口腔中不同假丝酵母菌的毒力因子活性与口腔鳞状细胞癌相关性研究

Study on virulence factor activity of different Candida strains in oral cavity of patients with oral squamous cell carcinoma and its correlation

目的了解口腔鳞状细胞癌(OSCC)患者口腔中假丝酵母菌的种属分布及其毒力因子活性,以健康人作为对照,分析不同假丝酵母菌的毒力因子活性与OSCC相关性。方法收集100例组织病理学诊断为原发OSCC患者病灶区拭子标本,所有患者无手术、放化疗及其他辅助治疗史。同时收集100例健康者颊黏膜拭子标本为对照组。对所有标本分离培养假丝酵母菌,并采用飞行时间质谱系统对进行鉴定。采用牛血清清蛋白琼脂平板、卵黄琼脂平板、吐温80琼脂平板和血琼脂平板检测假丝酵母菌的蛋白酶、磷脂酶、酯酶、溶血的活性,同时采用结晶紫染色法检测其生物膜的形成。结果 OSCC组白假丝酵母菌59株(72.0%),热带假丝酵母菌8株(9.7%),近平滑假丝酵母菌7株(8.5%),克柔假丝酵母菌5株(6.1%),杜氏假丝酵母菌3株(3.7%);对照组仅检出白假丝酵母菌[20株(83.3%)]和克柔假丝酵母菌[4株(16.7%)]。将OSCC组中分离出的5种假丝酵母菌毒力因子进行比较,不同菌种间的磷脂酶活性、蛋白酶活性、酯酶活性、溶血活性及生物膜形成能力比较,差异均无统计学意义(P>0.05)。结论该研究有助于探究口腔中假丝酵母菌的分布及其毒力因子活性与细胞癌变的相关性,对研究肿瘤机制和临床治疗均有重要意义。

Objective To investigate the species distribution and virulence factor activity of Candides in oral cavity of patients with oral squamous cell carcinoma(OSCC),and to analyze the correlation between virulence factor activity of Candides with OSCC and the control group.Methods Swab samples were collected from 100 patients with primary oral squamous cell carcinoma who were admitted.All patients had no history of surgery, radiotherapy, chemotherapy or other adjuvant treatments, and had been supported by histomopathological diagnosis.Buccal mucosal swab specimens from 100 healthy subjects were collected as control group.All specimens were isolated and cultured, and the colonies were identified by time-of-flight mass spectrometry.The activity of protease, phospholipase, esterase and hemolysis was determined by bovine serum albumin(BSA) AGAR plate, yolk AGAR plate, twain80 AGAR plate and blood AGAR plate.Meanwhile, the formation of biofilm was determined by crystal violet staining.Results In OSCC group, 59 strains of Candida albicans(72.0%),8 strains of Candida tropicalis(9.7%),7 strains of Candida parapsilosis(8.5%),5 strains of Candida krusei(6.1%),and 3 strains of Candida dubliniensis(3.7%) were detected.In the control group, only Candida albicans [20 strains(83.3%)] and Candida krusei[4 strains(16.7%)] were detected.There were no significant differences in phospholipase activity, protease activity, esterase activity, hemolysis activity and biofilm formation ability among five Candida strains isolated from OSCC group(P>0.05).Conclusion This study is helpful to explore oral distribution and virulence factors of Candida in the pattern of activity and correlation, to study the mechanism of tumor and clinical treatment has the important meaning.