目的:探讨miR-362-3p对OS的影响及其相关信号通路。方法:采用生物信息学分析方法研究miR-362-3p的靶通路。后将143B和U2OS细胞及nu/nu雄性小鼠随机分为空白对照(BC)组,正常控制(NC)组,和过表达组(OG),CCK-8,EdU染色,伤口愈合试验,Transwell分析,采用TUNEL染色分别测定过表达miR-362-3p对细胞活力的影响,扩散,迁移,入侵,和体外143B和U2OS细胞的凋亡,肿瘤面积测定和苏木精和伊红染色分别用于确定过表达的miR-362-3p对体内OS组织生长和病理损伤的影响。qRT-PCR,蛋白质印迹,应用免疫组织化学染色分别研究体内和体外过表达miR-362-3p对OS中IL6ST/JAK2/STAT3通路的影响。结果:结合qRT-PCR的生物信息学分析方法表明IL6ST/JAK2/STAT3是miR-362-3p的靶通路之一。与NC相比,细胞活力,扩散,迁移,OG对143B和U2OS细胞的侵袭有明显的抑制作用(P<0.01),但对细胞凋亡有明显的促进作用(P<0.0001)。与NC相比,OG中OS组织的生长受到明显抑制(P<0.05),OS组织的病理损伤明显加重。IL6ST的基因表达水平,JAK2、STAT3和IL6ST的蛋白表达水平,143B和U2OS细胞中的JAK2、p-JAK2、STAT3和p-STAT3在OG中的表达低于NC(P<0.0001)。此外,IL6ST蛋白的阳性染色区域,与NC相比,OG中OS组织的JAK2,p-JAK2,STAT3和p-STAT3明显降低(P<0.01)。结论:miR362-3p的过表达通过抑制体内和体外IL6ST/JAK2/STAT3通路来减轻OS。
Objectives: To investigate the effects and the related signaling pathway of miR-362-3p on OS. Methods: The bioinformatics analysis approaches were employed to investigate the target pathway of miR-362-3p. After the 143B and U2OS cells and nu/nu male mice were randomly divided into blank control (BC) group, normal control (NC) group, and overexpression group (OG), the CCK-8, EdU staining, wound healing assay, Transwell assay, and TUNEL staining were adopted to respectively determine the effects of overexpressed miR-362-3p on the cell viability, proliferation, migration, invasion, and apoptosis of 143B and U2OS cells in vitro, tumor area assay and hematoxylin and eosin staining were employed to respectively determine the effects of overexpressed miR-362-3p on the growth and pathological injury of OS tissue in vivo. The qRT-PCR, Western blot, and immunohistochemical staining were applied to respectively investigate the effects of overexpressed miR-362-3p on the IL6ST/JAK2/STAT3 pathway in OS in vivo and in vitro. Results: The bioinformatics analysis approaches combined qRT-PCR indicated that the IL6ST/JAK2/STAT3 is one of the target pathways of miR-362-3p. Compared with NC, the cell viability, proliferation, migration, and invasion of 143B and U2OS cells were dramatically (P < 0.01) inhibited but the apoptosis was prominently (P <0 .0001) promoted in OG. Compared with NC, the growth of OS tissue was significantly (P < 0.05) suppressed and the pathological injury of OS tissue was substantially aggravated in OG. The gene expression levels of IL6ST, JAK2, and STAT3 and the protein expression levels of IL6ST, JAK2, p-JAK2, STAT3, and p-STAT3 in 143B and U2OS cells were memorably (P < 0.0001) lower in OG than those in NC. In addition, the positively stained areas of proteins of IL6ST, JAK2, p-JAK2, STAT3, and p-STAT3 of OS tissue in OG were markedly (P < 0.01) reduced compared with those in NC. Conclusion: The overexpression of miR362-3p alleviates OS by inhibiting the IL6ST/JAK2/STAT3 pathway in vivo and in vitro.