Xiao Yun, Zhu Li-jin, Jv Li, Qian Ya-ling, Zhang Xing
Institute of Hygiene, Zhejiang Academy of Medical Sciences, Hangzhou 310013, China.
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2011 Oct;29(10):721-5.
To explore the biomarkers and mechanism of kidney toxicity induced by trimethyltin chloride (TMT-Cl) through analyzing the differences of protein expression profiles between vero cells and vero cells exposed to TMT-Cl.
The differences of protein expression levels of three paired samples of vero cells and vero cells exposed to TMT-Cl were compared by two-dimensional gel electrophoresis (2-DE) and liquid chromatography-electrospray ionization-linear trap quadrupole (LC-ESI-LTQ). The differences of expression levels of Annexin A1 and α-Tubulin proteins were validated with western blot assay, and the differences of mRNA expression levels of Annexin A1 and α-Tubulin genes were detected with quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR).
Fifteen spots of differential expression in protein profiles between vero cells and vero cells exposed to TMT-Cl were found, and 9 of these spots were identified by LC-ESI-LTQ. The expression levels of 3 proteins (Annexin A1,similar to RAN protein and a hypothetical protein) increased and the expression levels of 6 proteins(growth factor receptor-bound protein 10, tubulin alpha 6, heterogeneous nuclear ribonucleoprotein, similar to elongation factor SIII p15 subunit, S-adenosylhomocysteine hydrolase and a hypothetical protein) reduced. The expression levels of α-Tubulin protein and mRNA significantly decreased in vero cells exposed to TMT-Cl, as compared with vero cells (P < 0.01). The expression of Annexin A1 protein in all exposure groups was significantly up-regulated, the expression of Annexin A1 mRNA in the groups exposed to 25 and 50 µmol/L TMT-Cl was significantly down-regulated, and The expression of Annexin A1 mRNA in the group exposed to 100 µmol/L TMT-Cl was significantly up-regulated (P < 0.01).
The results of present study suggest that 9 proteins with differential expression detected by LC-ESI-LTQ may be related to the kidney toxicity induced by TMT-Cl, which can serve as the biomarkers of early diagnosis and therapeutic effect for the kidney toxicity induced by TMT-Cl.
通过分析氯化三甲基锡（TMT-Cl）作用前后 vero 细胞蛋白质表达谱的差异，探讨 TMT-Cl 致肾毒性的生物标志物及作用机制。
采用双向凝胶电泳（2-DE）和液相色谱-电喷雾电离-线性阱四极杆质谱联用（LC-ESI-LTQ）技术，比较三对 vero 细胞和 TMT-Cl 染毒 vero 细胞蛋白质表达水平的差异。用蛋白质免疫印迹法验证膜联蛋白 A1（Annexin A1）和α-微管蛋白（α-Tubulin）蛋白表达水平的差异，用定量逆转录聚合酶链反应（qRT-PCR）检测 Annexin A1 和α-Tubulin 基因 mRNA 表达水平的差异。
在 vero 细胞和 TMT-Cl 染毒 vero 细胞的蛋白质图谱中发现 15 个差异表达点，其中 9 个点经 LC-ESI-LTQ 鉴定。3 种蛋白质（Annexin A1、类似 RAN 蛋白和一种假定蛋白）表达水平升高，6 种蛋白质（生长因子受体结合蛋白 10、微管蛋白α6、不均一核核糖核蛋白、类似延伸因子 SIII p15 亚基、S-腺苷同型半胱氨酸水解酶和一种假定蛋白）表达水平降低。与 vero 细胞相比，TMT-Cl 染毒 vero 细胞中α-Tubulin 蛋白和 mRNA 表达水平显著降低（P<0.01）。各染毒组 Annexin A1 蛋白表达均显著上调，25 和 50 μmol/L TMT-Cl 染毒组 Annexin A1 mRNA 表达显著下调，100 μmol/L TMT-Cl 染毒组 Annexin A1 mRNA 表达显著上调（P<0.01）。
本研究结果提示，LC-ESI-LTQ 检测到的 9 种差异表达蛋白质可能与 TMT-Cl 致肾毒性有关，可作为 TMT-Cl 致肾毒性早期诊断及疗效观察的生物标志物。
