金黄色葡萄球菌L型对胆囊癌细胞的致病变研究

Cytopathic effect of Staphylococcus aureus L-form on human carcinoma cells of gallbladder

目的研究金黄色葡萄球菌L型对胆囊癌细胞(GBC-SD)的致病变作用。方法金葡菌L型感染GBC-SD细胞后,采用光学显微镜、透射电子显微镜及PCR检测其能否侵入细胞,采用四甲基偶氮唑盐微量酶反应比色(MTT)法、增殖细胞核抗原(PCNA)法检测其能否促进细胞增殖,采用基因芯片检测其引起的细胞基因改变。结果PCR检测以及光镜、电镜拍照显示金葡菌L型能够粘附并且进入GBC-SD细胞,MTT法、PCNA检测表明金葡菌L型能够促进GBC-SD细胞增殖。基因芯片结果显示金葡菌L型作用后的GBC-SD细胞有15个基因表达异常,取表达上调基因含表皮生长因子组件粘蛋白类似激素受体2(EMR2)、表达下调基因细胞外基质蛋白2(ECM2)进行RT-PCR法验证,与基因芯片结果符合。结论金葡菌L型能够进入GBC-SD细胞中并促进其增殖,引起细胞EMR2基因上调、ECM2基因下调,提示金葡菌L型可能与胆囊癌的发生、发展存在一定相关性。

To observe the cytopathic effect of Staphylococcus aureus L form on human carcinoma cells of gallbladder （GBC-SD）, these cells were infected with S. aureus L forms. Light and electron microscopy and PCR assay were used to test whether the L-forms could enter into the infected cells and MTT assay was employed to test whether they could promote the proliferation of GBC-SD and production of the proliferating cell nuclear antigens （PCNA） in vivo. In addition, the alteration of cellular genes after action of L-form was determined by means of gene clip. As demonstrated by PCR assay as well as light and electron microscopy, S. aureus L-forms could enter into GBC-SD ceils efficiently. Also, they could improve proliferation of these cells as revealed by MTT assay and testing for PCNA. The result in gene clip showed that 15 gene expression alterations could be demonstrated in GBC-SD ceils infected with S. aureus L-forms. Meanwhile, the result in up-regulation gene of EMR2 （EGF-like module-containing mucin-like hormone-like 2） and down-regulation gene of ECM2 （extracellular matrix protein-g） were matched with those of the gene clip by RT-PCR. It is evident that S. aueus L forms can enter into GBC-SD cells to promote cellular proliferation in vivo and induce up regulation of ERM2 gene and down-regulation of EMC2 gene, suggesting that they may be related to the development and carcinogenesis of gallbladder cancers.