具核梭杆菌感染KB细胞影响细胞增殖和炎症因子分泌的研究

摘要/Abstract

摘要： 目的具核梭杆菌（Fusobacterium nucleatum, F. nucleatum），为革兰氏阴性杆菌，是牙周病的重要致病菌之一。研究发现，牙周病原体感染和口腔癌的发生发展密切相关。因此，本研究通过分析细菌感染对KB细胞增殖和细胞因子分泌的影响，为具核梭杆菌感染和口腔癌症发展之间的关系提供新的证据。方法建立具核梭杆菌感染KB细胞模型（MOI=10和100），透射电镜观察细胞和细菌形态变化，PI单染流式细胞术检测细胞周期，ELISA检测上清液中IL6和IL8细胞因子分泌水平。结果透射电镜观察发现，4 h具核梭杆菌侵入细胞，细菌在细胞内保持完整形态，细胞与细胞之间紧密连接破坏。感染24 h后，细胞内仍能发现细菌，菌体被膜状结构包绕，并保持完整形态。低浓度具核梭杆菌（MOI=10）感染KB细胞，细胞周期G1期进程在感染早期加速，细胞增殖率没有显著变化（P> 0.05）。高浓度具核梭杆菌（MOI=100）感染KB细胞，细胞周期S期4 h至8 h细胞数增多，同时促进细胞增殖；感染24 h后抑制G1期进程，S期细胞显著减少，从而抑制细胞增殖（P<0.05）。上清液中IL6和IL8细胞因子分泌水平均升高，并对具核梭杆菌感染的时间和浓度存在依赖性。结论具核梭杆菌感染口腔癌细胞可能通过影响细胞增殖和细胞周期进程，促进细胞炎症反应从而加重癌细胞恶性程度。

关键词: 具核梭杆菌, 细胞周期, 细胞因子, 肿瘤细胞

Abstract: Objective Fusobacterium nucleatum (F. nucleatum), Gramnegative bacteria, is one of the important pathogens of periodontal disease. The studies have found that periodontal pathogen infection and oral cancers development are closely related. Therefore, this study aims to analyze the effects of bacterial infection on cell proliferation and cytokine secretion, providing new evidence for the relationship between F. nucleatum infection and oral cancer development. Methods KB cells infected with F. nucleatum model was established. The morphology of bacteria and cell was observed by transmission electron microscopy. Cell cycle was analyzed by PI staining flow cytometry, the levels of IL6 and IL8 cytokines in the supernatant were detected by ELISA. Results F. nucleatum invaded the cells at 4 h. The bacteria remained intact within the cells, and the tight connection was broken among cells. 24 h after infection, F. nucleatum could still be found within the cells. The bacteria capsule structure wrapped around and maintained integrity. Low concentration of F. nucleatum (MOI=10) infected KB cells promoted G1 phase process at 4 h(P<0.05), but had no significant changes in cell proliferation. High concentration of F. nucleatum (MOI=100) infected KB cells promoted cell proliferation through accelerating S phase cells at 4 h8 h. However, after 24 h infection, the G1 phase was inhibited and the S phase was significantly reduced resulting in cell proliferation blocked (P<0.05). The levels of IL6 and IL8 cytokines in the supernatant increased and the time and concentration were dependent on the KB cells with F. nucleatum. Conclusions Infection of oral cancer cells with F. nucleatum may aggravate the malignant course of cancer cells by affecting cell proliferation and cell cycle progression, exacerbating cell inflammatory responses.

Key words: Fusobacterium nucleatum, cell cycle, cytokines, tumor cell

中图分类号:

R781.4

刘俊超王晴萱张冬梅李琛王宏岩赵海礁潘亚萍. 具核梭杆菌感染KB细胞影响细胞增殖和炎症因子分泌的研究[J]. , 2018, 38(2): 109-113.

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