硫化氢及其合成酶在膀胱癌细胞株中的表达及其作用  

Expressions of hydrogen sulfide and its synthases in bladder cancer cell lines and their effects

作　　者：宋志强[1,2] 沈海山 王文佳[1,2] 吴建臣 李然伟[3] 李胜文[1,2] 

机构地区：[1]清华大学医学中心,北京100084 [2]清华大学第一附属医院泌尿外科,北京100016 [3]吉林大学第二医院泌尿外科,吉林长春130041

出　　处：《吉林大学学报（医学版）》2016年第5期843-847,共5页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金青年科学基金资助课题(81201527);清华-周大福医学研究专项基金资助课题(202836019-04);清华-裕元医学科学研究基金资助课题(20240000538)

摘　　要：目的:探讨正常膀胱和膀胱癌细胞株中硫化氢(H2S)及其合成酶胱硫醚β合成酶(CBS)和胱硫醚γ裂解酶(CSE)的表达,阐明其在膀胱癌发生发展中的作用。方法:选取膀胱癌5637、T24、EJ、UM-UC-3细胞株和人膀胱永生化上皮SV-HUC-1细胞株,Western blotting检测CBS和CSE蛋白酶表达水平,敏感硫电极法检测H2S产率;选取EJ细胞株进行药物处理,实验分组为,1 10μmol·L^(-1)硫酸氢钠(NaHS)组、50μmol·L^(-1) NaHS组、100μmol·L^(-1) NaHS组和对照组,MTT法检测24和48h细胞生存率;2顺铂组(5μg·L^(-1))、顺铂(5μg·L^(-1))+NaHS(100μmol·L^(-1))组,另设无药物处理为对照组,药物处理48h,MTT法和流式细胞术分别检测各组细胞增殖活性和凋亡率。结果:与SV-HUC-1细胞株比较,膀胱癌5637、T24、EJ和UM-UC-3细胞中CBS和CSE表达及H2S产率均明显增加(P<0.05或P<0.01);外源性H2S可促进EJ细胞增殖,细胞增殖活性随药物剂量增加而升高(P<0.05),且随着药物作用时间的延长而增加(P<0.05)。与顺铂组比较,顺铂联合NaHS组细胞增殖活性明显升高(P<0.05),细胞凋亡率明显下降(P<0.05)。结论:H2S及其合成酶CBS和CSE在膀胱癌细胞株中有表达且高于膀胱正常上皮细胞,H2S促进膀胱癌细胞增殖并降低顺铂的促细胞凋亡作用。Objective：To explore the expressions of endogenous hydrogen sulfide （H2 S）and its synthases cystathionine beta synthase （CBS）and cystathionine gamma lyase （CSE）in the cell lines of normal bladder and bladder cancer,and to clarify their mechanism in the development of bladder cancer.Methods：The bladder cancer cell lines （5637,T24,UM-UC-3,EJ）and human bladder epithelial cell line SV-HUC-1 were selected.The expressions of CBS and CSE in bladder cancer and normal cell lines were analyzed by Western blotting assay and the productivities of H2 S in cell lines were detected by sensitive sulphur electrode assay.The EJ cells were selected based on the previous experimental results and divided into groups as follows：① 10 μmol· L-1 NaHS group, 50 μmol·L-1 NaHS group,100 μmol·L-1 NaHS group and control group.After drug treatment,the cell survival rate was measured by MTT assay at 24 and 48 h.② 5 μg·L-1 cisplatin group,cisplatin （5 μg·L-1 ）＋ NaHS＆amp;nbsp;（100 μmol·L-1 ）group and control group.After medicine treatment,the cell survival rate was measured by MTT assay and the cell apoptotic rate was detected by flow cytometry at 48 h. Results：Compared with the normal bladder cells （SV-HUC-1）,the expression levels of CBS and CSE and the productivity of H2 S in the bladder cancer cell lines （5637,T24,UM-UC-3 and EJ）were increased obviously （P 〈0.05 or P 〈0.01）.Compared with control group,exogenous H2 S promoted the cell proliferation of EJ cells.The cell survival rates were increased with the increase of drug dose （P 〈0.05）,which showed a dose-dependent effect.The cell survival rates were increased with the prolongation of time （P 〈0.05）,which showed a time-dependent effect.After medicine treatment,compared with cisplatin group,the cell viability in cisplatin＋NaHS group was increased （P 〈0.05）and the apoptotic rate was decreased （P 〈0.05）.Conclusion：Endogenous H2 S and its synthases CBS and CSE have an increased expression level in bla

关 键 词：硫化氢 胱硫醚Β合成酶 胱硫醚γ裂解酶 膀胱肿瘤 

分 类 号：R737.14[医药卫生—肿瘤]