LncRNA FAM83H-AS1调控miR-136-5p/HOXC10轴对胶质瘤细胞增殖和凋亡的影响

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目的探讨长链非编码RNA(LncRNA)序列相似性家族83成员H反义RNA 1(lncRNA FAM83H-AS1)调控微小RNA-136-5p（miR-136-5p）/同源异型盒基因10(HOX10)轴对胶质瘤细胞增殖和凋亡的影响。方法体外培养人胶质瘤细胞U87，对U87细胞转染质粒，并将细胞分为对照组、si-NC组、si-FAM83H-AS1组、miR-NC组、miR-136-5p mimics组、si-FAM83H-AS+NC inhibitor组、si-FAM83H-AS+miR-136-5p inhibitor组、miR-136-5p mimics+HOXC10 NC组、miR-136-5p mimics+HOXC10组。检测胶质瘤组织与细胞U87中FAM83H-AS1、miR-136-5p、HOXC10的表达。细胞计数盒8（CCK-8）法检测U87细胞增殖；流式细胞仪检测U87细胞凋亡；蛋白免疫印迹（WB）法检测U87细胞中HOXC10、Cyclin D1、Bcl-2、Bax蛋白的表达；双荧光素酶报告系统分析miR-136-5p与FAM83H-AS1、HOXC10的靶向关系。结果胶质瘤组织中FAM83H-AS1表达水平显著升高，miR-136-5p表达水平降低（P<0.05）；沉默FAM83H-AS1表达或过表达miR-136-5p，U87细胞凋亡率、Bax蛋白表达显著升高，细胞存活率、HOXC10 mRNA、Cyclin D1、Bcl-2蛋白表达显著降低（P<0.05）；双荧光素酶报告基因显示，miR-136-5p与FAM83H-AS1、HOXC10均有靶向关系；抑制miR-136-5p表达可逆转沉默FAM83H-AS1表达对U87细胞增殖的抑制作用，HOXC10过表达可逆转过表达miR-136-5p对U87细胞增殖的抑制作用。结论FAM83H-AS1通过调控miR-136-5p/HOXC10轴促进胶质瘤细胞增殖，抑制细胞凋亡。

Objective To investigate the effects of long non-coding RNA(LncRNA) family with sequence similarity 83, member H antisense RNA1(lncRNA FAM83H-AS1) on the proliferation and apoptosis of glioma cells by regulating microRNA-136-5p(miR-136-5p)/homeobox C10(HOX10) axis. Methods Human glioma cells U87 were cultured in vitro and transfected with plasmids, then the cells were divided into control group, si-NC group, si-FAM83H-AS1 group, miR-NC group, miR-136-5p mimics group, si-FAM83H-AS+NC inhibitor group, si-FAM83H-AS+miR-136-5p inhibitor group, miR-136-5p mimics+HOXC10 NC group, miR-136-5p mimics+HOXC10 group. The expression of FAM83H-AS1, miR-136-5p, and HOXC10 in glioma tissues and U87 cells were detected. Cell counting box 8(CCK-8) method was used to detect U87 cell proliferation; flow cytometry was used to detect U87 cell apoptosis. Western Blot(WB) method was used to detect the expression of HOXC10, Cyclin D1, Bcl-2, and Bax protein in U87 cells. The dual luciferase reporter system was used to analyze the targeting relationship between miR-136-5p and FAM83H-AS1 and HOXC10. Results The expression level of FAM83H-AS1 in glioma tissue was significantly increased, and the expression level of miR-136-5p was decreased(P<0.05). After silencing FAM83H-AS1 expression or overexpression of miR-136-5p, the U87 cell apoptosis rate and Bax protein expression increased significantly, the cell survival rate, HOXC10 mRNA, Cyclin D1, Bcl-2 protein expression decreased significantly(P<0.05). The dual luciferase reporter gene showed that miR-136-5p had a targeting relationship with FAM83H-AS1 and HOXC10. Inhibition of miR-136-5p expression reversed the inhibitory effect of silencing FAM83H-AS1 expression on U87 cell proliferation. Overexpression of HOXC10 could reverse the inhibitory effect of overexpression of miR-136-5p on U87 cell proliferation. Conclusion FAM83H-AS1 promotes the proliferation of glioma cells and inhibits apoptosis by regulating the miR-136-5p/HOXC10 axis.