丝裂霉素联合塞来昔布对膀胱癌T24细胞增殖的影响研究

目的:研究体外丝裂霉素(MMC)联合塞来昔布对浅表性膀胱癌T24细胞增殖的影响及其可能机制。方法:体外培养浅表性膀胱癌T24细胞,分为MMC[0(对照组)、2、5、10、25、50μmol/L]组及其与塞来昔布(50μmol/L)混合组,每个浓度5个复孔,采用MTT法检测作用24h后细胞的增殖抑制率。采用免疫组化法、实时定量聚合酶链式反应法、酶联免疫吸附测定法检测对照组、MMC(50μmol/L)组和混合[塞来昔布+MMC(50μmol/L)]组作用48h细胞中Bcl-2蛋白、作用24h细胞血管内皮生长因子(VEGF)mRNA的表达及作用96h内VEGF蛋白浓度。结果:MMC能明显抑制细胞增殖(P<0.05),且呈浓度依赖性;塞来昔布能明显增强MMC对细胞的增殖抑制作用(P<0.05);与对照组比较,MMC组和塞来昔布+MMC组细胞中Bcl-2蛋白、VEGF mRNA表达均明显降低(P<0.05),且后2组组间比较有统计学差异(P<0.05);塞来昔布+MMC组细胞中VEGF蛋白浓度随作用时间延长明显降低(P<0.01)。结论:塞来昔布可协同增强MMC抑制T24细胞增殖的作用,其机制可能与降低Bcl-2、VEGFmRNA表达水平和抑制细胞分泌VEGF蛋白作用有关。

Effects of Mitomycin Combined with Celecoxib on the Proliferation of Bladder Cancer Cell Line T24

OBJECTIVE： To investigate the effects of mitomycin （MMC） combined with celecoxib on the proliferation of superficial bladder cancer cell line T24 and the possible mechanism. METHODS： The superficial bladder cancer T24 cell line were cultured in vitro, and divided into MMC groups [0 （control group）, 2, 5, 10, 25 and 50 μmol/L] and MMC combined with celecoxib （50 μmol/L） group with each concentration of 5 bores. MTT method was used to detect the proliferation inhibition rate after 24 h. Immunohistochemistry assay, RT-PCR and ELISA method were adopted to detect the expression of Bcl-2 protein after 48 h of treatment and VEGF mRNA after 24 h of treatment and the protein concentration of VEGF within 96 h of treatment in control group, MMC （50 μmol/L） group and mixture [celecoxib＋MMC （50 μmol/L）] group. RESULTS： MMC could inhibit the proliferation of T24 cell （P〈0.05）, in dose dependent manner; celecoxib could significantly enhance the inhibitory effect of MMC （P〈 0.05）. Compared with control group, the expression of Bcl-2 protein and VEGF mRNA were decreased significantly in MMC group and celecoxib＋MMC group （P〈0.05）; there was statistical significance between the latter 2 groups （P〈0.05）. The concentration of VEGF protein in celecoxib＋MMC group was decreased significantly statistically with time （P〈0.01）. CONCLUSIONS： Celecoxib can collaboratively enhance inhibitory effect of MMC on the proliferation of T24 cell. The mechanism may be related to reducing the expression of Bcl-2 and VEGF mRNA and inhibiting the secretion of VEGF protein.