T24膀胱癌细胞ARID1A基因的表达与细胞增殖活力相关性研究  

Expression of ARID1A in T24-9 cell line and its relationship with the proliferative activity

作　　者：吴金斌[1] 漆正宇[2] 晏耀明[1] 翟庆娜[1] 余振东[1] 

机构地区：[1]北京大学深圳医院检验科,广东深圳518036 [2]北京大学深圳医院男性生殖与遗传重点实验室,广东深圳518036

出　　处：《中国实验诊断学》2012年第5期792-794,共3页Chinese Journal of Laboratory Diagnosis

基　　金：深圳市科技计划项目(编号201103036)

摘　　要：目的探讨AT丰富结合域1A(ARID1A)基因的表达与膀胱癌细胞T24增殖活力的关系,为进一步基因功能研究提供基础。方法利用梯度稀释法分离获得膀胱移行癌细胞系T24的高增殖亚克隆细胞株,体外评价细胞株和亲代细胞系增殖活力差异,检测细胞周期调控基因CCND1和膀胱癌肿瘤干细胞标志因子CD44。检测染色质重建复合物SWI/SNF亚基基因ARID1A在亚克隆和亲代细胞系的转录表达水平差异。结果分离得到65个T24的亚克隆细胞株,其中T24-9亚克隆细胞株不同于亲代的梭形形态,呈细小的圆形形态,增殖能力明显强于亲代,CC-ND1、ARID1A和CD44的mRNA表达量均有增加。结论 T24细胞系中的含有高增殖活力的干细胞细胞亚群,染色质重建复合物基因ARID1A转录活跃,可能增加ARID1A突变的风险。Objective To investigate the expression of ARID1A gene associated with the proliferative activity of human the transitional cell carcinoma of the bladder（TCCB） cell line T24.Methods Cell monoclones with different proliferative potential were isolated from their parental cell line T24 by limiting dilution.Then the subclone-evolution and expansion,were identified in vitro via biology methods.To determine the patterns of clonal evolution and the distinct proliferation kinetics of individual,we employed the real-time reverse transcription polymerase chain reaction（RT-qPCR） analysis for quantitative surveillance of the subclones with expresses including CCND1,CD44 and ARID1A.Quantitative data were analyzed using independent samples t test.Results A subelone with high proliferative capability T24-9 from T24 was isolated.The subclone was spheroidal-like morphology,with smaller than T24 which is spindle-shaped with long pseudopodium.The clonality of T24-9 was stronger than parental cell line T24.Growth assay showed that T24-9 grew faster to a higher cell density than the parental cell line T24 in 7 days.RT-qPCR showed a higher transcriptional level of three genes CCND1,CD44 and ARID1A in T24-9 than T24.Conclusion Bladder cancer cell line T24 Contains subpopulation with high proliferative activity in which chromatin remodeling complex subunit ARID1A gene is more active in transcription.

关 键 词：膀胱移行细胞癌 T24 ARID1A 增殖 亚克隆细胞株 

分 类 号：R737.14[医药卫生—肿瘤]