋白2
Keywords:
colorectal cancer;?microRNA;?cell proliferation and invasion;?epithelial-mesenchymal transition (EMT);?high mobility group A2
分类号:
R735.3+5;R735.3+7
DOI:
10.3969/j.issn.1672-4992.2019.20.005
文献标识码:
A
qq自动领红包软件:
目的:研究miRNA-98-5p在结直肠癌细胞中的表达水平及对癌细胞增殖和侵袭的影响,探讨miRNA-98-5p在结直肠癌中的临床意义及可能的分子机制。方法:收集2016年8月至2018年2月手术切除的结直肠癌组织标本60份,实时荧光定量PCR法检测结直肠癌组织和癌旁组织中miRNA-98-5p的表达水平,免疫组化检测分析HMGA2的表达强度。分析miRNA-98-5p与结直肠癌肿瘤生物学特征和HMGA2表达的相关性。实时荧光定量聚合酶链反应(qRT-PCR)检测miRNA-98-5p在结直肠癌细胞中的表达情况;应用miRNA-98-5p模拟物转染人结直肠癌HCT116细胞,CCK-8法检测细胞增殖情况,Transwell小室法检测细胞侵袭情况,Western blot检测HMGA2、E-cadherin和Vimentin蛋白表达。采用双荧光素酶活性实验验证miRNA-98-5p对HMGA2的靶向作用。构建裸鼠皮下移植瘤模型,观察肿瘤生长状况。结果:结直肠癌组织miRNA-98-5p的表达水平低于癌旁组织,结直肠癌组织HMGA2的表达水平高于癌旁组织(P<0.05)。相关性分析显示,HMGA2表达强度与miRNA-98-5p表达水平呈负相关(r=-0.536,P<0.001)。miRNA-98-5p在结直肠癌细胞中表达水平低于对应结直肠黏膜正常细胞(P<0.05);与转染阴性对照细胞比较,转染miRNA-98-5p模拟物的HCT116细胞增殖水平和侵袭能力均受到抑制(P<0.05),HMGA2、Vimentin蛋白表达水平降低,E-cadherin表达增高。双荧光素酶报告基因结果提示HMGA2是miRNA-98-5p的靶基因。裸鼠皮下成瘤实验结果显示与Blank组和NC组相比,实验组肿瘤生长缓慢,重量明显降低。结论:miRNA-98-5p在结直肠癌细胞中表达下调,且miRNA-98-5p通过调节HMGA2的表达从而影响结直肠癌细胞增殖、侵袭和上皮间质转化状态。
Abstract:
Objective:To investigate the role of miRNA-98-5p on the proliferation and invasion of colorectal cancer cells and the possible regulatory mechanisms between miRNA-98-5p and HMGA2.Methods:From August 2016 to February 2018,60 surgical specimens were collected.There expression of miRNA-98-5p and HMGA2 at mRNA level in colorectal cancer tissues and paracancerous tissues were quantified by quantitative real-time PCR.The correlation between miRNA-98-5p and the biological features of colorectal cancer as well as HMGA2 expression was analyzed.The expression of miRNA-98-5p in colorectal cancer was detected by real-time PCR.The miRNA-98-5p was overexpressed by Lipofectamine 3000 transfection with miRNA-98-5p mimics.The effects of miRNA-98-5p on cell proliferation and invasion abilities were detected by CCK-8 assay and Transwell assay.The protein expression of high mobility group A2 (HMGA2),E-cadherin and Vimentin was determined by Western blot.The regulatory mechanism between HMGA2 and miRNA-98-5p in HCT116 cells was detected by dual-luciferase reporter assay.Subcutaneous tumor formation model in nude mice was used to evaluate the effects of miRNA-98-5p on tumor growthin vivo.Results:The expression of miRNA-98-5p was significantly decreased compared with adjacent tissues (P<0.05).The protein expression of HMGA2 was significantly increased compared with adjacent tissues.The HMGA2 and miRNA-98-5p expression were negatively correlated(r=-0.536,P<0.001).The results of CCK-8 assay and Transwell assay showed that overexpression of miRNA-98-5p significantly reduced the proliferation and invasion abilities of colorectal cancer cells (P<0.05).Overexpression of miRNA-98-5p decreased the protein level of HMGA2.Also,it upregulated the expression of E-cadherin and downregulated the expression of Vimentin.The result of dual-luciferase-3' UTR reporter assay confirmed that miRNA-98-5p bound to the 3' UTR of HMGA2.In vivo subcutaneous tumor formation experiment revealed that miRNA-98-5p mimics inhibited the growth and volume of injected tumor.Conclusion:miRNA-98-5p may suppress the colorectal cancer cell proliferation,invasion and epithelial-mesenchymal transition (EMT) by down-regulation of HMGA2.

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备注/Memo

备注/Memo:
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更新日期/Last Update: 1900-01-01