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目的:探讨下调miR-27a对U87胶质瘤细胞的影响。方法 :分别用慢病毒干扰质粒miRZip-27a anti-miR-27a mi-croRNA construct或慢病毒空质粒pGreenPuro Scramble Hairpin Control-Construct与慢病毒包装质粒混合物共转染293T细胞,获得慢病毒颗粒命名为Lt-I和Lt-NC。然后分别用慢病毒Lt-I和Lt-NC感染U87胶质瘤细胞,通过流式细胞仪筛选出稳定感染的U87胶质瘤细胞。通过定量PCR法分别检测稳定感染的U87细胞和未感染的空白U87细胞miR-27a,CCK-8法检测胶质瘤细胞增殖情况,Transwell侵袭试验检测胶质瘤细胞的侵袭能力。结果:相对于未感染慢病毒的空白U87胶质瘤细胞,稳定感染Lt-I的U87胶质瘤细胞miR-27a表达明显下调、细胞增殖减缓、侵袭能力降低,而感染Lt-NC组(阴性对照组)则未见此改变。结论:miR-27a的下调能有效抑制U87胶质瘤细胞的增殖,降低U87胶质瘤细胞的侵袭能力。
Objective: To investigate the effect of down-regulating miR-27a on U87 glioma cells. METHODS: 293T cells were co-transfected with the lentiviral plasmid miRZip-27a anti-miR-27a mi-croRNA construct or the lentiviral plasmid pGreenPuro Scramble Hairpin Control-Construct and the lentivirus packaging plasmid mixture respectively. The lentiviral particles were named as Lt -I and Lt-NC. U87 glioma cells were then infected with lentivirus Lt-I and Lt-NC, and stable infected U87 glioma cells were screened by flow cytometry. The stable infected U87 cells and uninfected blank U87 cells miR-27a were detected by quantitative PCR. The proliferation of glioma cells was detected by CCK-8 assay. The invasion ability of glioma cells was detected by Transwell invasion assay. Results: The expression of miR-27a in U87 glioma cells stably infected with Lt-I was significantly lower than that in untreated lentivirus-infected U87 glioma cells, the cell proliferation was slowed and the invasion ability was decreased. However, the infection of Lt-NC group (negative Control group) did not see this change. Conclusion: Down-regulation of miR-27a can effectively inhibit the proliferation of U87 glioma cells and reduce the invasion ability of U87 glioma cells.