靶向反义长链非编码RNA HIF1A-AS2的四环素诱导型短发夹RNA抑制膀胱癌的恶性表型。

Tetracycline-inducible shRNA targeting antisense long non-coding RNA HIF1A-AS2 represses the malignant phenotypes of bladder cancer.

作者信息

Chen Mingwei, Zhuang Chengle, Liu Yuchen, Li Jianfa, Dai Fen, Xia Ming, Zhan Yonghao, Lin Junhao, Chen Zhicong, He Anbang, Xu Wen, Zhao Guoping, Guo Yinglu, Cai Zhiming, Huang Weiren

机构信息

Key Laboratory of Medical Reprogramming Technology, Shenzhen Second People's Hospital, Clinical Institute of Anhui Medical University, The First Affiliated Hospital of Shenzhen University, Shenzhen 518039, Guangdong Province, China; Department of Urology, The Fourth Affiliated Hospital of Zhejiang University School of Medicine, Yiwu 322000, Zhejiang Province, China; Anhui Medical University, Hefei 230032, Anhui Province, China.

Key Laboratory of Medical Reprogramming Technology, Shenzhen Second People's Hospital, Clinical Institute of Anhui Medical University, The First Affiliated Hospital of Shenzhen University, Shenzhen 518039, Guangdong Province, China; Shantou University Medical College, Shantou 515041, Guangdong Province, China.

出版信息

Cancer Lett. 2016 Jun 28;376(1):155-64. doi: 10.1016/j.canlet.2016.03.037. Epub 2016 Mar 24.

DOI:10.1016/j.canlet.2016.03.037

PMID:27018306

Abstract

Various studies have indicated that long non-coding RNAs (lncRNAs) play vital roles in the cancer development and progression. LncRNA hypoxia inducible factor 1alpha antisense RNA-2 (HIF1A-AS2) is upregulated in gastric carcinomas and knockdown of HIF1A-AS2 expression by siRNA could inhibit cell proliferation in vitro and tumorigenesis in vivo. Inspired by these observations, we hypothesized that HIF1A-AS2 possibly plays the analogous roles in bladder cancer. In our study, we first reported that HIF1A-AS2 was up-regulated in bladder cancer tissues and cells, and HIF1A-AS2 expression level in bladder cancer tissues is positively associated with advanced clinical pathologic grade and TNM phase. Cell proliferation inhibition, cell migration suppression and apoptosis induction were observed by silencing HIF1A-AS2 in bladder cancer T24 and 5637 cells. Overexpression of HIF1A-AS2 in SV-HUC-1 cells could promote cell proliferation, cell migration and anti-apoptosis. Besides, we utilized the emerging technology of medical synthetic biology to design tetracycline-inducible small hairpin RNA (shRNA) vector which specifically silenced HIF1A-AS2 in a dosage-dependent manner to inhibit the progression of human bladder cancer. In conclusion, our data suggested that HIF1A-AS2 plays oncogenic roles and can be used as a therapeutic target for treating human bladder cancer. Synthetic "tetracycline-on" switch system that quantitatively controlled the expression of HIF1A-AS2 in bladder cancer can inhibit the progression of bladder cancer cells in a dosage-dependent manner. Our findings provide new insights into the role of the lncRNA HIF1A-AS2 in the bladder cancer.

摘要

多项研究表明，长链非编码RNA（lncRNAs）在癌症的发生和发展中起着至关重要的作用。lncRNA缺氧诱导因子1α反义RNA-2（HIF1A-AS2）在胃癌中上调，通过小干扰RNA（siRNA）敲低HIF1A-AS2的表达可在体外抑制细胞增殖，并在体内抑制肿瘤发生。受这些观察结果的启发，我们推测HIF1A-AS2可能在膀胱癌中发挥类似作用。在我们的研究中，我们首先报道HIF1A-AS2在膀胱癌组织和细胞中上调，并且膀胱癌组织中的HIF1A-AS2表达水平与临床病理分级和TNM分期呈正相关。通过沉默膀胱癌T24和5637细胞中的HIF1A-AS2，观察到细胞增殖抑制、细胞迁移抑制和凋亡诱导。在SV-HUC-1细胞中过表达HIF1A-AS2可促进细胞增殖、细胞迁移和抗凋亡。此外，我们利用新兴的医学合成生物学技术设计了四环素诱导型小发夹RNA（shRNA）载体，该载体以剂量依赖性方式特异性沉默HIF1A-AS2，以抑制人膀胱癌的进展。总之，我们的数据表明HIF1A-AS2发挥致癌作用，可作为治疗人类膀胱癌的治疗靶点。定量控制膀胱癌中HIF1A-AS2表达的合成“四环素开启”开关系统可剂量依赖性地抑制膀胱癌细胞的进展。我们的研究结果为lncRNA HIF1A-AS2在膀胱癌中的作用提供了新的见解。