诱导肝细胞L02和肝癌细胞MHCC97L产生的氧化应激与Yes相关蛋白的关系  

Oxidative stress of L02 cells and MHCC97L cells and its correlation with Yes - associated protein

作　　者：乔欣[1] 李华[1] 吴小才 朱曙光[1] 杨扬[1] 陈规划[1] 

机构地区：[1]中山大学附属第三医院肝移植中心中山大学器官移植研究所广东省器官移植研究中心广东省肝脏疾病研究重点实验室,广州510630

出　　处：《中华实验外科杂志》2015年第8期1876-1878,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81172038、81172036、81170451、81370575、81370555）;国家十二五重大科技专项（2012ZX10002017-005、2012ZX10002016-011、2012ZX10002010-001-007）

摘　　要：目的通过过氧化氢诱导肝细胞及肝癌细胞产生氧化应激，探讨氧化应激与Hippo信号通路中核心蛋白Yes相关蛋白（YAP）的相关性。方法利用过氧化氢诱导构建氧化应激模型，用200μmol／L过氧化氢作用于正常人肝细胞L02与肝癌细胞MHCC97L6h，Westernblot检测YAP蛋白表达的差异，反转录-聚合酶链反应（1IT—PCR）检测YAP基因mRNA表达差异，流式细胞术检测L02细胞周期的变化，比较实验组和对照组差异。结果200μmol／L的过氧化氢作用于L02细胞及MHCC97L细胞6h后，与对照组比较，实验组两种细胞YAP蛋白表达明显升高。L02细胞中YAP基因mRNA表达水平上调479．5％，差异有统计学意义（P〈0．01），MHCC97L细胞上调4．7％，蓐肄尤统计学意义（P〉0．05）。流式细胞术检测细胞周期，L02细胞实验组与对照组比较s期细胞比例娃著升高[（46．40±4．88）％比（28．61±1．01）％]，差异有统计学意义（P〈0．01），G1期[（45．21±9．45）％比（59．59±2．44）％和G2期（8．42±5．14）％比（11．70±3．35）％]细胞分布改变不明显，差异无统计学意义（P〉0．05）。结论在氧化应激状态的L02细胞及MHCC97L细胞中，Itippo信号通路存在调控紊乱，核心蛋白YAP表达上调。其中1．02细胞存在转录水平的调控机制，MHCC97L细胞存在转录后水平的调控机制。氧化应激可以促进L02细胞增殖，YAP蛋白的表达升高可能是其中的原因之一。Objective To investigate the correlation between H202 - induced oxidative stress of 1122 cells and MHCC971, cells with Yes - associated protein （YAP） expression. Methods L02 cells and MHCC97L cells were exposed to 200 μmol/L H2O2 to establish the oxidative stress model. The expression of YAP protein and mRNA was detected by Western blotting and reverse transcriptase -polymerase chain re, action （ RT - PCR） , respectively. The cell cycles were examined by flow cytometry. Results After trea- ted with 200 μmol/L H202 for 6 h, the expression of YAP protein was up - regulated in both L02 cells and MHCC97L cells as compared with the control group. The reverse transcriptase - PCR results showed that the YAP mRNA levels were significant increased by 479. 5% （P 〈0.01 ） in L02 cells but had no signifi- ,ant change （ increased by 4. 7% , P 〉 0. 05 ） in MHCC97L cells. The flow eytometry results indicated more 1122 cells went into S phase [ （46. 40 ±4. 88） % vs. （28.61 ± 1.01 ） % ,P 〈0. 01 ] and ther were no sig- nificant changes in G1 , and G2 phases [ （45.21 ± 9.45 ）% vs. （59. 59 ± 2.44） % , and （8.42 ± 5. 14）% vs. （ 11.70 ±3.35 ） % , P 〉 0. 05 ]. Conclusion Oxidative stress can promote YAP protein expression in I D2 cells and MHCC97L cells, and promote proliferation of LO2 cells.

关 键 词：过氧化氢 氧化应激 Yes相关蛋白 细胞周期 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]