Guan Jinping, Liu Zimin, Xiao Menjing, Hao Fengyun, Wang Chenghong, Chen Yan, Lu Yingying, Liang Jun
Department of General Surgery, The Affiliated Hospital of Qingdao UniversityQingdao, Shandong, China.
Department of Oncology, The Affiliated Hospital of Qingdao UniversityQingdao, Shandong, China.
Am J Transl Res. 2017 May 15;9(5):2457-2465. eCollection 2017.
MicroRNAs play an important role in cell proliferation, apoptosis, differentiation, and invasion by regulating the expression of various genes. For example, the downregulation of microRNA-199a-3p (miR-199a-3p) that is noted in numerous human malignancies, including hepatocellular carcinoma (HCC), results in a poor prognosis in patients with HCC. This finding suggests that miR-199a-3p overexpression in HCC could provide a new treatment approach. We explored this possibility by examining the effects of miR-199a-3p on the growth and apoptosis of HCC cells in vitro and vivo.
The miR-199a-3p signaling pathway was examined using ZHX1 (zinc-fingers and homeoboxes-1) or PUMA (a p53 upregulated modulator of apoptosis) siRNA transfection to determine the effects of miR-199a-3p on growth and apoptosis of HepG2 cells in vitro. A subcutaneously implanted tumor model of HepG2 cells in nude mice was used to assess the effects of miR-199a-3p on the signaling pathway and tumorigenesis development in vivo.
miR-199a-3p inhibited growth and induced apoptosis of HepG2 cells in vitro. These effects were accompanied by upregulation of ZHX1 and PUMA. Targeting ZHX1 inhibited upregulation of PUMA after miR-199a-3p transfection. In addition, miR-199a-3p inhibited Bcl2 expression, but increased Bax and cleaved caspase-3 expression. Targeting PUMA or ZHX1 reversed the effect of miR-199a-3p, followed by upregulation of Bcl2 and downregulation of Bax and cleaved caspase-3, respectively. Furthermore, miR-199a-3p inhibited tumorigenesis of xenografts in nude mice.
miRNA-199a-3p could effectively prevent primary tumor formation. The ability of this therapy to decrease tumorigenesis may be related toZHX1-dependent PUMA signals.
微小RNA通过调控多种基因的表达，在细胞增殖、凋亡、分化及侵袭过程中发挥重要作用。例如，在包括肝细胞癌（HCC）在内的多种人类恶性肿瘤中均发现微小RNA - 199a - 3p（miR - 199a - 3p）表达下调，这导致HCC患者预后不良。该发现提示，在HCC中过表达miR - 199a - 3p可能提供一种新的治疗方法。我们通过检测miR - 199a - 3p对体外及体内HCC细胞生长和凋亡的影响来探索这种可能性。
使用锌指和同源盒蛋白1（ZHX1）或p53上调凋亡调节因子（PUMA）的小干扰RNA（siRNA）转染来检测miR - 199a - 3p信号通路，以确定miR - 199a - 3p对体外HepG2细胞生长和凋亡的影响。利用裸鼠皮下植入HepG2细胞的肿瘤模型来评估miR - 199a - 3p对体内信号通路及肿瘤发生发展的影响。
miR - 199a - 3p在体外抑制HepG2细胞生长并诱导其凋亡。这些作用伴随着ZHX1和PUMA的上调。靶向ZHX1可抑制miR - 199a - 3p转染后PUMA的上调。此外，miR - 199a - 3p抑制Bcl2表达，但增加Bax和裂解的caspase - 3表达。靶向PUMA或ZHX1可逆转miR - 199a - 3p的作用，随后分别上调Bcl2以及下调Bax和裂解的caspase - 3。此外，miR - 199a - 3p抑制裸鼠异种移植瘤的发生。
微小RNA - 199a - 3p可有效预防原发性肿瘤形成。该疗法降低肿瘤发生的能力可能与ZHX1依赖的PUMA信号有关。
