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Development of synthetic microRNA‐214 showing enhanced cytotoxicity and RNase resistance for treatment of canine hemangiosarcoma
Author(s) -
Yoshikawa Ryutaro,
Heishima Kazuki,
Ueno Yoshihito,
Kawade Miwa,
Maeda Yusuke,
Yoshida Kyoko,
Murakami Mami,
Sakai Hiroki,
Akao Yukihiro,
Mori Takashi
Publication year - 2020
Publication title -
veterinary and comparative oncology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.864
H-Index - 34
eISSN - 1476-5829
pISSN - 1476-5810
DOI - 10.1111/vco.12580
Subject(s) - transfection , microrna , cytotoxicity , in vivo , rnase p , nuclease , in vitro , gene silencing , antagomir , chemistry , apoptosis , cancer research , cell culture , microbiology and biotechnology , biology , rna , biochemistry , gene , genetics
MicroRNA‐214 (miR‐214), a pivotal tumour‐suppressive miRNA, is downregulated in canine hemangiosarcoma (HSA) cells. Although these tumour‐suppressive miRNAs are potential therapeutic agents, their clinical efficacy may be limited because of their vulnerability to RNase‐rich microenvironments and low in vivo transfection rates. We developed synthetic miR‐214s with enhanced cytotoxicity, RNase resistance and quantity of miR‐214 in/on cells. These synthetic miR‐214s were synthesized by various chemical modifications (such as 4′‐aminoethyl‐2′‐fluoro, 2′‐fluoro, 2′‐O‐methyl, phosphorothioate and oligospermine modifications) of the wild‐type mature miR‐214 sequences. Transfection of HSA cells with synthetic miR‐214 (miR‐214 5AE) demonstrated significant growth suppressive effect and induced the strongest apoptotic response. Synthetic miR‐214s (miR‐214 5AE, miR‐214 10AE and miR‐214 OS) were much more stable than mature miR‐214s in foetal bovine serum. Similar to mature miR‐214, 5AE and OS suppressed the expression level of COP1 in HSA cells. The quantity of synthetic miR‐214s in/on cells was higher than that of mature miR‐214. In conclusion, we developed a clinically applicable, synthetic miR‐214 5AE that regulates the COP1 protein expression similar to that mediated by mature miR‐214. Additionally, miR‐214 5AE confers better cytotoxicity, nuclease resistance and transfection rate than mature miR‐214. Thus, miR‐214 5AE could potentially be a novel miRNA‐based chemotherapeutic agent that could improve the prognosis of HSA. Its in vivo effects on canine HSA need to be examined in future.