摘要
目的 探讨肺炎衣原体（C.pn）感染在人喉癌HEp-2细胞黏附和迁移中的作用,及其在肿瘤转移中的作用及机制.方法 C.pn增殖培养后感染HEp-2细胞,光镜下观察细胞病变,吖啶橙染色观察细胞内C.pn包涵体的形态特点,透射电镜观察C.pn包涵体超微结构.采用细胞黏附实验观察C.pn感染对HEp-2细胞黏附能力的影响.采用创伤修复实验和Transwell实验检测C.pn感染HEp-2细胞后,细胞迁移能力的改变.结果 C.Pn感染HEp-2细胞72 h后,细胞内的空泡状结构（即包涵体）几乎占据整个胞浆.吖啶橙染色结果显示,细胞内的包涵体呈黄绿色葡萄串状.透射电镜观察显示,包涵体内成熟的梨形原体明显增多,而圆形的网状体则较少.细胞黏附实验结果显示,C.pn感染HEp-2细胞2 h后,C.pn感染组黏附细胞的吸光度（A）值为0.669±0.011,明显高于对照组的A4值（0.558±0.005,P〈0.001）,C.pn感染组的细胞黏附率为119.9%.创伤修复实验结果显示,C.pn感染HEp-2细胞24 h后,细胞向划痕中央迁移的距离明显长于对照组（P〈0.05）.Transwell实验结果显示,C.pn感染HEp-2细胞12 h后,C.pn感染组和对照组的细胞迁移数目分别为（23.40±2.41）个/视野和（10.40±1.67）个/视野,差异有统计学意义（P〈0.001）.结论 C.pn感染能增强HEp-2细胞的黏附能力,同时还能促进HEp-2细胞迁移,提示C.pn感染可能在喉癌转移过程中起促进作用.
Objective To explore the effect of Chlamydia pneumoniae （ C.ph） infection on humam laryngeal carcinoma cell line HEp-2 cell adhesion and migration, to further clarify the role and mechanism of C.pn infection in tumor metastasis.Methods HEp-2 cells were infected with C.pn after the culture and propagation of C.pn.The cytopathic effect was observed by microscopy.Morphological characteristics of C.pn inclusions in HEp-2 cells were examined by fluorescence microscopy and acridine orange staining.The ultrastructural changes of C.pn inclusions in the HEp-2 cells were examined by transmission electron microscopy （TEM）.Cell adhesion assay was performed to investigate the effect of C.pn infection on the adhesion of HEp-2 cells to collagen I.Wound-healing assay and transwell assay were performed to explore the effect of C.pn infection on HEp-2 cell migration.Results At 72 h post-infection, C.pn infected-HEp2 cells were swollen and partially desquamated.Numerous vacuoles （inclusions） were observed and C.pn inclusions occupied almost the whole cytoplasm of the HEp-2 cells.Grape-like C.pn inclusions were observed in the HEp-2 cells stained with acridine orange under a fluorescence microscope at 72 h after infection.Under TEM, there were more mature pear-shaped elementary bodies, but less larger and round reticulate bodies in the HEp-2 cells infected with C.pn for 72 h.In the cell adhesion assay, the A value in C.pn infection group was 0.669 ± 0.011, significantly higher than that in the control group （0.558 ＋0.005） at 2 h after infection （P〈0.001）.The cell adhesion ratio in the C.pn infection group was 119.89%.The migration distance of C.pn infected-HEp-2 cells in the wound-healing assay was significantly longer than that of control cells at 24 h after infection（P〈0.05）.HEp-2 cells infected with C.pn for 12 h migrated more than the control cells in the transwell assay（23.40±2.41 vs 10.40±1.67）（P〈0.001）.Conclusions C.pn infection can significantly promote HEp-2 cell adhesion to collag
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2011年第1期18-22,共5页
Chinese Journal of Oncology
基金
基金项目：国家自然科学基金（NO.30971225）
教育部科学技术研究重点项目（206008）
教育部留学归国人员科研启动基金（2005527）
关键词
喉肿瘤
衣原体
肺炎
细胞黏附
细胞迁移
肿瘤转移
Laryngeal neoplasms Chldmydophila pneumoniae Cell adhesion Cell migration Neoplasm metastasis
