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

发布时间：2018-04-14 07:40

本文选题：结直肠肿瘤 + KISS基因　；参考：《吉林大学学报(医学版)》2017年03期

【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KISS1 mRNA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KISS1 mRNA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(P0.05),细胞增殖活力明显降低(P0.05),侵袭能力和迁移能力亦明显下降(P0.05);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(P0.05)。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。
[Abstract]:Aim: to investigate the effect of lentivirus-transfected KISS1 gene overexpression on the proliferation, invasion and migration of colorectal cancer HCT116 cells, and to elucidate its mechanism.Methods: Lentivirus vector was constructed from colorectal cancer cell line with low expression of KISS1 gene and transfected with KISS1 gene. It was divided into three groups: control group, empty vector group (transfected empty vector) and overexpression group (transfected with KISS1 vector).Fluorescence microscopy was used to detect the expression of KISS1 mRNA and protein metastin by real-time PCR and Western blotting, and the ability of cell invasion and migration was detected by CCK-8 assay.Results the expression of KISS1 mRNA and protein in HT29 cells was the lowest, so they were selected as research vectors.After infected with lentivirus containing KISS1 gene, HCT-116 cells could stably express the enhanced green fluorescent protein (EGFP) gene moi 80.Compared with the control group and the empty vector group, the expression of KISS1 mRNA and protein in the overexpression group was significantly higher than that in the control group, and the cell proliferation activity was significantly decreased, the invasion ability and migration ability were also significantly decreased, and the proliferative activity of the empty vector group was significantly decreased compared with the control group, and the cell proliferation activity of the empty vector group was significantly lower than that of the control group.There was no significant difference in invasion ability and migration ability (P 0.05).Conclusion: lentivirus transfection of KISS1 gene can over-express KISS1 protein and inhibit the ability of proliferation, invasion and migration of colorectal cancer cells. The mechanism may be related to the expression of KISS1 protein.
【作者单位】：福建医科大学附属第一医院胃肠外科2区;
【基金】：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)
【分类号】：R735.34

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