内源性硫化氢及其合成酶在人成骨肉瘤细胞中表达的研究

Expression of endogenous hydrogen sulfide and its synthases in human osteosarcoma cells

目的观察硫化氢(H2S)及其合成酶胱硫醚-β-合成酶(CBS)、胱硫醚-γ-裂解酶(CSE)和3-巯基丙酮酸硫转移酶(MPST)在人成骨细胞株及人成骨肉瘤细胞系中的表达。方法我科于2013年6月至2014年12月运用免疫组化染色和蛋白质印迹方法在人永久性成骨细胞株h FOB1.19,成骨肉瘤细胞株Saos-2、MG-63和U-2 OS中分别检测CBS、CSE和MPST基因表达量、蛋白含量;通过敏感硫电极法检测H2S在正常成骨组织和骨肉瘤细胞中的含量。分别比较H2S及其合成酶在各组之间的差异,初步探讨H2S及其合成酶与不同成骨肉瘤细胞之间的关系。结果在成骨肉瘤标本和细胞中CBS、CSE和MPST基因的较高表达;成骨肉瘤组织与成骨细胞来源的正常和恶性肿瘤细胞CBS、CSE和MPST基因表达的蛋白水平以及H2S产生率不同,其中成骨肉瘤细胞株U-2 OS表达水平最高。结论内源性H2S及它的合成酶CBS、CSE和MPST存在于人恶性成骨肉瘤组织和细胞中,高水平的H2S及其生物合成物质或许能成为成骨肉瘤治疗的新靶点。

Objective To investigate the expression levels of endogenous hydrogen sulfide（H2S） andits synthases cystathionine β-synthase（CBS）, cystatbionine γ-lyase（CSE） and 3-mercaptopyruvate sulfurtrans-ferase（MPST） in osteosarcoma cells. Methods From June 2013 to December 2014, immunohistochemistryand Western blotting were used to detect CBS, CSE and MPST m RNA and protein expression in h FOB1.19,Saos-2, MG-63 and U-2 OS cells, and H2 S was determined in normal tissue samples and osteosarcoma cells. Results There was a higher expression of CBS, CSE and MPST in osteosarcoma samples and cells. The osteosar-coma tissue and osteoblast-derived normal and malignant cell lines presented different protein levels of CBS,CSE and MPST, among which osteosarcoma cancer cells U-2 OS expressed highest. Conclusion H2 S and itssynthases CBS, CSE, and MPST exist in the human malignant osteoblastic tissues and cells. These findings im-ply that high levels of H2 S and its biosynthesis may be a new target for osteosarcoma treatment.