硫化氢及其合成酶在膀胱癌细胞株中的表达及其作用

doi:10.13481/j.1671-587x.20160501

吉林大学学报(医学版) ›› 2016, Vol. 42 ›› Issue (05): 843-847.doi: 10.13481/j.1671-587x.20160501

• 基础研究 • 下一篇

硫化氢及其合成酶在膀胱癌细胞株中的表达及其作用

宋志强^1,2, 沈海山², 王文佳^1,2, 吴建臣², 李然伟³, 李胜文^1,2

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

收稿日期:2016-03-06 出版日期:2016-09-28 发布日期:2016-09-29
通讯作者: 李胜文,主任医师,教授,博士研究生导师(Tel:010-64308522,E-mail:swli@mails.tsinghua.edu.cn) E-mail:swli@mails.tsinghua.edu.cn
作者简介:宋志强(1984-),男,吉林省汪清县人,在读医学博士,主要从事泌尿系统肿瘤诊断与治疗方面的研究。
基金资助:
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）

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

SONG Zhiqiang^1,2, SHEN Haishan², WANG Wenjia^1,2, WU Jianchen², LI Ranwei³, LI Shengwen^1,2

1. Medical Center, Tsinghua University, Beijing 100084, China;
2. Department of Urology, First Hospital, Tsinghua University, Beijing 100016, China;
3. Department of Urology, Second Hospital, Jilin University, Changchun 130041, China

Received:2016-03-06 Online:2016-09-28 Published:2016-09-29

摘要/Abstract

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 10 μmol·L^-1硫酸氢钠（NaHS）组、50 μmol·L^-1NaHS组、100 μmol·L^-1NaHS组和对照组，MTT法检测24和48 h细胞生存率；②顺铂组（5 μg·L^-1）、顺铂（5 μg·L^-1）+NaHS（100 μmol·L^-1）组，另设无药物处理为对照组，药物处理48 h，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。

关键词: 硫化氢, 胱硫醚&beta, 合成酶, 胱硫醚&gamma, 裂解酶, 膀胱肿瘤

Abstract:

Objective: To explore the expressions of endogenous hydrogen sulfide (H₂S) 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 H₂S 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, 50μmol·L^-1 NaHS group, 100μmol·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 (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 H₂S in the bladder cancer cell lines (5637, T24,UM-UC-3 and EJ) were increased obviously (P<0.05or P<0.01). Compared with control group, exogenous H₂S 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 H₂S and its synthases CBS and CSE have an increased expression level in bladder cancer cell lines compared with the normal bladder cells. H₂S can enhance the proliferation of bladder cancer cells and decrease the apoptosis induced by cisplatin.

Key words: hydrogen sulfide, cystathionine beta synthase, cystathionine gamma lyase, bladder neoplasms

中图分类号:

R737.14

宋志强, 沈海山, 王文佳, 吴建臣, 李然伟, 李胜文. 硫化氢及其合成酶在膀胱癌细胞株中的表达及其作用[J]. 吉林大学学报(医学版), 2016, 42(05): 843-847.

SONG Zhiqiang, SHEN Haishan, WANG Wenjia, WU Jianchen, LI Ranwei, LI Shengwen. Expressions of hydrogen sulfide and its synthases in bladder cancer cell lines and their effects[J]. Journal of Jilin University Medicine Edition, 2016, 42(05): 843-847.

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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

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