摘要
目的分析miR-363和PDZD2在骨肉瘤细胞MG-63中的功能及其相互作用,探讨miR-363对骨肉瘤细胞的调控作用及其分子机制。方法检测并对比骨肉瘤组织和瘤旁正常组织中以及成骨细胞和骨肉瘤细胞中的miR-363和PDZD2含量。检测过表达miR-363的骨肉瘤细胞MG-63的恶性生物学特性和细胞凋亡变化。利用生物信息学分析和荧光素酶报告基因检测发现miR-363的直接靶标。构建骨肉瘤动物模型,验证体外实验结果。结果骨肉瘤组织中miR-363呈现低表达,而PDZD2的表达则升高,且二者表达呈现负调控的趋势。miR-363在MG-63细胞中的过表达可以显著抑制细胞的活力,增殖和集落形成能力,促进细胞凋亡和G1/S期阻滞。生物信息学分析和荧光素酶报告基因测定表明PDZD2是miR-363的直接靶标,miR-363的过表达降低PDZD2蛋白水平,抑制PDZD2基因。动物实验研究进一步证实,miR-363具有抑制骨肉瘤生长的作用。结论miR-363在骨肉瘤的发生发展中发挥重要的调控作用,作用机制为通过负向调控PDZD2基因的表达,从而有效抑制骨肉瘤的生长。因此,miR-363可以作为治疗骨肉瘤的潜在分子靶点。
Objective To investigate the function of miR-363 and PDZD2 in osteosarcoma cell line MG-63,and the interaction between miR-363 and PDZD2 to investigate the regulation of miR-363 on osteosarcoma cells and its molecular mechanism.Methods The qPCR technique was used to detect miR-363 and PDZD2 in osteosarcoma tissues and normal tissues adjacent to the tumor.The qPCR technology was also used to detect miR-363 in osteoblasts and osteosarcoma cells,malignant biological characteristics and apoptosis of osteosarcoma cell MG-63 overexpressing miR-363 for proliferation,migration and invasion Determination of change.The nude mice were used to inhibit the osteosarcoma model in nude mice.The effect of miR-363 on the growth rate of osteosarcoma in experimental animals was observed.The expression of PDZD2 and related protein in tumor tissues were detected.Results MiR-363 in osteosarcoma tissues showed low expression,while the expression of PDZD2 increased,and the expression of both showed a negative regulation.MiR-363 was expressed in osteosarcoma cells with low expression status,and MG-63 cells were more prominent.Overexpression of miR-363 in MG-63 cells significantly inhibited cell viability,proliferation and colony forming ability,and promoted apoptosis and G1/S phase arrest.Bioinformatics analysis and luciferase reporter assays indicate that PDZD2 was a direct target of miR-363.The study found that overexpression of miR-363 reduced PDZD2 protein level,inhibited PDZD2 gene.In vivo studies further confirmed that miR-363 acts as a tumor suppressor by inhibiting tumor growth.Conclusions miR-363 plays an important regulatory role in the development of osteosarcoma.The mechanism of action is to inhibit the growth of osteosarcoma by negatively regulating the expression of PDZD2 gene.Therefore,miR-363 can be used as a potential molecular target for the treatment of osteosarcoma.
出处
《浙江临床医学》
2020年第9期1237-1240,共4页
Zhejiang Clinical Medical Journal
基金
杭州市农业与社会发展科研自主申报项目(20191203B88)。
