微小RNA-301b-3p靶向第10号染色体缺失的磷酸酶及张力蛋白同源物对骨肉瘤细胞增殖、凋亡的影响  

Effect of miR-301b-3p targeting PTEN on proliferation and apoptosis of osteosarcoma cells

作　　者：卿海辉 张小舟 胡敏[1] 贺茂林[2] QING Haihui;ZHANG Xiaozhou;HU Min;HE Maolin(Department of Orthopedics Ward Three,Xiaogan Hospital,Wuhan University of Science and Technology,Xiaogan,Hubei 432000,China;Department of Spine and Bone Diseases,The First Affiliated Hospital of Guangxi Medical University,Nanning,Guangxi Zhuang Autonomous Region 530021,China)

机构地区：[1]武汉科技大学附属孝感医院骨科三病区,湖北孝感432000 [2]广西医科大学第一附属医院脊柱骨病外科,广西壮族自治区南宁530021

出　　处：《安徽医药》2021年第10期1957-1961,共5页Anhui Medical and Pharmaceutical Journal

基　　金：广西科技厅科学研究与技术开发资助项目(桂科攻1140003A-14)。

摘　　要：目的探讨微小RNA-301b-3p(miR-301b-3p)对骨肉瘤细胞增殖、凋亡的影响及其作用机制。方法本研究起止时间为2019年3—9月。人成骨细胞(hFOB)和骨肉瘤细胞U2OS、MG63购自美国菌种保藏中心。U2OS细胞分为miRNA抑制物阴性对照(anti-miR-NC)组、miR-301b-3p抑制物(anti-miR-301b-3p)组、anti-miR-301b-3p+小干扰RNA阴性对照(si-NC)组、anti-miR-301b-3p+第10号染色体缺失的磷酸酶及张力蛋白同源物(PTEN)小干扰RNA(si-PTEN)组;实时荧光定量PCR(RT-qP-CR)检测miR-301b-3p表达水平;四甲基偶氮唑盐比色法(MTT)检测细胞活性;流式细胞术检测细胞凋亡;双荧光素酶报告实验检测miR-301b-3p和PTEN的靶向关系。结果与h FOB细胞相比,U2OS、MG63中miR-301b-3p表达水平[(0.89±0.09),(0.70±0.07)比(0.20±0.02)]显著升高。与anti-miR-NC组比较,anti-miR-301b-3p组U2OS细胞活性[(0.59±0.06)比(1.33±0.13)]显著降低,而凋亡率[(22.06±2.31)%比(7.48±0.78)%]、PTEN蛋白表达[(0.69±0.07)比(0.35±0.03)]显著升高。PTEN是miR-301b-3p的直接靶基因。与anti-miR-301b-3p+si-NC组比较,anti-miR-301b-3p+si-PTEN组U2OS细胞活性[(1.16±0.12)比(0.56±0.06)]显著升高,而凋亡率[(10.28±1.16)比(22.12±2.34)]显著降低。结论抑制miR-301b-3p表达可通过调控PTEN抑制骨肉瘤细胞增殖,促进细胞凋亡。Objective To explore the effect of microRNA-301b-3p(miR-301b-3p)on proliferation and apoptosis of osteosarcoma cells and its mechanism.Methods The start and end time of this research was from March to September 2019.Human osteoblasts(hFOB)and osteosarcoma cells U20 S and MG63 were purchased from the American Tissue Culture Collection.U20 S cells were assigned into miRNA inhibitor negative control(anti-miR-NC)group,miR-301b-3p inhibitor(anti-miR-301b-3p)group,anti-miR-301b-3p+small interfering RNA negative control(si-NC)group,anti-miR-301b-3p+phosphatase and tensin homolog deleted on chromosome10(PTEN)small interfering RNA(si-PTEN)group.Real-time quantitative PCR(RT-qPCR)was used to detect the expression of miR-301b-3p,methyl thiazolyl tetrazolium(MTT)assay was used to detect cell viability,flow cytometry was used to detect apoptosis,and dual luciferase reporter assay was used to detect the targeting relationship between miR-301b-3p and PTEN.Results Compared with hFOB cells,the expressions of miR-301b-3p[(0.89±0.09),(0.70±0.07)vs(0.20±0.02)]in U20 S and MG63 cells were significantly increased.Compared with the anti-miR-NC group,the cell viability[(0.59±0.06)vs(1.33±0.13)]of U20 S cells in the anti-miR-301b-3p group was significantly reduced,while the apoptosis rate[(22.06±2.31)%vs(7.48±0.78)%]and PTEN protein expression[(0.69±0.07)vs(0.35±0.03)]were significantly increased.PTEN is the direct target gene of miR-301b-3p.Compared with the anti-miR-301b-3p+si-NC group,the cell viability[(1.16±0.12)vs(0.56±0.06)]of U20 S cells in the anti-miR-301b-3p+si-PTEN group was significantly increased,while the apoptosis rate[(10.28±1.16)%vs(22.12±2.34)%]was significantly reduced.Conclusion Inhibition of miR-301b-3p expression can inhibit osteosarcoma cell proliferation and promote cell apoptosis by regulating PTEN.

关 键 词：骨肉瘤 微小RNA-301b-3p 第10号染色体缺失的磷酸酶及张力蛋白同源物(PTEN) 细胞增殖 细胞凋亡 

分 类 号：R738.1[医药卫生—肿瘤]