饥饿诱导DRAM介导的自噬与肝细胞凋亡的关系  

The relationship between DRAM-mediated autophagy and apoptosis in liver cells in response to starvation

作　　者：刘凯[1] 姜涛[1] 王安娜[1] 温韬[1] 殷继明[1] 任峰[1] 时红波[1] 刘道洁[1] 乔录新[1] 李宁[1] 陈德喜[1] 

机构地区：[1]首都医科大学附属北京佑安医院北京市肝病研究所,100069

出　　处：《北京医学》2013年第6期434-436,I0003,共4页Beijing Medical Journal

基　　金：国家自然科学基金(81071843);北京市卫生系统高层次卫生技术人才培养计划;北京市肝病研究所2012年度双十计划(NO.Z121107002812113);北京市肝病研究所所内基金(BJIH-01201)

摘　　要：目的探讨在饥饿情况下,自噬与正常肝脏细胞凋亡的关系,及凋亡与损伤调节自噬调控基因(dam-age-regulated autophagy modulator,DRAM)介导的自噬之间的关系。方法采用无血清培养基培养正常肝脏细胞系7702细胞36h,采用M30免疫荧光染色观察凋亡,采用免疫印记检测自噬和DRAM基因的表达水平;并用DRAM特异的小干扰RNA(siRNA)阻断DRAM作用后,观察凋亡和自噬的水平。结果和0h相比,在12、24和36h3个时间点,饥饿可明显诱导7702细胞凋亡(P<0.001)。免疫印记结果显示,饥饿后第12、24和36小时的DRAM水平和自噬水平均逐渐增高,两两比较,差异有统计学意义(P<0.05)。用DRAMsiRNA阻断DRAM功能后,在12、24和36h未检测到DRAM的表达,同时凋亡细胞数明显下降(P<0.05)。DRAMsiRNA阻断DRAM功能后,在12、24和36h的自噬水平无明显差异。结论 DRAM介导的自噬可导致正常肝脏细胞凋亡,其对凋亡的影响可能成为肝癌研究的新思路。Objective To study the relationship between autophagy and apoptosis in normal liver cells, and the relationship between apoptosis and damage-regulated autophagy modulator（DRAM） mediated autophagy and apoptosis in liver cells in response to starvation. Methods Normal liver cells 7702 were cultured in serum-free culture medium for 36 h, M30 immunofluorescence staining was used to assess apoptosis, immune imprint was used to detect the expression of au- tophagy and DRAM gene, and observed the level of apoptosis and autophagy after occlusion DRAM effects by DRAM-spe- cific siRNA. Results At 12, 24 and 36 h time points, nutritional deprivation induced apoptosis of 7702 cells significantly, compare with the time point of 0 h. Immune imprinting results suggested that the expression of DRAM induced by nutri- tional deprivation and autophagy gradually increased with extension of time, and there were statistical differences. At 12, 24 and 36 h time points, the expression of DRAM was not detected for occlusion DRAM function by DRAM-specific siRNA, and meanwhile the number of apoptotic cells decreased significantly（P 〈 0.05）. By occlusion DRAM function using DRAM-specific siRNA, autophagy had no significant differences among 12, 24 and 36 h time points. It indicated that DRAM mediated autophagy was a part of autophagy induced by nutritional deprivation. Conclusion DRAM mediated autophagy could lead to apoptosis of normal liver cells. It may be one of the mechanisms that normal liver cells occur apoptosis actively and prevent formating carcinoma when serum deprivation or blood supply depression.

关 键 词：损伤调节自噬调控基因 凋亡 自噬 

分 类 号：R363[医药卫生—病理学]