慢病毒转染KISS1基因对人结直肠癌HCT116细胞增殖、侵袭和迁移能力的影响

doi:10.13481/j.1671-587x.20170322

吉林大学学报(医学版) ›› 2017, Vol. 43 ›› Issue (03): 577-581.doi: 10.13481/j.1671-587x.20170322

慢病毒转染KISS1基因对人结直肠癌HCT116细胞增殖、侵袭和迁移能力的影响

陈志华, 林素勇, 韩宏景, 苏小宝, 陈绍勤, 戴起宝

福建医科大学附属第一医院胃肠外科2区, 福建福州 350005

收稿日期:2016-07-12 出版日期:2017-05-28 发布日期:2017-06-01
通讯作者: 戴起宝,主任医师,硕士研究生导师(Tel:0591-87982082,E-mail:daijin@yahoo.com) E-mail:daijin@yahoo.com
作者简介:陈志华(1986-),男,福建省仙游县人,主治医师,医学硕士,主要从事胃肠道肿瘤基础研究和外科营养应用方面的研究。
基金资助:
福建省教育厅中青年教师教育科研项目资助课题（JB13386）；福建医科大学苗圃基金项目资助课题（2014MP020）；福建医科大学学科带头人培养专项基金资助课题（JXK201303）

Effects of KISS1 gene transfected by lentivirus on proliferation, invasion, and migration abilities of human colorectal cancer HCT116 cells

CHEN Zhihua, LIN Suyong, HAN Hongjing, SU Xiaobao, CHEN Shaoqin, DAI Qibao

Department of Gastrointestinal Surgery, First Affiliated Hospital, Fujian Medical University, Fuzhou 350005, China

Received:2016-07-12 Online:2017-05-28 Published:2017-06-01

摘要/Abstract

摘要： 目的：探讨慢病毒转染KISS1基因过表达对结直肠癌HCT116细胞增殖、侵袭和迁移能力的影响，并阐明其作用机制。方法：选取KISS1基因低表达结直肠癌细胞株，构建慢病毒载体并转染KISS1基因，分为对照组（PBS处理）、空载体组（转染空载体）和过表达组（转染含KISS1载体）。荧光显微镜检查转染的荧光率（MOI），Real-time PCR和Western blotting法检测各组细胞中KISS1mRNA和蛋白（metastin）的表达量，采用CCK-8法检测各组细胞增殖活力，Transwell小室法检测各组细胞侵袭和迁移能力。结果：LoVo、SW620 、SW480、HCT-116和HT29细胞中，HCT-116细胞中KISS1mRNA和蛋白表达量相对最低，因此筛选其作为研究载体。包装有KISS1基因的慢病毒感染HCT-116细胞后，能够稳定表达增强绿色荧光蛋白（EGFP）基因，MOI均>80%。与对照组和空载体组比较，过表达组细胞中KISS1 mRNA和蛋白表达量明显升高（P<0.05），细胞增殖活力明显降低（P<0.05），侵袭能力和迁移能力亦明显下降（P<0.05）；与对照组比较，空载体组细胞增殖活力、侵袭能力和迁移能力差异均无统计学意义（P>0.05）。结论：慢病毒转染KISS1基因能够过表达KISS1蛋白及抑制结直肠癌细胞增殖、侵袭及迁移能力，其机制可能与KISS1蛋白表达有关。

关键词: 结直肠肿瘤, 转染, 侵袭, 转移, KISS1基因

Abstract: Objective: To explore the effects of KISS1 gene transfected by lentivirus on the proliferation, invasion and migration abilities of the colorectal cancer HCT116 cells,and to clarify their mechanisms. Methods: The human colorectal cancer cells with the lowest expression level of KISS1 gene were selected. The lentiviral vectors were builted and transfected the KISS1 gene, and the cells were divided into control group (treated with PBS), empty vector group (treated with empty vector) and over-expression group(treated with KISS1 gene vector).The multiplicity of infection (MOI) of the cells was detected by fluorescence microscope.Real-time PCR and Western blotting methods were used to detect the expression levels of KISS1 mRNA and protein(metastin);CCK-8 method was used to detect the proliferation ability of the cells; Transwell chambers method was used to detect the invasion and migration abilities of the cells. Results: Among LoVo, SW620, SW480, HCT-116, and HT29 cells, the expression levels of KISS1 mRNA and protein were lowest in HCT116 cells, so they were chosen as the research carrier. After transfected with lentiviral vectors, the HCT116 cells could stably express the enhanced green fluorescent protein(EGFP) gene, and the MOI was over 80%. Compared with control group and empty vector group, the expression levels of KISS1 mRNA and protein in the cells in over-expression group were significantly increased (P<0.05); the proliferation abilities of the cells in over-expression group were decreased (P<0.05); the invasion ability and migration ability of the cells in over-expression group were decreased (P<0.05). But the differences of proliferation ability,invasion ability and migration ability of the cells between control group and empty vector group were not statistically significant (P>0.05). Conclusion: The KISS1 gene transfected by lentivirus vector can over-express KISS1 protein and inhibit the proliferation, invasion and migration abilities of the colorectal cancer cells, and the mechanism may be related to the expression of KISS1 protein.

Key words: KISS1, colorectal neoplasms, invasion, metastasis, transfection

中图分类号:

R735.37

陈志华, 林素勇, 韩宏景, 苏小宝, 陈绍勤, 戴起宝. 慢病毒转染KISS1基因对人结直肠癌HCT116细胞增殖、侵袭和迁移能力的影响[J]. 吉林大学学报(医学版), 2017, 43(03): 577-581.

CHEN Zhihua, LIN Suyong, HAN Hongjing, SU Xiaobao, CHEN Shaoqin, DAI Qibao. Effects of KISS1 gene transfected by lentivirus on proliferation, invasion, and migration abilities of human colorectal cancer HCT116 cells[J]. Journal of Jilin University Medicine Edition, 2017, 43(03): 577-581.

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慢病毒转染KISS1基因对人结直肠癌HCT116细胞增殖、侵袭和迁移能力的影响

Effects of KISS1 gene transfected by lentivirus on proliferation, invasion, and migration abilities of human colorectal cancer HCT116 cells

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