CAJ | 학술논문

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.
목적 탐토폐염의원체(C.pn)감염재인후암HEp-2세포점부화천이중적작용,급기재종류전이중적작용급궤제.방법 C.pn증식배양후감염HEp-2세포,광경하관찰세포병변,아정등염색관찰세포내C.pn포함체적형태특점,투사전경관찰C.pn포함체초미결구.채용세포점부실험관찰C.pn감염대HEp-2세포점부능력적영향.채용창상수복실험화Transwell실험검측C.pn감염HEp-2세포후,세포천이능력적개변.결과 C.Pn감염HEp-2세포72 h후,세포내적공포상결구(즉포함체)궤호점거정개포장.아정등염색결과현시,세포내적포함체정황록색포도천상.투사전경관찰현시,포함체내성숙적리형원체명현증다,이원형적망상체칙교소.세포점부실험결과현시,C.pn감염HEp-2세포2 h후,C.pn감염조점부세포적흡광도(A)치위0.669±0.011,명현고우대조조적A4치(0.558±0.005,P<0.001),C.pn감염조적세포점부솔위119.9%.창상수복실험결과현시,C.pn감염HEp-2세포24 h후,세포향화흔중앙천이적거리명현장우대조조(P<0.05).Transwell실험결과현시,C.pn감염HEp-2세포12 h후,C.pn감염조화대조조적세포천이수목분별위(23.40±2.41)개/시야화(10.40±1.67)개/시야,차이유통계학의의(P<0.001).결론 C.pn감염능증강HEp-2세포적점부능력,동시환능촉진HEp-2세포천이,제시C.pn감염가능재후암전이과정중기촉진작용.
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 collagen Ⅰ and migration of HEp-2 cells,indicating that C.pn infection may play an important role in promoting the metastasis of laryngeal cancer.