CAJ | 학술논문

目的研究乙肝病毒/黄曲霉素B1相关的肝细胞癌(HCC)差异蛋白质表达谱特点,初步探讨两因素的协同致癌机制.方法 32例HCC患者的癌组织按照乙肝病毒与黄曲霉毒素B1的暴露情况分为四个亚组:组A:H BV(+)/AFB1(+)10例;组B:HBV(+)/AFB1(-)10例;组C:HBV(-)/AFB1(+)6例;组D:HBV(-)/AFB1(-)6例.收集10例来自肝血管瘤、肝外伤、肝移植供者等手术切除的正常肝组织作为对照.应用iTRAQ结合2DLC-MS/MS技术分析蛋白质表达谱在4个亚组间的差异.结果 (1)共鉴定出117种差异蛋白,包括53种上调蛋白和64种下调蛋白.四个亚组中上调和下调的差异蛋白分别为组A 106种,组B 97种,组C 104种,组D 74种.(2)117种差异蛋白中,有9种为热休克家族蛋白(HSPs),它们上调表达在组A、组B与组C中.有15种蛋白为毒物代谢解毒相关酶蛋白,其中的12种下调表达在组A中,且半数以上也下调表达在组B与组C中.(3)对差异蛋白AKR1B10的验证结果显示:AKR1B10基因mRNA在组A中的表达分别与组B、组C以及组D相比,差异具有统计学意义(均P＜0.05).此外,组C与组D相比,差异也具有统计学意义(P＜0.05);AKR1B10蛋白在组A中的表达与组B、组D相比,差异具有统计学意义(均P＜0.05).结论 HSPs的上调表达及毒物代谢酶相关蛋白的下调表达为乙肝病毒和黄曲霉素B1暴露相关的肝细胞癌常见的分子生物学事件.研究提示乙肝病毒/黄曲霉素B1双因素的协同致癌机制,可能与两者导致毒物代谢酶相关蛋白失调有关.AKR1B10的高表达可能参与了AFB1的致肝癌过程.
목적 연구을간병독/황곡매소B1상관적간세포암(HCC)차이단백질표체보특점,초보탐토량인소적협동치암궤제.방법 32례HCC환자적암조직안조을간병독여황곡매독소B1적폭로정황분위사개아조:조A:H BV(+)/AFB1(+)10례;조B:HBV(+)/AFB1(-)10례;조C:HBV(-)/AFB1(+)6례;조D:HBV(-)/AFB1(-)6례.수집10례래자간혈관류、간외상、간이식공자등수술절제적정상간조직작위대조.응용iTRAQ결합2DLC-MS/MS기술분석단백질표체보재4개아조간적차이.결과 (1)공감정출117충차이단백,포괄53충상조단백화64충하조단백.사개아조중상조화하조적차이단백분별위조A 106충,조B 97충,조C 104충,조D 74충.(2)117충차이단백중,유9충위열휴극가족단백(HSPs),타문상조표체재조A、조B여조C중.유15충단백위독물대사해독상관매단백,기중적12충하조표체재조A중,차반수이상야하조표체재조B여조C중.(3)대차이단백AKR1B10적험증결과현시:AKR1B10기인mRNA재조A중적표체분별여조B、조C이급조D상비,차이구유통계학의의(균P＜0.05).차외,조C여조D상비,차이야구유통계학의의(P＜0.05);AKR1B10단백재조A중적표체여조B、조D상비,차이구유통계학의의(균P＜0.05).결론 HSPs적상조표체급독물대사매상관단백적하조표체위을간병독화황곡매소B1폭로상관적간세포암상견적분자생물학사건.연구제시을간병독/황곡매소B1쌍인소적협동치암궤제,가능여량자도치독물대사매상관단백실조유관.AKR1B10적고표체가능삼여료AFB1적치간암과정.
Objective To explore the effects and the molecular mechanism of synergistic hepatocarcinogenesis of HBV and AFB1 in the development of HCC by studying the difference in protein expression profiles in hepatocellular carcinoma exposed to hepatitis B virus and Aflatoxin B1.Method 32 HCC specimens were labeled under four categories based on their biomarkers of HBV and AFB1 exposure:group A:HBV(+)/AFB1 (+),10 cases,group B:HBV(+)/AFB1 (-),10 cases,group C:HBV(-)/AFB1(+),6 cases,groupD:HBV(-)/AFB1(-),6 cases.Normal hepatic tissues from 10 cases of hepatic hemangioma,liver resection and liver transplant donor were chosen as the normal control group.Isobaric Tagging Reagent Quantitative (iTRAQ) Proteomics together with 2DLC MS/MS were applied to analyze the differentially expressed proteins among the 4 groups.Result (1) A total of 117 unique differentially expressed proteins including 53 up-regulated proteins and 64 down-regulated proteins were identified in the four groups.The number of unique differentially expressed proteins,including up-regulated and down-regulated proteins,in group A,group B,group C and group D were 106,97,104 and 74 respectively.(2) Among the 117 differentially expressed proteins,9 proteins were heat shock proteins or chaperones,and they were up regulated in group A,B and C.Besides,15 proteins were detoxification and drug metabolism pathway related proteins,12 of them were down-regulated in group A,and more than half of them were also down-regulated in group B and C.(3) The Reverse Transcriptase PCR result showed the mean expression level of AKR1B10 mRNA in group A was significantly higher than group B,C and D (all P＜0.05,respectively).Group C also showed significantly a higher expression level of AKR1B10 mRNA than group D (P＜0.05).The Western-blot results showed the mean expression level of AKR1B10 protein in group A was significantly higher than group B and D (all P＜0.05,respectively).Conclusions The up-regulated expression of heat shock protein and the down-regulated expression of most protein enzymes related to detoxification and drug metabolism were the common molecular biological events of HCC associated with exposure to HBV and AFB1.This suggested the synergistic hepatocarcinogenesis effects of HBV and AFB1 may be related to dysregulation of protein enzymes related to detoxification and drug metabolism.The overexpression of AKR1B10 may be involved in the AFB1-related hepatocarcinogenesis process.