卡介苗对大鼠膀胱肿瘤细胞及正常移行上皮细胞的早期影响  

Early effects of Bacillus Calmette-Guerin on rat bladder cancer cells and normal transitional epithe-lial cells

作　　者：陈伟[1] 孙立江[2] 张桂铭[2] 赵雷佐[1] 姚如永[3] 赵军[4] 

机构地区：[1]青岛大学医学院,266003 [2]青岛大学医学院附属医院泌尿外科,266003 [3]青岛大学中心实验室,266003 [4]西安交通大学医学院第二附属医院泌尿外科

出　　处：《中华实验外科杂志》2013年第2期326-329,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30672102）;青岛大学医学院附属医院博士科研启动基金资助项目

摘　　要：目的通过卡介苗（BCG）单独刺激膀胱肿瘤细胞和正常膀胱移行上皮细胞及代谢产物作用上述细胞，探讨BCG治疗膀胱肿瘤的机制。方法建立大鼠膀胱肿瘤模型，将大鼠膀胱肿瘤细胞和正常移行上皮细胞原代培养。设普通培养液培养肿瘤细胞（A组），含BCG的培养液培养肿瘤细胞（B组），普通培养液培养正常移行细胞后的上清液培养肿瘤细胞（C组），含BCG培养液培养正常移行细胞后的上清液培养肿瘤细胞（D组）和含BCG培养液培养肿瘤细胞后的上清液培养肿瘤细胞（E组）。酶联免疫吸附剂测定（ELISA）法检测各组细胞上清液中干扰素-y（IFN-1）、肿瘤坏死因子-A（TNF-a）、白细胞介素-10（IL-10）的浓度。流式细胞术（FCM）和末端脱氧核苷酸转移酶介导缺口末端标记法（TUNEL）检测凋亡。结果ELISA检测各组细胞上清液中IFN一叮的浓度无显著改变，波动于（6．5—8．5）ng／L。IL-10在B组[（17．165±3．434）ng／L]，E组[（20．420±2．678）ng／L]水平明显高于其他3组；TNF一0[在B组[（159．887±5．854）ng／L]，D组[（126．227±4．812）ng／L]的浓度较其他3组差异有统计学意义（P〈0．05）。各组细胞均未检测到凋亡的发生。结论BCG作用于肿瘤细胞有抑制生长的作用，但不引发凋亡，肿瘤细胞自身能通过分泌IL-10、TNF—OL参与调节，逃避免疫细胞对其的打击。Objective To explore the mechanism of Bacillus Calmette-Guerin （BCG） for bladder cancer through comparing stimulation of bladder cancer cells and normal bladder transitional epithelial cells with BCG or metabolites. Methods The rat bladder cancer models were established, and the bladder cancer cells and normal transitional epithelial cells were subjected to primary culture. The cancer cells were cultured separately in the normal medium （group A）, the BCG medium （group B）, the supernatant which was obtained from the culture of normal transitional epithelial cells in the normal medium （ group C）, the supernatant which was obtained from the culture of normal transitional epithelial cells in the BCG medium （group D）, and the supernatant which was obtained from the culture of bladder cancer cells in the BCG medium （ group E）. The concentrations of interferon （IFN） -y, tumor necrosis factor （TNF） -a, interleu- kin （IL）-10 in the supernatant of each group were detected by using enzyme-linked immunosorbent assay （ELISA）, and apoptosis was examined by using terminal-deoxynucleo tidyl transferase mediated nick end labeling （TUNEL） and flow cytometry. Results There was no significant difference among groups in the concentrations of IFN-y （6. 5-8.5） ng/L. The concentrations of IL-10 in group B （ 17. 165 ±3. 434） ng/L and group E （20. 420 ±2. 678） ng/L were significantly higher than the rest three groups. The concentra- tions of TNF-a in group B （ 159. 887 ± 5. 854） ng/L and group D （ 126. 227 ± 4. 812 ） ng/L were signifi- cantly different from the other three groups. No apoptosis was detected in any group. Conclusion BCG ean inhibit growth of bladder cancer cells, but cann＇ t induce apoptosis. The bladder cancer ceils can regulate their own growth and escape the attack from the immune cells through the secretion of IL-10, and TNF-a.

关 键 词：卡介苗 膀胱肿瘤 脱噬作用 干扰素-y 肿瘤坏死因子-A 白细胞介素-10 

分 类 号：R737.14[医药卫生—肿瘤]