摘要
目的研究5-氟尿嘧啶(5-FU)对miR-22在肝癌细胞中表达的影响及其抑制肝癌细胞增殖的分子机制。方法用Real-time PCR法检测肝细胞癌组织及细胞系中miR-22的表达;用Real-Time PCR法检测经不同浓度5-FU处理的肝癌细胞HepG2和Huh7中miR-22及其初始转录产物pri-miR-22的表达;用miR-22类似物及5-FU处理肝癌细胞HepG2和Huh7,并用Western blot法检测miR-22靶基因HDAC4的表达;设计拯救实验(rescue assay)研究5-FU、miR-22与肝癌细胞增殖的关系。结果 miR-22在肝细胞癌组织及细胞系中表达下调;5-FU可以诱导肝癌细胞系HepG2、Huh7内源性miR-22、pri-miR-22表达水平明显上调(P<0.01);miR-22过表达及5-FU处理可以抑制HepG2、Huh7,HDAC4蛋白表达水平(P<0.01)。结论 5-FU通过调控miR-22介导的HDAC4信号通路发挥抑制肝癌细胞增殖的作用。
Objective To investigate the impact of 5-fluorouracil( 5-FU) on miR-22 expression in human hepatocellular carcinoma( HCC) cell lines and to elucidate the molecular mechanism of 5-fluorouracil for HCC chemotherapy. Methods Real-time PCR analysis was conducted to determine the expression levels of miR-22 in HCC tissue specimens and HCC cell lines. The expression of miR-22 and pri-miR-22( primary miR-22) was evaluated in HepG2 and Huh7 cells with 5-FU treatment by using real-time PCR and we also performed Western blot analysis to detect the protein level of HDAC4 in HCC cells with the same treatment. A rescue assay was employed by using5-FU treatment in combination with miR-22 inhibitor( Anti-22) to further investigate the correlation among 5-FU,miR-22,and HCC cell growth. Results miR-22 expression depicted a significant downregulation in HCC tissues and cell lines( P〈0. 01). 5-FU treatment led to an augment of miR-22 expression in HepG2 and Huh7 cells( P〈0. 001) and resulted in a decrease of HDAC4 protein levels,which was verified as a direct target of miR-22 in HCC cells( P〈0. 01). Conclusions 5-FU has suppressive effect on HCC growth which could be potentially explained by miR-22-mediated HDAC4 axis.
作者
孙蒙清
杜永星
李宗泽
由磊
舒红
刘子文
SUN Meng-qing DU Yong-xing LI Zong-ze YOU Lei SHU Hong LIU Zi-wen(Dept. of General Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Beijing 100730, China)
出处
《基础医学与临床》
CSCD
2017年第10期1368-1372,共5页
Basic and Clinical Medicine
基金
国家自然科学基金(81272767)