Guangdong and Shenzhen Key Laboratory of Male Reproductive Medicine and Genetics, Institute of Urology, Peking University Shenzhen Hospital, Shenzhen-Peking University-the Hong Kong University of Science and Technology Medical Center, Shenzhen, 518000, People's Republic of China.
The Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, 100191, People's Republic of China.
J Exp Clin Cancer Res. 2017 Oct 30;36(1):151. doi: 10.1186/s13046-017-0626-x.
The traditional treatment for cancer is lack of specificity and efficacy. Modular synthetic regulatory RNAs, such as inhibitive RNA (iRNA) and active RNA (aRNA), may overcome these limitations. Here, we synthesize a new iRNA to bind the upstream activating sequence (UAS) of a minimal promoter that drives expression of artificial miRNAs (amiRNAs) targeting MYC, which represses the binding interaction between UAS and GAL4 fusion protein (GAL4-VP64) in GAL4/UAS system. The aRNA driven by a tumor-specific mutant human telomerase reverse transcriptase (hTERT) promoter is created to interact with iRNA to expose UAS again in bladder cancer. Without the aRNA, mRNA and protein levels of MYC, cell growth, cell apoptosis and cell migration were no significance in two bladder cancer cell lines, T24 and 5637, and human foreskin fibroblast (HFF) cells. The aRNA significantly inhibited the expression of MYC in mRNA and protein levels, as well as the proliferation and migration of the cancer cells, but not in HFF cells. These results indicated that regulatory RNAs selectively controlled the expression of amiRNAs and ultimately suppress the progression of bladder cancer cells without affecting normal cells. Synthetic regulatory RNAs might be a selective therapeutic approach for bladder cancer.
传统的癌症治疗方法缺乏特异性和疗效。模块化合成调控 RNA,如抑制性 RNA(iRNA)和激活性 RNA(aRNA),可能克服这些限制。在这里,我们合成了一种新的 iRNA,以结合最小启动子的上游激活序列(UAS),该启动子驱动靶向 MYC 的人工 miRNA(amiRNA)的表达,从而抑制 GAL4/UAS 系统中 UAS 与 GAL4 融合蛋白(GAL4-VP64)的结合相互作用。由肿瘤特异性突变型人端粒酶逆转录酶(hTERT)启动子驱动的 aRNA 被创建为与 iRNA 相互作用,以在膀胱癌中再次暴露 UAS。在没有 aRNA 的情况下,膀胱癌细胞系 T24 和 5637 以及人包皮成纤维细胞(HFF)细胞中 MYC 的 mRNA 和蛋白水平、细胞生长、细胞凋亡和细胞迁移均无显著差异。aRNA 显著抑制了 MYC 的表达,包括 mRNA 和蛋白水平的表达,以及癌细胞的增殖和迁移,但对 HFF 细胞没有影响。这些结果表明,调控 RNA 可以选择性地控制 amiRNA 的表达,最终抑制膀胱癌细胞的进展,而不影响正常细胞。合成调控 RNA 可能是膀胱癌的一种选择性治疗方法。