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目的研究miR-27b在胶质瘤细胞系U87中的表达、功能及作用机制。方法在多个胶质瘤细胞系及胶质瘤标本中检测miR-27b的表达;利用反义miR-27b转染U87,下调miR-27b的表达,应用流式细胞术和MTT法,评价细胞生长、增殖、凋亡及周期变化;采用荧光素酶法及Western印迹法检测miR-27b作用的初步机制及通路蛋白的变化。结果 miR-27b在胶质瘤细胞系及胶质瘤标本中均呈高表达;转染反义miR-27b后,表达降低86%,MTT法显示转染反义miR-27b后细胞生长受到抑制且凋亡细胞增加13%;荧光素酶实验结果显示:miR-27b发挥作用可能与β-catenin/Tcf-4通路相关,Western印迹法显示通路相关蛋白STAT3、c-myc和Cyclin D1在转染反义miR-27b后表达下降。结论 miR-27b可促进人脑胶质瘤细胞生长、增殖,具有原癌基因的作用,其发挥作用可能与β-catenin/Tcf-4通路密切相关。
Objective To study the expression, function and mechanism of miR-27b in glioma cell line U87. Methods The expression of miR-27b was detected in multiple glioma cell lines and glioma specimens. The expression of miR-27b was down-regulated by using U87 antisense miR-27b. Flow cytometry and MTT were used to evaluate the expression of miR- Growth, proliferation, apoptosis and cycle changes. The primary mechanism of miR-27b and the changes of the pathway proteins were detected by luciferase and Western blot. Results miR-27b was highly expressed in both glioma cell lines and glioma specimens. After transfection with antisense miR-27b, the expression of miR-27b was decreased by 86%. MTT assay showed that the growth of cells was inhibited after transfection with antisense miR-27b The apoptotic cells increased by 13%. The results of luciferase assay showed that miR-27b may play a role in β-catenin / Tcf-4 pathway. Western blotting showed that STAT3, c-myc and Cyclin D1 Antisense miR-27b expression decreased. Conclusion miR-27b can promote the growth and proliferation of human glioma cells and has the function of protooncogene. Its function may be closely related to the β-catenin / Tcf-4 pathway.