摘要
目的探讨敲减KRT19基因表达对膀胱癌T24细胞增殖、凋亡、集落形成的影响。方法对数生长期T24细胞分为shKRT19组和对照组,分别转染shKRT19慢病毒质粒和空载慢病毒质粒,转染48 h,采用实时荧光定量PCR法检测2组细胞KRT19 mRNA相对表达量,采用流式细胞术检测细胞周期、细胞凋亡率;转染后培养0、24、48、72、96 h时采用CCK-8法检测2组细胞增殖率;转染72 h采用克隆形成实验观察2组培养11 d细胞克隆形成数目。结果转染48 h,shKRT19组细胞KRT19 mRNA相对表达量(0.02±0.00)低于对照组(1.00±0.01)(t=169.741,P<0.001),细胞凋亡率[(18.97±2.12)%]高于对照组[(4.89±0.17)%](t=—10.847,P<0.001),G_(0)/G_(1)期[(57.72±3.59)%]、S期[(9.69±0.49)%]、G_(2)/M期[(32.59±3.65)%]细胞比率与对照组[(52.84±1.76)%、(11.01±0.85)%、(36.15±2.29)%]比较差异均无统计学意义(P>0.05)。shKRT19组转染后培养24、48、72、96 h时细胞增殖率[(181.5±2.1)%、(248.9±4.5)%、(337.9±23.4)%、(421.7±3.2)%]均低于对照组[(206.3±4.9)%、(375.4±6.3)%、(546.3±27.9)%、(778.8±4.4)%](P<0.05)。培养11 d,shKRT19组细胞克隆形成数目[(115±3)个]少于对照组[(147±11)个](t=4.861,P<0.001)。结论敲减KRT19基因表达可抑制膀胱癌T24细胞生长、增殖和集落形成,促进细胞凋亡。
Objective To investigate the influences of knockdown of KRT19 gene on the proliferation,apoptosis and colony formation of T24 cells.Methods The T24 cells in logarithmic growth phase were divided into shKRT19 group(transfected with shKRT19 lentivirus plasmid)and control group(transfected with empty lentiviral vector).After 48-h transfection,the relative expression of KRT19 mRNA was detected by real-time fluorescence quantitative PCR.The cell apoptosis rate and cycle were assessed by flow cytometry.The cell proliferation rate was assessed after 24-,48-,72-and 96-h culture by CCK-8 assay in two groups.After 72-h transfection,colony forming was done to observe the number of colony forming cells after 11-d culture.Results After 48-h transfection,the relative expression of KRT19 mRNA was lower in shKRT19 group(0.02±0.00)than that in control group(1.00±0.01)(t=169.741,P<0.001),the apoptosis rate was higher in shKRT19 group[(18.97±2.12)%]than that in control group[(4.89±0.17)%](t=-10.847,P<0.001),and there were no significant differences in the percentages of cells in G_(0)/G_(1),S and G_(2)/M stages between shKRT19 group[(57.72±3.59)%,(9.69±0.49)%,(32.59±3.65)%]and control group[(52.84±1.76)%,(11.01±0.85)%,(36.15±2.29)%](P>0.05).The proliferation rates after 24-,48-,72-and 96-h culture were lower in shKRT19 group[(181.5±2.1)%,(248.9±4.5)%,(337.9±23.4)%,(421.7±3.2)%]than those in control group[(206.3±4.9)%,(375.4±6.3)%,(546.3±27.9)%,(778.8±4.4)%](P<0.05).After 11-d culture,the number of colony forming cells was smaller in shKRT19 group(115±3)than that in control group(147±11)(t=4.861,P<0.001).Conclusion To knockdown KRT19 gene expression in T24 cells can inhibit the cell growth,proliferation and colony formation and promote cell apoptosis.
作者
孔朝辉
王蓬勃
张灏
韩皓名
张汉青
KONG Zhao-hui;WANG Peng-bo;ZHANG Hao;HAN Hao-ming;ZHANG Han-qing(Department of Urology,Henan Provincial People's Hospital,Zhengzhou University People's Hospital,Zhengzhou,Henan 450003,China;Department of Urology,Henan University People's Hospital,Henan Provincial People's Hospital Zhengzhou,Henan 450003,China)
出处
《中华实用诊断与治疗杂志》
2022年第8期761-764,共4页
Journal of Chinese Practical Diagnosis and Therapy
基金
河南省科技攻关计划项目(201004041)。
