过表达SIRT3抑制裸鼠异位移植人肝细胞癌的生长及其机制研究  

Overexpression of SIRT3 Inhibits Human Hepatocellular Carcinoma Growth in Ectopic Xenograft Nude Mouse Model and Its Mechanism

作　　者：陈祥[1] 任吉华[1] 冉龙宽 陶娜娜[1] 李宛蔚[1] 周洪钟[1] 刘波[1] 陈娟[1] 

机构地区：[1]重庆医科大学感染性疾病分子生物学教育部重点实验室,重庆400016

出　　处：《中国细胞生物学学报》2015年第12期1653-1658,共6页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81472271;81201282)资助的课题~~

摘　　要：该文旨在研究人肝细胞癌异位移植瘤裸鼠模型中沉默信息调节因子3(silent information regulator 3,SIRT3)对肝细胞癌生长的影响及其机制。建立稳定过表达SIRT3和pc DNA3.1的SK-Hep-1细胞株;将稳定过表达SIRT3和pc DNA3.1的细胞悬液分别注射入裸鼠皮下,实时监测两组移植瘤的生长,25 d后剥离出移植瘤并称重;免疫组织化学检测移植瘤中SIRT3、Ki67的表达水平;应用定量逆转录PCR(q RT-PCR)筛选SIRT3影响移植瘤生长的下游靶向分子,Western blot检测下游靶向分子Bax的表达量以及移植瘤中cleaved-PARP(poly ADP-ribose polymerase)的表达水平。结果显示,过表达SIRT3组移植瘤的体积和重量都小于pc DNA3.1组;过表达SIRT3组移植瘤中Ki67的表达水平较pc DNA3.1组降低;过表达SIRT3上调Bax的m RNA和蛋白质水平并促进PARP的剪切。该文结果提示,SIRT3可能通过Bax凋亡信号通路抑制人肝细胞癌异位移植瘤的生长。This study investigated the effect of silent information regulator 3 （SIRT3） on the growth of HCC in ectopic xenograft nude mouse model and its possible molecular mechanism. SK-Hep-1 cells were es- tablished with stable overexpression of SIRT3 or pcDNA3.1, respectively. Then, these cells were injected to the subcutaneous of nude mice, respectively. The growth of ectopic xenografl in SIRT3 group and pcDNA3.1 group were monitored at regular intervals. The ectopic xenograft were stripped out and weighed at 25th day after injec- tion. Expression levels of SIRT3 and Ki-67 were determined by immunohistoehemieal staining. Downstream target molecules, which may participate in the SIRT3 pathway that regulated the growth of transplantation tumor, were analyzed by reverse-transcription polymerase chain reaction （qRT-PCR）. Bax and cleaved-PARP were detected by Western blot. The results showed that SIRT3 overexpression could decrease the size and weight of ectopic xeno- graft and downregulate the expression of Ki-67. Furthermore, overexpression of SIRT3 raised the Bax in mRNA and protein levels, and promoted the protein cleavage of PARP. These results demonstrated that SIRT3 might inhibitthe growth of human HCC ectopic xenografl by Bax signaling pathway in apoptosis.

关 键 词：肝细胞癌 移植瘤 SIRT3 BAX 生长 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤]