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作 者:乐曲 薛一雪[2] 阮雪蕾 刘云会[1] YUE Qu;XUE Yi-xue;RUAN Xue-lei;LIU Yun-hui(Department of Neurosurgery,Shengjing Hosipital,Shenyang 110004;Department of Neurobiology of School of Life Sciences,China Medical University,Shenyang 110122,China)
机构地区:[1]中国医科大学附属盛京医院神经外科,辽宁沈阳110004 [2]中国医科大学生命科学学院神经生物学教研室,辽宁沈阳110122
出 处:《解剖科学进展》2022年第3期244-248,共5页Progress of Anatomical Sciences
基 金:国家自然科学基金(81672511,81872073);辽宁省科学技术计划项目(2017225020);盛京自由研究者计划项目(201802)。
摘 要:目的 探讨miR-202-5p调控胶质瘤细胞恶性生物学行为的潜在分子机制。方法 应用实时定量PCR实验检测miR-202-5p在人脑胶质瘤U87MG细胞中的内源性表达;实时定量PCR实验和Western blot实验用于检测TSPAN12在人脑胶质瘤U87MG细胞的内源性表达;应用CCK-8、transwell、流式细胞术检测过表达miR-202-5p或沉默TSPAN12对U87MG细胞增殖、迁移、侵袭和凋亡能力的影响;应用双荧光素酶报告基因分析系统检测miR-202-5p和TSPAN12的结合作用。结果 miR-202-5p在U87MG细胞中低表达,TSPAN12在U87MG细胞中高表达。过表达miR-202-5p或沉默TSPAN12显著抑制U87MG细胞的增殖、迁移、侵袭,促进细胞凋亡。miR-202-5p靶向结合TSPAN12 mRNA的3’UTR。结论 miR-202-5p通过靶向结合TSPAN12抑制胶质瘤细胞的恶性生物学行为。Objective To investigate the potential molecular mechanism that miR-202-5p regulated the malignant biological behavior of glioma U87MG cells. Methods Real-time PCR was applied to analyze the miR-202-5p expression in U87MG cells. Real-time PCR and Western blot were applied to analyze the TSPAN12 expression in U87MG cells. CCK-8, transwell and flow cytometry assays were applied to evaluate the effects of miR-202-5p overexpression or TSPAN12 knockdown on proliferation, migration, invasion and apoptosis ability of U87MG cells. Luciferase reporter assay was applied to verify the binding site between miR-202-5p and TSPAN12. Results MiR-202-5p was downregulated in U87MG cells compared with normal human astrocytes(NHA), while TSPAN12 was upregulated in U87MG cells compared with NHA. MiR-202-5p overexpression or TSPAN12 knockdown inhibited the malignant behavior of U87MG cells. MiR-202-5p directly targeted the 3’UTR of TSPAN12 mRNA. Conclusion MiR-202-5p inhibited the malignant biological behavior of glioma cells via targeting TSPAN12.
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