MiR-193a-3p and miR-193a-5p suppress the metastasis of human osteosarcoma cells by down-regulating Rab27B and SRR, respectively

Clinical & Experimental Metastasis, Feb 2016

MicroRNAs have been identified as key players in the development and progression of osteosarcoma, which is the most common primary malignancy of bone. Sequencing-based miR-omic and quantitative real-time PCR analyses suggested that the expression of miR-193a-3p and miR-193a-5p was decreased by DNA methylation at their promoter region in a highly metastatic osteosarcoma cell line (MG63.2) relative to their expression in the less metastatic MG63 cell line. Further wound-healing and invasion assays demonstrated that both miR-193a-3p and miR-193a-5p suppressed osteosarcoma cell migration and invasion. Moreover, introducing miR-193a-3p and miR-193a-5p mimics into MG63.2 cells or antagomiRs into MG63 cells confirmed their critical roles in osteosarcoma metastasis. Additionally, bioinformatics prediction along with biochemical assay results clearly suggested that the secretory small GTPase Rab27B and serine racemase (SRR) were direct targets of miR-193a-3p and miR-193a-5p, respectively. These two targets are indeed involved in the miR-193a-3p- and miR-193a-5p-induced suppression of osteosarcoma cell migration and invasion. MiR-193a-3p and miR-193a-5p play important roles in osteosarcoma metastasis through down-regulation of the Rab27B and SRR genes and therefore may serve as useful biomarkers for the diagnosis of osteosarcoma and as potential candidates for the treatment of metastatic osteosarcoma.

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MiR-193a-3p and miR-193a-5p suppress the metastasis of human osteosarcoma cells by down-regulating Rab27B and SRR, respectively

Clin Exp Metastasis MiR-193a-3p and miR-193a-5p suppress the metastasis of human osteosarcoma cells by down-regulating Rab27B and SRR, respectively Youguang Pu 0 1 2 3 4 Fangfang Zhao 0 1 2 3 4 Wenjing Cai 0 1 2 3 4 Xianghui Meng 0 1 2 3 4 Yinpeng Li 0 1 2 3 4 Shanbao Cai 0 1 2 3 4 0 Indiana University School of Medicine , Indianapolis, IN 46202 , USA 1 Cancer Epigenetics Program, Anhui Cancer Hospital , Hefei 230031, Anhui , China 2 & Shanbao Cai 3 Xinxiang Medical University , Xinxiang 453000, Henan , China 4 Department of Orthopedic Surgery, Anhui Cancer Hospital , Hefei 230031, Anhui , China MicroRNAs have been identified as key players in the development and progression of osteosarcoma, which is the most common primary malignancy of bone. Sequencing-based miR-omic and quantitative real-time PCR analyses suggested that the expression of miR-193a3p and miR-193a-5p was decreased by DNA methylation at their promoter region in a highly metastatic osteosarcoma cell line (MG63.2) relative to their expression in the less metastatic MG63 cell line. Further wound-healing and invasion assays demonstrated that both miR-193a-3p and miR-193a-5p suppressed osteosarcoma cell migration and invasion. Moreover, introducing miR-193a-3p and miR193a-5p mimics into MG63.2 cells or antagomiRs into MG63 cells confirmed their critical roles in osteosarcoma metastasis. Additionally, bioinformatics prediction along with biochemical assay results clearly suggested that the secretory small GTPase Rab27B and serine racemase (SRR) were direct targets of miR-193a-3p and miR-193a5p, respectively. These two targets are indeed involved in the miR-193a-3p- and miR-193a-5p-induced suppression of osteosarcoma cell migration and invasion. MiR-193a-3p and miR-193a-5p play important roles in osteosarcoma metastasis through down-regulation of the Rab27B and SRR genes and therefore may serve as useful biomarkers for the diagnosis of osteosarcoma and as potential candidates for the treatment of metastatic osteosarcoma. Rab27B; SRR; MiR-193a-3p and miR-193a-; 5p; Osteosarcoma; Metastasis - Youguang Pu and Fangfang Zhao have contributed equally to this work. Introduction Osteosarcoma is the most common primary bone malignancy in children and young adults [ 1, 2 ]. Approximately 80 % of osteosarcoma patients have metastatic disease at the time of diagnosis, and metastasis is a consistent problem in tumor prognosis and treatment [3]. The molecular mechanisms that lead to the development and metastasis of osteosarcoma are poorly understood [ 4–6 ]. Further exploration of this area will help in the development of effective strategies in the diagnosis, treatment and prognosis of osteosarcoma. Previous studies have established a highly tumorigenic and metastatic human osteosarcoma cell line, MG63.2, derived from the less metastatic parental MG63 cell line [ 7 ]. The MG63.2 cell line facilitates the exploration of crucial players in osteosarcoma metastasis [ 8 ]. MicroRNAs (miRNAs), a class of small non-coding RNA molecules, play critical roles in a variety of biological events, including development, cell proliferation and cell differentiation [ 9–11 ]. MiRNAs negatively regulate gene expression by binding to the 30-untranslated regions (UTRs) of the corresponding target mRNAs of proteincoding genes, thereby leading to mRNA degradation or translation inhibition [ 12–15 ]. Multiple miRNAs are involved in the invasion and metastasis of different types of cancers, including gastric cancer, breast cancer, hepatocellular carcinoma and colorectalcancer [ 16–19 ]. In osteosarcoma, a few miRNAs have been reported to regulate metastasis. For instance, miR-16 inhibits cell proliferation by targeting IGF1R and the Raf1-MEK1/2-ERK1/2 pathway in osteosarcoma [ 20 ], and miR-802 promotes osteosarcoma cell proliferation by down-regulating the p27 cell-cycle inhibitor [ 21 ]. Moreover, miR-212 inhibits osteosarcoma cell proliferation and invasion by downregulating Sox4 [ 22 ]. However, the prospects of these miRNAs in the clinical treatment of osteosarcoma have not been evaluated. Novel miRNAs that may potentially serve as new therapeutic targets or biomarkers must urgently be identified. In the present study, we aimed to identify novel miRNAs associated with osteosarcoma metastasis. From a sequencing-based miR-omic study and referred to the relevant literatures, we found two miRNAs, miR-193a-3p and miR-193a-5p, that play a role in osteosarcoma metastasis. These two miRNAs, which are derived from the same precursor, have different sequences and therefore target different mRNAs [ 23 ]. These miRNAs have been implicated in the tumorigenesis and progression of several types of cancer [ 24–28 ]. In addition, we identified the target genes of miR-193a-3p and miR-193a-5p, and we proposed the potential pathway that might be associated with osteosarcoma metastasis. Therefore, miR-193a-3p and miR-193a-5p, as well as their targets, might serve as (...truncated)


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Youguang Pu, Fangfang Zhao, Wenjing Cai, Xianghui Meng, Yinpeng Li, Shanbao Cai. MiR-193a-3p and miR-193a-5p suppress the metastasis of human osteosarcoma cells by down-regulating Rab27B and SRR, respectively, Clinical & Experimental Metastasis, 2016, pp. 359-372, Volume 33, Issue 4, DOI: 10.1007/s10585-016-9783-0