Synthetic artificial "long non-coding RNAs" targeting oncogenic microRNAs and transcriptional factors inhibit malignant phenotypes of bladder cancer cells

Haibiao Xie; Hengji Zhan; Qunjun Gao; Jianfa Li; Qun Zhou; Zhicong Chen; Yuhan Liu; Mengting Ding; Huizhong Xiao; Yuchen Liu; Weiren Huang; Zhiming Cai

doi:10.1016/j.canlet.2018.02.038

Synthetic artificial "long non-coding RNAs" targeting oncogenic microRNAs and transcriptional factors inhibit malignant phenotypes of bladder cancer cells

Cancer Lett. 2018 May 28:422:94-106. doi: 10.1016/j.canlet.2018.02.038. Epub 2018 Mar 1.

Authors

Haibiao Xie¹, Hengji Zhan², Qunjun Gao³, Jianfa Li⁴, Qun Zhou⁵, Zhicong Chen², Yuhan Liu², Mengting Ding², Huizhong Xiao², Yuchen Liu⁶, Weiren Huang⁷, Zhiming Cai⁸

Affiliations

¹ Key Laboratory of Medical Reprogramming Technology, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, 518039, China; Shantou University Medical College, Shantou, 515041, Guangdong Province, China; Department of Urology, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, 518039, China.
² Key Laboratory of Medical Reprogramming Technology, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, 518039, China; Department of Urology, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, 518039, China.
³ Key Laboratory of Medical Reprogramming Technology, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, 518039, China; Department of Urology, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, 518039, China; Guangzhou Medical University, Guangzhou, 511436, China.
⁴ Key Laboratory of Medical Reprogramming Technology, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, 518039, China; Shantou University Medical College, Shantou, 515041, Guangdong Province, China; Guangdong and Shenzhen Key Laboratory of Male Reproductive Medicine and Genetics, Institute of Urology, Peking University Shenzhen Hospital, Peking University, Shenzhen, 518036, China.
⁵ Key Laboratory of Medical Reprogramming Technology, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, 518039, China; Department of Urology, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, 518039, China; Shenzhen Second People's Hospital, Clinical Medicine College of Anhui Medical University, Shenzhen, 518039, Guangdong, China.
⁶ Key Laboratory of Medical Reprogramming Technology, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, 518039, China; Shantou University Medical College, Shantou, 515041, Guangdong Province, China. Electronic address: liuyuchenmdcg@163.com.
⁷ Key Laboratory of Medical Reprogramming Technology, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, 518039, China. Electronic address: pony8980@163.com.
⁸ Key Laboratory of Medical Reprogramming Technology, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, 518039, China; Shantou University Medical College, Shantou, 515041, Guangdong Province, China; Department of Urology, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, 518039, China. Electronic address: caizhiming2000@163.com.

PMID: 29501702
DOI: 10.1016/j.canlet.2018.02.038

Abstract

Both oncogenic transcription factors (TFs) and microRNAs (miRNAs) play important roles in human cancers, acting as transcriptional and post-transcriptional regulators, respectively. These phenomena raise questions about the ability of an artificial device to simultaneously regulate miRNAs and TFs. In this study, we aimed to construct artificial long non-coding RNAs, "alncRNAs", and to investigate their therapeutic effects on bladder cancer cell lines. Based on engineering principles of synthetic biology, we combined tandem arrayed aptamer cDNA sequences for TFs with tandem arrayed cDNA copies of binding sites for the miRNAs to construct alncRNAs. In order to prove the utility of this platform, we chose β-catenin and the miR-183-182-96 cluster as the functional targets and used the bladder cancer cell lines 5637 and SW780 as the test models. Dual-luciferase reporter assay, real-time quantitative PCR (qRT-PCR) and related phenotypic experiments were used to test the expression of related genes and the therapeutic effects of our devices. The result of dual-luciferase reporter assay and qRT-PCR showed that alncRNAs could inhibit transcriptional activity of TFs and expression of corresponding microRNAs. Using functional experiments, we observed decreased cell proliferation, increased apoptosis, and motility inhibition in alncRNA-infected bladder cancer cells. What's more, follow-up mechanism experiments further confirmed the anti-tumor effect of our devices. In summary, our synthetic devices indeed function as anti-tumor regulators, which synchronously accomplish transcriptional and post-transcriptional regulation in bladder cancer cells. Most importantly, anti-cancer effects were induced by the synthetic alncRNAs in the bladder cancer lines. Our devices, all in all, provided a novel strategy and methodology for cancer studies, and might show a great potential for cancer therapy if the challenges of in vivo DNA delivery are overcome.

Keywords: Aptamer; Bladder cancer; Synthetic biology; TFs; miRNA.

Publication types

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

MeSH terms

Aptamers, Nucleotide / biosynthesis
Aptamers, Nucleotide / pharmacology*
Cell Line, Tumor
Cell Movement / drug effects
Cell Proliferation / drug effects
Gene Expression Regulation, Neoplastic / drug effects
Humans
MicroRNAs / antagonists & inhibitors*
Phenotype
Promoter Regions, Genetic / drug effects
RNA, Long Noncoding / chemical synthesis
RNA, Long Noncoding / pharmacology*
Synthetic Biology
Urinary Bladder Neoplasms / drug therapy
Urinary Bladder Neoplasms / genetics*
beta Catenin / antagonists & inhibitors*

Substances

Aptamers, Nucleotide
CTNNB1 protein, human
MicroRNAs
RNA, Long Noncoding
beta Catenin