摘要
目的:构建慢病毒稳定敲减FGF19大肠癌细胞系,探究FGF19敲减对人大肠癌细胞增殖、迁移的影响。方法:通过慢病毒转染大肠癌COLO201和HCT116细胞,分别在适宜病毒转染指数条件下转染并构建稳转株;以嘌呤霉素进行筛选,获得稳定敲减FGF19的细胞株;PCR和Western bloting鉴定COLO201和HCT116稳转细胞株中FGF19的表达情况;Transwell实验检测细胞的迁移活性,CCK-8和克隆形成实验检测细胞的增殖活性。结果:成功构建了稳定敲减FGF19的两株大肠癌细胞。稳定敲减FGF19的HCT116、COLO201细胞(KD组)的FGF19 mRNA水平与空载对照组(NC组)相比明显减少(P=0.0011,P=0.0001);与NC组相比,KD组细胞的FGF19蛋白表达明显下降,差异均具有统计学意义(P=0.001,P=0.0001)。KD组细胞的迁移活性以及增殖能力均相比NC组明显下降(迁移P=0.0017/P=0.0001;CCK-8 P=0.0425/P=0.0334;克隆P=0.0051/P=0.0022)。结论:成功构建FGF19敲减慢病毒载体及大肠癌细胞株,敲减FGF19可使大肠癌细胞的迁移活性明显下降,增殖能力降低,为研究FGF19在大肠癌中的作用奠定了实验基础。
Objective:To investigate the effect of fibroblast growth factor 19(FGF19)knockdown on the proliferation and migration of colorectal cancer cells.Methods:The colorectal cancer cell lines COLO201 and HCT116 were transfected with lentivirus to achieve a stable knockdown of FGF19.The mRNA and protein expression of FGF19 were detected using PCR and Western blotting,respectively.Cell migration activity was assessed using the transwell assay,while cell proliferation was evaluated using the CCK-8 assay and colony formation assay.Results:Two colorectal cancer cell lines with stable knockdown of FGF19 were successfully established.The expression of FGF19 mRNA in the knockdown group was significantly lower than in the control group(P=0.0011,P=0.0001).Similarly,the expression of FGF19 protein was significantly decreased in the knockdown group compared to the control group(P=0.001,P=0.0001).Moreover,the migration activity and proliferation ability of the knockdown group were significantly decreased compared to the control group(Transwell P=0.0017/P=0.0001,CCK-8 P=0.0425/P=0.0334,Colon P=0.0051/P=0.0022).Conclusion:In conclusion,we successfully constructed a lentiviral vector for FGF19 knockdown and established colorectal cancer cell lines with stable knockdown of FGF19.Knockdown of FGF19 led to significantly reduced migration activity and proliferation of colorectal cancer cells.This study provides a basis for further investigation into the role of FGF19 in colorectal cancer.
作者
彭朵
王燕
杨子莎
胡迪
胡新荣
赵毅
PENG Duo;WANG Yan;YANG Zisha;HU Di;HU Xinrong;ZHAO Yi(Basic Medical School,Guangdong Medical University,Guangdong Dongguan 523808,China;School of Medical Technology,Guangdong Medical University,Guangdong Dongguan 523808,China)
出处
《现代肿瘤医学》
CAS
2024年第9期1589-1594,共6页
Journal of Modern Oncology
基金
国家自然科学基金资助项目(编号:81572566)
广东省研究生教育创新计划项目(编号:2022XSLT047,2022SFKC063)。
关键词
成纤维细胞因子19
大肠癌
慢病毒载体
细胞迁移
细胞增殖
fibroblast growth factor 19
colorectal cancer
lentiviral vector
cell migration
cell proliferation
作者简介
彭朵(1996-),女,安徽宿州人,硕士研究生,主要从事分子肿瘤学及临床检验诊断方面的研究工作。E-mail:pengduo0317@gdmu.edu.cn;通信作者:赵毅(1978-),男,黑龙江牡丹江人,博士,副教授,硕士生导师,主要从事分子肿瘤学及代谢重编程方面的研究工作。E-mail:zhaoyicomnet@163.com。
