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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。
Objective To study the pathogenic effect of Staphylococcus aureus L-type on gallbladder carcinoma cells (GBC-SD). Methods Staphylococcus aureus L-infected GBC-SD cells were stained with light microscopy, transmission electron microscopy and PCR to detect whether they could invade cells. MTT assay, proliferating cell nuclear antigen (PCNA) method to detect whether it can promote cell proliferation, using gene chips to detect the cellular gene changes caused by it. Results The results of PCR and light and electron microscopy showed that L-form of Staphylococcus aureus could adhere to GBC-SD cells. MTT assay and PCNA showed that L-form of S. aureus could promote the proliferation of GBC-SD cells. Gene chip results showed that Staphylococcus aureus L-type GBC-SD cells have abnormal expression of 15 genes, take up-regulated genes containing epidermal growth factor component mucin similar hormone receptor 2 (EMR2), down-regulated gene extracellular matrix protein 2 (ECM2) for RT-PCR validation, in line with the results of the gene chip. Conclusion Staphylococcus aureus L-type can enter into GBC-SD cells and promote its proliferation, resulting in upregulation of EMR2 gene and down-regulation of ECM2 gene, suggesting that L-form of S. aureus may have some correlation with the occurrence and development of gallbladder carcinomas.