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[1]苗 雨,尹 华,张 旭,等.Let-7f对人结肠癌细胞上皮间转换及侵袭能力的影响[J].宁夏医科大学学报,2018,(12):1365-1369.[doi:10.16050/j.cnki.issn1674-6309.2018.12.001]
 MIAO Yu,YIN Hua,ZHANG Xu,et al.Effects of Let-7f on Epithelial-Mesenchymal Transition, Migration and Invasion in Human Colon Cancer Cells[J].Ningxia Medical University,2018,(12):1365-1369.[doi:10.16050/j.cnki.issn1674-6309.2018.12.001]
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《宁夏医科大学学报》[ISSN:1005-8486/CN:64-1029/R]

卷:
期数:
2018年12期
页码:
1365-1369
栏目:
论 著
出版日期:
2019-12-30

文章信息/Info

Title:
Effects of Let-7f on Epithelial-Mesenchymal Transition, Migration and Invasion in Human Colon Cancer Cells
文章编号:
1674-6309(2018)12-1365-05
作者:
苗 雨1 尹 华1 张 旭1 郭 丹2 张飞雄1 阮继刚1
(1. 宁夏医科大学总医院心脑血管病医院消化内科,银川 750004; 2. 宁夏医科大学,银川 750004)
Author(s):
MIAO Yu1 YIN Hua1 ZHANG Xu1 GUO Dan2 ZHANG Feixiong1 RUAN Jigang1
(1. Department of Digestive Internal Medicine,General Hospital of Ningxia Medical University, Yingchuan 750004; 2. Ningxia Medical University,Yinchuan 750004)
关键词:
Let-7f上皮间质转化结肠癌细胞侵袭转移
Keywords:
Let-7f epithelial-mesenchymal transitioncolon cancer cellmigrationinvasion
分类号:
R735.3+5
DOI:
10.16050/j.cnki.issn1674-6309.2018.12.001
文献标志码:
A
摘要:
目的 探讨Let-7f在结肠癌组织和结肠癌细胞株中的表达,分析上调Let-7f对人结肠癌细胞上皮间质转换(epithelial-mesenchymal Transition,EMT)和迁移侵袭能力的影响。方法 采用qRT-PCR检测Let-7f在结肠癌组织与癌旁组织中的表达,并检测Let-7f在结肠癌细胞系(HT29、loVo、SW480、SW620)和人正常结肠上皮细胞(NCM460)中的表达;采用基因转染技术将Let-7f模拟物转染人结肠癌细胞株HT29,qRT-PCR验证转染效率;Western blot检测转染前、后结肠癌细胞中EMT相关分子标志物E-cadherin、N-cadherin和vimentin的表达变化;采用划痕愈合实验、Transwell小室法分别检测转染后的HT29细胞的迁移和侵袭能力变化。结果 qRT-PCR结果显示,Let-7f在结肠癌组织中的表达水平低于癌旁组织(P<0.01);同时,Let-7f在结肠癌细胞系(HT29、loVo、SW480和SW620)中的表达量均低于人正常结肠上皮细胞系(NCM460)(P均<0.01)。转染Let-7f mimic 48h后,qRT-PCR检测转染效率显示:与阴性对照组(NC组)相比,Let-7f mimic转染组Let-7f表达水平升高(P<0.01)。Western blot结果显示,Let-7f mimic转染组细胞中E-cadherin表达水平高于NC组(P<0.05),而N-cadherin和vimentin表达水平低于NC组(P<0.05)。划痕愈合实验显示:Let-7f mimic转染组细胞划痕愈合率低于NC组(P<0.05)。Transwell小室法显示,Let-7f mimic转染组穿过小室的细胞数少于NC组(P<0.05)。结论 Let-7f可抑制人结肠癌细胞上皮-间质转化、迁移侵袭能力。
Abstract:
Objective To investigate the effects and regulation of Let-7f on the epithelial-mesenchymal transition,invasion and migration of colon cancer cells. Methods qRT-PCR was used to detect the expression of Let-7f in colon cancer and adjacent colon tissues and colon cancer cell lines including HT29,loVo,SW480 and SW620, and normal intestinal epithelial cells NCM460. Let-7f mimic was transfected into human colon cancer cell line HT29 by gene transfection technology, and the transfection efficiency was verified by qRT-PCR. Western blot was used to measure the expression changes of EMT-related molecular markers E-cadherin, N-cadherin and vimentin in colon cancer cells before and after transfection. Scratch healing test and Transwell chamber method were used to detect the migration and invasion of HT29 cells after transfection. Results The results of qRT-PCR showed that the expression level of let-7f in colon cancer tissue was lower than that in adjacent tissues(P<0.01),and the expression level of let-7f in colon cancer cell lines (HT29, loVo,SW48 0 and SW620) was lower than that in human normal colon epithelial cell lines(NCM460)(P<0.01). After 48 hours of let-7f mimic transfection, the efficiency of qRT-PCR detection showed that compared with the negative control group (NC group),the expression level of let-7f in the mimic transfection group increased (P<0.01). Western blot showed that the expression of E-cadherin was higher in mimic transfection group than in NC group (P<0.05),while the expression of N-cadherin and vimentin was lower in mimic transfection group than in NC group(P<0.05). Scratch healing test showed that the rate of cell scratch healing in mimic transfection group was lower than that in NC group(P<0.05). Transwell chamber method showed that the number of cells passing through the chamber in mimic transfection group was less than that in NC group (P<0.05). Conclusion Let-7f can inhibit epithelial-mesenchymal transition, migration and invasion of human colon cancer cells.

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备注/Memo

备注/Memo:
收稿日期:2018-08-14
基金项目:国家自然科学基金(8146037);宁夏自然科学基金(NZ13143;NZ15141);宁夏医科大学校级项目(XQ2012008)
作者简介:苗雨(1983-),女,硕士,E-mail:wsmynn@163.com
通信作者:阮继刚(1963-),男,主任医师,硕士研究生导师。 E-mail:rjg0188@163.com
更新日期/Last Update: 2018-12-30