摘要
目的研究藏红花素(crocin)对人膀胱移行细胞株BIU-87增殖的作用及机制。方法MTT法测定crocin抗增殖效应,FCM分析细胞周期分布,RT-PCR技术测定CyclinD1,p27kip1mRNA表达变化,Westernblot检测CyclinD1,p27kip1蛋白表达变化。结果crocin对细胞生长有抑制作用,且呈时间-剂量依赖关系。FCM检测显示,crocin可使细胞阻滞于G0/G1期;3.2mmol/Lcrocin作用于BIU-87细胞后RT-PCR结果显示,CyclinD1mRNA表达明显下调,p27mRNA无显著变化;Westernblot结果表明CyclinD1蛋白明显下调;p27kip1蛋白表达显著增加。结论crocin对细胞有抗增殖作用,可阻滞细胞于G0/G1期,其可影响CyclinD1mRNA转录,但并不影响p27mRNA表达水平,并调控CyclinD1、p27kip1蛋白表达。
Objeetive To observe the effects of crocin on human bladder cancer BIU-87 cell line and to explore the anti-cancer mechanism. Methods BIU-87 cells of human bladder carcinoma were cultured by cell culture technique. The MTT assay was used to evaluate the inhibitory effect of crocin for the growth of BIU-87 cells. Flow cytometry was used to measure the cell cycle. RT- PCR and Western-blot were used to detect cyclin D1 and p27^kip1 expression. Results The growth of BIU-87 cells was inhibited remarkably in the dosage dependence and time dependence way . Flow cytometry profiles revealed that cells were blocked at G0/G1 phase,the cell proliferation was inhibited obviously. After be treated with 3.2mmol/L crocin, the expression of Cyclin D1 mRNA was down-regulated strikingly,the expression of p27^kip1 mRNA was not with significant difference detected by RT：PCR technique. Cyclin D1 protein expression was down-regulated and p27^kip1 protin was increased detected by Western blot. Conclusion Croein can inhibite the proliferation of BIU-87 ceils effectivelly ,and block ceils at G0/G1 phase . The mechanisms may be changing tumor cell cycles by down-regulating the expression of Cyclin D1 gene and protin,up-regulating the expression of p27^kip1 protin.
出处
《重庆医学》
CAS
CSCD
2008年第2期122-125,共4页
Chongqing medicine
基金
重庆市科委自然科学基金资助项目(CSTC2005BB5309)
