CAJ | 학술논문

目的探讨p38丝裂原活化蛋白激酶(MAPK)通路在顺铂诱导正常肝细胞、癌周肝硬化肝细胞和肝癌细胞凋亡过程中的作用.方法应用流式细胞仪、电镜、免疫细胞化学、激光扫描共聚焦显微镜和Western blot检测正常肝细胞株HL-7702、癌周肝硬化肝细胞株QSG-7701和肝癌细胞株QGY-7703加入顺铂和p38MAPK特异性抑制剂SB203580后的凋亡情况,以及p38MAPK、细胞分裂周期25同源体B(CDC25B)、p34cdc2和细胞周期素B1的表达.组间数据的比较采用单因素方差分析和SNK-q检验,P＜0.05为差异有统计学意义.结果正常肝细胞、癌周肝硬化肝细胞和肝癌细胞株加顺铂后,凋亡率均明显增加,以癌周肝硬化肝细胞株最明显(从0.8%增高到41.5%);用顺铂+SB203580处理后,癌周肝硬化肝细胞株的凋亡率降低(28.1%),细胞周期阻滞于S期.电镜、免疫细胞化学、激光扫描共聚焦显微镜均观察到细胞凋亡形态.癌周肝硬化肝细胞分别用顺铂和顺铂+SB203580后,p38MAPK相对表达量分别为0.51±0.05和0.53±0.04,CDC25B相对表达量分别为0.61±0.04和0.59±0.03,p34cdc2相对表达量分别为1.08±0.16和1.21±0.15,与空白对照组(p38MAPK、CDC25B和p34cdc2的相对表达量分别为0.43±0.02、0.28±0.05和1.01±0.12)比较,差异有统计学意义(F=20.056,P＜0.05).结论顺铂通过p38MAPK通路诱导癌周肝硬化肝细胞凋亡,而正常肝细胞和肝癌细胞的凋亡诱导途径可能不同.
목적 탐토p38사렬원활화단백격매(MAPK)통로재순박유도정상간세포、암주간경화간세포화간암세포조망과정중적작용.방법 응용류식세포의、전경、면역세포화학、격광소묘공취초현미경화Western blot검측정상간세포주HL-7702、암주간경화간세포주QSG-7701화간암세포주QGY-7703가입순박화p38MAPK특이성억제제SB203580후적조망정황,이급p38MAPK、세포분렬주기25동원체B(CDC25B)、p34cdc2화세포주기소B1적표체.조간수거적비교채용단인소방차분석화SNK-q검험,P＜0.05위차이유통계학의의.결과 정상간세포、암주간경화간세포화간암세포주가순박후,조망솔균명현증가,이암주간경화간세포주최명현(종0.8%증고도41.5%);용순박+SB203580처리후,암주간경화간세포주적조망솔강저(28.1%),세포주기조체우S기.전경、면역세포화학、격광소묘공취초현미경균관찰도세포조망형태.암주간경화간세포분별용순박화순박+SB203580후,p38MAPK상대표체량분별위0.51±0.05화0.53±0.04,CDC25B상대표체량분별위0.61±0.04화0.59±0.03,p34cdc2상대표체량분별위1.08±0.16화1.21±0.15,여공백대조조(p38MAPK、CDC25B화p34cdc2적상대표체량분별위0.43±0.02、0.28±0.05화1.01±0.12)비교,차이유통계학의의(F=20.056,P＜0.05).결론 순박통과p38MAPK통로유도암주간경화간세포조망,이정상간세포화간암세포적조망유도도경가능불동.
Objective To investigate the roles of p38 MAPK in apoptosis of the normal liver cell, the paratumor cirrhosis hepatocellular cell and the hepatocellular carcinoma cell. Methods Three cell lines were adopted (the normal liver cell line HL-7702, the paratumor cirrhosis hepatocellular cell line QSG-7701 and the hepatocellular carcinoma cell line QGY-7703) and treated with Diamminedichloroplatin (DDP, cisplatin)and p38MAPK inhibitor SB203580. The apoptosis and cell cycles were detected by flow cytometry and electromicroscopy. The expressions of p38MAPK, CDC25B, p34cdc2 and cyclinB 1 were detected by immunocytochemical staining, confocal microscopy and western blot. Results The apoptotic rates in all three cell lines pretreated with DDP increased obviously and the rates in normal liver cells and HCC cells increased continuously even after SB203580 treatment, whereas in paratumor cirrhosis cells the rate decreased and the cell cycle stopped at S phase. Conclusions Cisplatin induces apoptosis in the paratumor cirrhosis hepatocellular cell line QSG-7701 via activation of p38MAPK pathway and it differs in the normal liver cells from the hepatocellular carcinoma cells.