MicroRNA-145 Inhibits Cell Migration and Invasion and Regulates Epithelial-Mesenchymal Transition (EMT) by Targeting Connective Tissue Growth Factor (CTGF) in Esophageal Squamous Cell Carcinoma.
MicroRNA-145 Inhibits Cell Migration and Invasion and Regulates Epithelial-Mesenchymal Transition (EMT) by Targeting Connective Tissue Growth Factor (CTGF) in Esophageal Squamous Cell Carcinoma.
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DOI：
10.12659/msm.897663
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发表时间：
2016-10-23
期刊：
影响因子：
--
通讯作者：
Chen H
中科院分区：
文献类型：
--
作者：
Han Q;Zhang HY;Zhong BL;Wang XJ;Zhang B;Chen H
本研究探讨miR-145靶向结缔组织生长因子（CTGF）对食管鳞癌细胞增殖、迁移、侵袭和上皮间质转化（EMT）的影响机制。共收集了50例ESCC组织及其相应的正常相邻食管组织样本。采用实时荧光定量PCR检测miR-145在食管鳞癌临床标本和细胞系中的表达。免疫组化检测CTGF蛋白表达。采用双荧光素酶报告基因分析法检测miR-145对CTGF 3′UTR荧光素酶活性的影响。分别用miR-145 mimics和CTGF siRNA转染Eca 109细胞，通过MTT法、伤口愈合法和Transwell法检测细胞增殖、迁移和侵袭能力的变化。Western blotting检测EMT相关标志基因的表达。miR-145在食管鳞癌组织和细胞系中的表达明显低于正常组织和细胞系（P<0.05）。食管鳞癌组织中CTGF蛋白的阳性表达率明显高于癌旁正常食管组织（P<0.01）。双荧光素酶报告基因检测结果显示，miR-145可与CTGF的3′UTR特异性结合，对CTGF的荧光素酶活性有55%的抑制作用（P<0.01）。上调miR-145或下调CTGF可抑制食管鳞癌细胞的增殖、迁移、侵袭和EMT过程。MiR-145在ESCC组织和细胞系中明显下调，而CTGF的蛋白表达则表现出相反的趋势。MiR-145通过靶向调节CTGF表达抑制ESCC细胞的增殖、迁移、侵袭和EMT过程。
This study investigated the mechanism of miR-145 in targeting connective tissue growth factor (CTGF), which affects the proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) of ESCC cells. A total of 50 ESCC tissues and their corresponding normal adjacent esophageal tissue samples were collected. Then, miR-145 expression in both ESCC clinical specimens and cell lines was detected using quantitative real-time PCR. CTGF protein was detected using immunohistochemistry. Dual luciferase reporter gene assay was employed to assess the effect of miR-145 on the 3′UTR luciferase activity of CTGF. Eca109 cells were transfected with miR-145 mimics and CTGF siRNA, respectively, and changes in cellular proliferation, migration, and invasion were detected via MTT assay, wound-healing assay, and Transwell assay, respectively. Western blotting assay was used to detect the expression of marker genes related to EMT. MiR-145 was significantly down-regulated in ESCC tissues and cell lines compared with normal tissues and cell lines (P<0.05). We found significantly more positively expressed CTGF protein in ESCC tissues was than in normal adjacent esophageal tissues (P<0.01). Dual luciferase reporter gene assay showed that miR-145 can specifically bind with the 3′UTR of CTGF and significantly inhibit the luciferase activity by 55% (P<0.01). Up-regulation of miR-145 or down-regulation of CTGF can suppress the proliferation, migration, invasion, and EMT process of ESCC cells. MiR-145 was significantly down-regulated in ESCC tissues and cell lines, while the protein expression of CTGF exhibited the opposite trend. MiR-145 inhibited the proliferation, migration, invasiveness, and the EMT process of ESCC cells through targeted regulation of CTGF expression.
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