| 摘要: |
| 摘要：目的 研究吡柔比星(pirarubicin,THP)对膀胱癌细胞增殖、凋亡的影响及其作用机制。方法 用THP处理细胞后，流式细胞术（Flow cytometer，FCM）检测细胞周期，以噻唑蓝（Methyl thiazolyl tetrazolium，MTT）测细胞抑制率，QRT-PCR，RT-PCR检测磷脂酶Cε（Phospholipase C epsilon，PLCε）mRNA及凋亡调控基因BCL-2 mRNA的表达，Western blot 检测PLCε的蛋白表达量；将THP和PLCε干扰腺病毒载体（Ad-shPLCε）联合处理细胞，分为空白组，THP处理组，Ad-shPLCε处理组，THP联合Ad-shPLCε处理组，MTT检测各处理组细胞抑制率，Western blot 检测各处理组BCL-2的蛋白表达量。结果 在膀胱癌细胞T24，BIU-87中，吡柔比星对细胞的抑制率呈浓度，时间依赖性，即随着药物浓度增加，处理时间延长，细胞抑制率升高，即1.6和3.2mg /LTHP药物浓度的抑制率高于0.4和0.8mg /L THP( P ＜ 0. 05)，并且药物处理细胞72 h的细胞抑制率高于24h和48 h ( P ＜ 0. 05)；吡柔比星处理细胞后，T24，BIU-87细胞凋亡数明显增加（P<0.05）；并且抑制癌基因PLCεmRNA、蛋白，凋亡调控基因BCL-2 mRNA的表达；THP联合Ad-shPLCε处理细胞后, 较THP单独处理组，细胞增殖受到明显抑制，BCL-2蛋白表达量明显下调（P<0.05）。结论 吡柔比星能有效诱导细胞凋亡，抑制膀胱癌细胞的增殖，可能是通过抑制PLCε癌基因及下游BCL-2基因的表达。 |
| 关键词: 吡柔比星 PLCε BCL-2 膀胱癌 细胞凋亡 |
| DOI：10.3724/SP.J.1008.2014.00708 |
| 投稿时间：2013-12-02修订日期：2014-05-14 |
| 基金项目:国家自然科学基金 |
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| Pirarubicin induces apoptosis of bladder cancer cells and inhibits the expression of PLC ε mRNA |
| 杜红飞,ouliping,lvchangkun,songxuedong,fanyanru,罗春丽 |
| (Chongqing Medical University) |
| Abstract: |
| Abstract: Objective to investigate the effects of pirarubicin on proliferation and apoptosis of bladder cancer cells and its mechanism. Methods After treatment with THP, Flow cytometry was used to measure apoptosis of T24 and BIU-87 cells; The effects of pirarubicin on cell proliferation of T24 and BIU-87 cell lines were detected by MTT; QRT-PCR、RT-PCR were used to examine the mRNA expression of PLCε，BCL-2 in T24 and BIU-87 cell lines with or without pirarubicin treatment，the protein expression of PLCε in pirarubicin treatment cells was determined by western blot ; After combination treatment of THP with Ad-shPLCε in T24 and BIU-87 cells, MTT was used to evaluate cells proliferation and the protein expression of BCL-2 was detected by western blot in each group. Results Pirarubicin showed significantly dose- and time-dependent inhibitory effect on the proliferation of T24 and BIU-87 cell lines, and the inhibition ratios of 1.6 and 3.2 mg /L THP were higher than that of 0. 4 and 0.8 mg /L THP ( P ＜ 0. 05),in addition, the inhibition ratios of treatment with THP for 72h were higher than that of 24h and 48h THP( P ＜ 0. 05).Moreover , Cell apoptotic rate increased in cell with pirarubicin treatment（P<0.05）.The mRNA and protein expression of PLCε were suppressed in cell lines after treatment with THP. Meanwhile, the expression of BCL-2 at mRNA was repressed by Pirarubicin treatment. Combined treatment of THP with Ad-shPLCε exhibited synergistic inhibitory effects on cells proliferation and BCL-2 protein expression compared with THP treatment group (P<0.05). Conclusions Pirarubicin induced cell apoptosis and inhibited cell proliferation, which may be through suppress the expression of PLCε and BCL-2 in bladder cancer cell lines. |
| Key words: pirarubicin PLCε BCL-2 bladder cancer apoptosis |
