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

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

doi:10.1016/j.canlet.2016.03.037

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

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

Authors

Mingwei Chen¹, Chengle Zhuang², Yuchen Liu³, Jianfa Li², Fen Dai⁴, Ming Xia⁵, Yonghao Zhan², Junhao Lin², Zhicong Chen², Anbang He⁶, Wen Xu³, Guoping Zhao⁷, Yinglu Guo⁸, Zhiming Cai⁹, Weiren Huang¹⁰

Affiliations

¹ 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.
² 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.
³ 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.
⁴ Anhui Medical University, Hefei 230032, Anhui Province, China.
⁵ Department of Urology, The Third Affiliated Hospital of Southen Medical University, Guangzhou 510630, Guangdong Province, China.
⁶ 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; Anhui Medical University, Hefei 230032, Anhui Province, China.
⁷ 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; Shanghai-MOST Key Laboratory of Health and Disease Genomics, Chinese National Human Genome Center at Shanghai, Shanghai 200000, China.
⁸ Department of Urology, Peking University First Hospital, Institute of Urology, Peking University, National Urological Cancer Center, Beijing 100034, China.
⁹ 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; Anhui Medical University, Hefei 230032, Anhui Province, China; Shantou University Medical College, Shantou 515041, Guangdong Province, China; Department of Urology, Peking University First Hospital, Institute of Urology, Peking University, National Urological Cancer Center, Beijing 100034, China. Electronic address: caizhiming2000@163.com.
¹⁰ 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; Anhui Medical University, Hefei 230032, Anhui Province, China; Shantou University Medical College, Shantou 515041, Guangdong Province, China; Department of Urology, Peking University First Hospital, Institute of Urology, Peking University, National Urological Cancer Center, Beijing 100034, China. Electronic address: pony8980@163.com.

PMID: 27018306
DOI: 10.1016/j.canlet.2016.03.037

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.

Keywords: Bladder cancer; HIF1A-AS2; LncRNAs; Synthetic biology; Tetracycline-inducible.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Apoptosis
Cell Line, Tumor
Cell Movement
Cell Proliferation
Female
Gene Expression Regulation, Neoplastic / drug effects*
Humans
Male
Middle Aged
Neoplasm Invasiveness
Phenotype
RNA Interference*
RNA, Long Noncoding / genetics*
RNA, Long Noncoding / metabolism
RNA, Small Interfering / biosynthesis
RNA, Small Interfering / genetics*
RNAi Therapeutics / methods*
Tetracycline / pharmacology*
Time Factors
Transfection
Up-Regulation
Urinary Bladder Neoplasms / genetics
Urinary Bladder Neoplasms / metabolism
Urinary Bladder Neoplasms / pathology
Urinary Bladder Neoplasms / therapy*

Substances

RNA, Long Noncoding
RNA, Small Interfering
long noncoding RNA HIF1A-AS2, human
Tetracycline