bcl-2过表达对阿霉素诱导的膀胱癌细胞凋亡和活性氧产生的影响  

Effect of bcl-2 overexpression on adriamycin-induced reactive oxygen species generation and apoptosis in BIU87 bladder carcinoma cells

作　　者：刘奔[1] 孔垂泽[1] 高建[2] 时京[1] 李振华[1] 

机构地区：[1]中国医科大学附属第一医院泌尿外科,沈阳110001 [2]中国医科大学实验技术中心三部

出　　处：《中华实验外科杂志》2006年第10期1167-1169,i0001,共4页Chinese Journal of Experimental Surgery

基　　金：辽宁省自然科学基金(20042082)

摘　　要：目的探讨凋亡相关基因bcl-2过表达对阿霉素(ADR)诱导的膀胱癌细胞凋亡的影响。方法采用脂质体转染法将bcl-2基因转入膀胱癌BIU87细胞,G418筛选获得抗性亚克隆细胞株,逆转录．聚合酶链反应(RT-PCR)法检测bcl-2基因的表达。比较BIU87、BIU87/neo和BIU87/bcl-2细胞对ADR的敏感性。用ADR分别作用于上述细胞24 h,流式细胞仪检测细胞凋亡和细胞内活性氧(ROS)产生情况。结果建立分别稳定表达bcl-2基因、新霉素抗性基因(neo)的膀胱癌亚克隆细胞株BIU87/bcl-2、BIU87/neo。RT-PCR BIU87/bcl-2细胞的bcl-2 mRNA水平显著高于BIU87、BIU87/neo(P＜0．01)。噻唑蓝(MTT)检测显示BIU87/bcl-2细胞对ADR的化疗敏感性明显降低(P＜0．01)。ADR作用24 h后,BIU87、BIU87/neo细胞凋亡率分别为(23．75±3．72)%、(21．94±1．27)%,而BIU87/bcl-2细胞为(3．38±0．95)%(P＜0．01)；ROS分别为(90．45±1．96)%、(88．41±3．88)%和(77．00±3．86)%,BIU87/bcl-2细胞较前两者明显降低(P＜0．01)。结论bcl-2基因能够抑制ADR诱导的膀胱癌BIU87细胞凋亡,降低细胞内ROS水平是其发挥抗凋亡作用的重要机制。Objective To explore the effect of apoptosis-associate gene of bcl-2 overexpression on adriamycin-induced apoptosis in bladder carcinoma cells. Methods Under the induction of liposome, the whole length of bcl-2 cDNA was transfected into human bladder carcinoma cell line BIU87. The subclone cell lines were obtained by persistent G418 selection, bcl-2 gene expression of cancer cells was detected by RT-PCR methods. The growth inhibition effects of adriamycin on cancer cells were observed by tetrazolium bromide colorimetry. Apoptosis rates and intracellular reactive oxygen species （ROS） levels were measured by flow cytometry after cancer cells were treated with adriamycin for 24 h. Results The subclone bladder carcinoma cell lines, stably expressing bcl-2 and neo gene respectively, were successfully selected, named as BIU87/bcl-2 and BIU87/neo. RT-PCR results demonstrated bcl-2 mRNA levels of BIU87/bcl-2 cells were significantly higher than those of BIU87 and BIU87/neo （P〈0.01）. The sensitivity of BIU87/bcl-2 cells to adriamycin was lower significantly than the other two cell lines （P 〈 0.01 ）. After being treated with adriamycin for 24 h, the percentages of apoptotic cells in BIU87 and BIU87/neo were （23.75 ± 3.72） %, （21.94 ± 1.27） % respectively, but those in the B IU87/bcl-2 cells were （3.38 ± 0.95）% （P〈 0.01）. The intracellular ROS levels of BIU87, BIU87/neo and BIU87/bcl-2 were （90.45 ± 1.96） %, （88.41 ± 3.88） % and （77.00 ± 3.86） % respectively, and those of BIU87/bcl-2 cells were lower significantly than those of BIU87 ,BIU87/neo （P〈0.01 ）. Conclusion bcl-2 can block apoptosis induced by adriamycin in BIU87 bladder carcinoma cells and down-regulation of ROS level maybe contribute to this procedure.

关 键 词：膀胱肿瘤 基因 BCL-2 脱噬作用 

分 类 号：R737.14[医药卫生—肿瘤]