藏花素对膀胱移行细胞癌T24细胞基因表达谱的影响  

Influence of crocin on gene expression profile of human bladder cancer cell lines T24

作　　者：吕纯芳[1] 罗春丽[1] 冀慧莹[1] 赵培[1] 

机构地区：[1]重庆医科大学医学检验系临床检验诊断学省部共建教育部重点实验室,重庆400016

出　　处：《中国中药杂志》2008年第13期1612-1617,共6页China Journal of Chinese Materia Medica

基　　金：重庆市科委自然科学基金(CSTC2005BB5309)

摘　　要：目的:应用基因表达谱芯片研究藏花素(crocin)对膀胱移行细胞癌T24细胞基因表达谱的影响。方法:MTT法测定藏花素对T24细胞的生长抑制率,选出藏花素最佳作用时间和浓度,运用高通量的基因表达谱芯片检测藏花素作用T24细胞前后基因表达谱的变化,筛选差异表达基因。RT-PCR和免疫细胞化学验证上调基因p21^(WAF1)和下调基因cyclinD1。结果:MTT实验结果显示3 mmol·L^(-1)藏花素作用T24细胞48 h的抑制率与对照组相比有显著意义(P<0.05)。基因芯片检测发现3 mmol·L^(-1)藏花素作用T24细胞48 h引起该细胞系基因表达谱广泛地改变,其中表达差异2倍以上的基因共836个,这些差异表达基因的功能涉及多个方面,最为明显的是与细胞生长调节相关的细胞周期调节基因、细胞凋亡调控基因、DNA复制相关基因等类型。RT-PCR检测其中的细胞周期调节相关基因p21^(WAF1)和cyclinD1,结果与基因芯片相符。同时免疫细胞化学结果从蛋白表达水平上佐证p21^(WAF1)和cyclinD1变化与芯片结果一致。结论:藏花素可以诱导T24细胞基因表达谱广泛的改变,并可能通过调控细胞周期相关基因的表达来抑制T24细胞的增殖。Objective： To investigate the changes of gene expression profile in transitional cell carcinoma of bladder T24 cell after crocin treatment, in order to find the possible crocin targets. Method： The bladder cancer T24 cell line was treated with crocin. MTT assay was adopted to determine the inhibition rate for selecting the best effect time and concentration of crocin. Differentially expressed genes on groups with or without treatment of crocin were screened with high throughout cDNA micrearray. One up-regulated gene p21^WAF1 and one down-regulated gone cyclinD1 were selected to undergo analysis by the reverse transcription polymerase chain reaction （RT-PCR）. Moreover, immunocytochemical method was used to evaluate p21^WAF1 and cyclinD1 protein expression. Result： The growth of T24 cells was inhibited remarkably following a marked positive correlation between crocin concentration, time and inhibitor rate. When 3 mmol· L^-1 crocin treated T24 cells for 48h, the difference was significant compared with the control group （P 〈0. 05）. Crocin induced wide changes of the gone expression profile of T24 cells. A total of 836 genes were up-regulated or down-regulated by mere than 2 times, which were involved cell cycle controlling, DNA cell apoptosis, replication factor, and so on. The mRNA expression of p21^WAF1 and cyclinD1 detected by RT-PCR were in accordance with cDNA micrearray data. The results of immunocytochemical method showed that p21^WAF1 and cyclinD1 protein expression were consistent with those mRNA expression. Conclusion： Crocin can induce the significant alteration of gene expression profile of T24 cell. It is suggested that the widly konwn anti-tumor effects of crocin are medicated at least in part by regulating the cell cycle controlling gone expression.

关 键 词：藏花素 膀胱移行细胞癌 基因表达谱芯片 

分 类 号：R285.5[医药卫生—中药学]