CAJ | 학술논문

目的探讨甘露糖敏感性绿脓杆菌制剂(pseudomonasaeruginosa mannose sensitive haemagglutination vaccine,PA-MSHA)对肝细胞癌(hepatocellular carcinoma,HCC)细胞周期的作用及其机制.方法培养人肝癌细胞株MHCC97L及HepG2,以不同剂量的PA-MSHA作用于肝癌细胞.MTT实验检测PA-MSHA对细胞增殖的影响.流式细胞技术检测PA-MSHA对细胞周期的作用.Western blot检测PA-MSHA作用前后细胞周期蛋白Cyclin D1、细胞周期蛋白依赖性激酶2(CDK2)、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)以及细胞周期蛋白激酶抑制剂p21和p27的表达情况.结果与对照组相比,PA-MSHA显著抑制MHCC97L和HepG2细胞的增殖,其作用呈剂量及时间依赖性(P＜0.05).PA-MSHA显著诱导肝癌细胞周期阻滞,PA-MSHA作用后G0-G1期和G2-M期细胞比例显著增高,而S期细胞比例则显著下降(P＜0.05).PA-MSHA显著抑制Cyclin D1、CDK2、PCNA蛋白的表达,而促进p21和p27的表达.外源性甘露糖可显著抑制PA-MSHA对肝癌细胞增殖和周期的作用(P＜0.05).结论 PA-MSHA通过调控细胞周期相关蛋白来诱导HCC细胞周期阻滞,抑制细胞增殖.这一作用是通过肝癌细胞表面的甘露糖的介导实现的.
목적 탐토감로당민감성록농간균제제(pseudomonasaeruginosa mannose sensitive haemagglutination vaccine,PA-MSHA)대간세포암(hepatocellular carcinoma,HCC)세포주기적작용급기궤제.방법 배양인간암세포주MHCC97L급HepG2,이불동제량적PA-MSHA작용우간암세포.MTT실험검측PA-MSHA대세포증식적영향.류식세포기술검측PA-MSHA대세포주기적작용.Western blot검측PA-MSHA작용전후세포주기단백Cyclin D1、세포주기단백의뢰성격매2(CDK2)、증식세포핵항원(proliferating cell nuclear antigen,PCNA)이급세포주기단백격매억제제p21화p27적표체정황.결과 여대조조상비,PA-MSHA현저억제MHCC97L화HepG2세포적증식,기작용정제량급시간의뢰성(P＜0.05).PA-MSHA현저유도간암세포주기조체,PA-MSHA작용후G0-G1기화G2-M기세포비례현저증고,이S기세포비례칙현저하강(P＜0.05).PA-MSHA현저억제Cyclin D1、CDK2、PCNA단백적표체,이촉진p21화p27적표체.외원성감로당가현저억제PA-MSHA대간암세포증식화주기적작용(P＜0.05).결론 PA-MSHA통과조공세포주기상관단백래유도HCC세포주기조체,억제세포증식.저일작용시통과간암세포표면적감로당적개도실현적.
Objective To investigate the effects and mechanism of the pseudomonas aeruginosa mannose sensitive haemagglutination vaccine (PA-MSHA) on the cell cycle of hepatocellular carcinoma (HCC).Methods MHCC97L and HepG2 cells were cultured and treated with different doses of PAMSHA and mannose.MTT and flow cytometry were used to measure cell proliferation and cell cycles,respectively.The expressions of cell cycle related proteins Cyclin D1,CDK2,PCNA,p21,and p27 were evaluated by Western blot.Results Compared with the control group,the proliferation of HCC Cells in the PA-MSHA treated group was significantly inhibited in a time and dose dependant manner (P＜0.05).PA-MSHA significantly arrested HCC cells in the G0-G1 and G2-M phase of the cell cycle,thereby decreasing the proportion of cells in the S phase (P＜0.05).Western blot analysis revealed that PA-MSHA significantly inhibited the expressions of Cyclin D1,CDK2,PCNA,and significantly promoted the expression of p21 and p27.Preincubation of PA-MSHA with mannose significantly inhibited the effects of PA-MSHA on proliferation and cell cycle (P＜0.05).Conclusion By regulating the expression of cell cycle related proteins,PA-MSHA significantly induced HCC cell cycle arrest and inhibited cell proliferation.These effects were mediated through PA-MSHA recognizing and combining to the mannose of HCC cells.