CAJ | 학술논문

目的研究人肝癌发生过程中p38MAPK和乙型肝炎病毒X蛋白(HBX)对细胞增殖和凋亡的影响,探讨肝癌发生的机制. 方法应用免疫组织化学和DNA原位末端标记(TUNEL)法原位检测人肝癌(36例)和相关慢性肝病组织(慢性乙型肝炎、肝硬化和癌周肝硬化分别为20、20、36例)的p38MAPK、HBX、细胞周期G2/M期相关因子(cdc25B,p34cdc2,细胞周期蛋白B1)、细胞增殖因子Ki-67和细胞凋亡情况.根据资料不同分别采用x2检验、One-Way ANOVA或t检验进行统计学处理. 结果 HBX在慢性乙型肝炎(65.0％)和肝癌组(44.4％)阳性率较高,主要表达于胞核; p38MAPK、cdc25B、细胞周期蛋白B1、p34cdc2的阳性率从正常肝组织(40.0％,20.0％,20.0％,30.0％)、慢性乙型肝炎(60.0％,65.0％,40.0％,50.0％)、肝硬化(65.0％,75.0％,70.0％,55.0％)、癌周肝硬化(66.7％,75.0％,75.0％,63.9％)到肝癌组(77.8％,80.6％,80.6％,72.2％)逐渐增高,表达部位发生改变.p38MAPK胞核表达主要见于慢性乙型肝炎组和肝硬化组,胞质表达见于癌周肝硬化组和肝癌组,癌周肝硬化组与肝癌组之间的差异无统计学意义(x2=1.11,P＞0.05).正常肝、慢性乙型肝炎、肝硬化、癌周肝硬化和肝癌组织中的增殖指数(0.0000±0.000,0.0502±0.011,0.0411±0.009,0.0762±0.017,0.1810±0.036)和凋亡指数(0.0351±0.024,0.0607±0.022,0.0562±0.013,0.0716±0.011,0.1200±0.018)比较,除肝硬化外也逐渐增高,增殖指数在肝癌分化差组(0.2285±0.062)高于分化好组(0.1216±0.032,t=2.082,P=0.044),凋亡指数肝癌在分化好组(0.152±0.026)高于分化差组(0.081±0.022,t=2.129,P=0.041).结论肝癌发生过程中,HBX可能通过p38MAPK通路引起细胞周期、细胞增殖和凋亡的异常改变.细胞增殖基本与凋亡相伴随,肝癌增殖程度高于凋亡程度.癌周肝硬化不同于不伴癌的肝硬化,与肝癌的关系更密切.
목적 연구인간암발생과정중p38MAPK화을형간염병독X단백(HBX)대세포증식화조망적영향,탐토간암발생적궤제. 방법 응용면역조직화학화DNA원위말단표기(TUNEL)법원위검측인간암(36례)화상관만성간병조직(만성을형간염、간경화화암주간경화분별위20、20、36례)적p38MAPK、HBX、세포주기G2/M기상관인자(cdc25B,p34cdc2,세포주기단백B1)、세포증식인자Ki-67화세포조망정황.근거자료불동분별채용x2검험、One-Way ANOVA혹t검험진행통계학처리. 결과 HBX재만성을형간염(65.0％)화간암조(44.4％)양성솔교고,주요표체우포핵; p38MAPK、cdc25B、세포주기단백B1、p34cdc2적양성솔종정상간조직(40.0％,20.0％,20.0％,30.0％)、만성을형간염(60.0％,65.0％,40.0％,50.0％)、간경화(65.0％,75.0％,70.0％,55.0％)、암주간경화(66.7％,75.0％,75.0％,63.9％)도간암조(77.8％,80.6％,80.6％,72.2％)축점증고,표체부위발생개변.p38MAPK포핵표체주요견우만성을형간염조화간경화조,포질표체견우암주간경화조화간암조,암주간경화조여간암조지간적차이무통계학의의(x2=1.11,P＞0.05).정상간、만성을형간염、간경화、암주간경화화간암조직중적증식지수(0.0000±0.000,0.0502±0.011,0.0411±0.009,0.0762±0.017,0.1810±0.036)화조망지수(0.0351±0.024,0.0607±0.022,0.0562±0.013,0.0716±0.011,0.1200±0.018)비교,제간경화외야축점증고,증식지수재간암분화차조(0.2285±0.062)고우분화호조(0.1216±0.032,t=2.082,P=0.044),조망지수간암재분화호조(0.152±0.026)고우분화차조(0.081±0.022,t=2.129,P=0.041).결론 간암발생과정중,HBX가능통과p38MAPK통로인기세포주기、세포증식화조망적이상개변.세포증식기본여조망상반수,간암증식정도고우조망정도.암주간경화불동우불반암적간경화,여간암적관계경밀절.
Objective To investigate the effects of host-derived p38 mitogen-activated protein kinase subunit 38 (p38MAPK) and the hepatitis B virus X antigen (HbxAg) on cell proliferation and apoptosis in human hepatocellular carcinoma (HCC),and to study the mechanism underlying hepatocarcinogenesis.Methods Liver tissues were biopsied from healthy individuals and patients with chronic hepatitis B (CHB),liver cirrhosis,paratumor cirrhosis,and HCC.Immunohistochemical staining was used to detect expressions of HBxAg,p38MAPK,cell cycle G2/M phase-related factors (cdc25B,p34cdc2,cyclin B1),and cell proliferation factor ki-67.The terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling method (known as TUNEL) was used to detect apoptosis.Results The highest rates of HBxAg were detected in CHB (65.0％) and HCC (44.4％) liver samples,and the antigen was mainly expressed in nuclei.Increasingly higher rates of p38MAPK,cdc25B,cyclin B1,and p34cdc2 expression were detected with increases in disease severity:normal liver (40.0％,20.0％,20.0％,and 30.0％,respectively),chronic hepatitis B (60.0％,65.0％,40.0％,and 50.0％,respectively),liver cirrhosis (65.0％,75.0％,70.0％,and 55.0％,respectively),paratumor cirrhosis (66.7％,75.0％,75.0％,and 63.9％,respestively),and HCC (77.8％,80.6％,80.6％,and 72.2％,respectively).In addition,the intracellular location of p38MAPK expression was different under different disease conditions,showing nuclear expression in CHB and liver cirrhosis samples and cytoplasmic expression in paratumor cirrhosis and HCC samples (x2 =1.11,P ＞ 0.05).The proliferation index (PI) and the apoptosis index (AI) were both increased along with disease severity (nommal ＞ CHB ＞ paratumor cirrhosis ＞ HCC) (PI:0.0000 ± 0.000,0.0502 ± 0.011,0.0411 ± 0.009,0.0762 ± 0.017; AI:0.0351 ± 0.024,0.0607 ±0.022,0.0562 ± 0.013,0.0716 ± 0.011),with the notable exception for liver cirrhosis (PI:0.1810 ± 0.036 and AI:0.1200 ± 0.018).PI in poorly-differentiated HCC (0.2285 ± 0.062) was significantly higher than in welldifferentiated HCC (0.1216 ± 0.032,t =2.082,P =0.044).AI in well-differentiated HCC (0.152 ± 0.026) was significantly higher than in poorly-differentiated HCC (0.081 ± 0.022,t =2.129,P =0.041).Conclusions In the process of hepatocarcinogenesis,HBxAg may cause a series of abnormal changes in cell cycle,proliferation and apoptosis by affecting the expression of p38MAPK.