lncRNA MIR34AHG通过miR-296-5p/SRCIN1轴影响喉癌细胞的恶性生物学行为

LncRNA MIR34AHG Affects the Malignant Biological Behavior of Laryngeal Cancer Cells through the Mir-296-5p/SRCIN1 Axis

目的探讨长链非编码RNA(lncRNA)MIR34AHG通过miR-296-5p/SRCIN1轴对喉癌细胞增殖和迁移的作用。方法qPCR检测MIR34AHG在喉癌细胞系(TU686、Hep-2、TU177、AMC-NH-8、TU212)以及支气管上皮细胞系(16HBE)中的表达。选取MIR34AHG表达最低细胞系,分别构建MIR34AHG过表达细胞系(MIR34AHG组)和对照细胞系(对照组)。MTT法、细胞划痕实验分别检测上调MIR34AHG对细胞增殖和迁移的影响。生物信息学工具预测MIR34AHG的靶基因,双荧光素酶报告基因实验检测MIR34AHG与靶基因之间的相互作用。qPCR和Western blot法检测上调MIR34AHG对相关基因和蛋白表达的影响。结果与16HBE细胞比较,喉癌细胞系中MIR34AHG呈低表达(P<0.01),且以TU686细胞中表达最低(P<0.01)。与对照组比较,MIR34AHG过表达抑制TU686细胞的增殖和迁移(P<0.05)。MIR34AHG和miR-296-5p之间存在靶向关系(P<0.01),miR-296-5p的靶基因可能是SRCIN1。与对照组比较,MIR34AHG过表达抑制miR-296-5p表达(P<0.01),促进SRCIN1基因的表达(P<0.01)。结论MIR34AHG可能通过下调miR-296-5p的表达、上调SRCIN1的表达从而抑制喉癌细胞的增殖和迁移。

Objective To explore the effect of long non-coding RNA(lncRNA)MIR34AHG on the proliferation and migration of laryngeal cancer cells through the miR-296-5p/SRCIN1 axis.Methods qPCR was used to detect the expression of MIR34AHG in laryngeal cancer cell lines(TU686,Hep-2,TU177,AMC-NH-8,TU212)and bronchial epithelial cell lines(16HBE).The cell line with the lowest MIR34AHG expression was selected,and the MIR34AHG over-expressing cell line(MIR34AHG group)and control cell line(control group)were constructed respectively.MTT method and cell scratch test were used to detect the effect of up-regulation of MIR34AHG on cell proliferation and migration.Bioinformatics tools were used to predict the target gene of MIR34AHG,and the dual luciferase reporter gene experiment was used to detect the interaction between MIR34AHG and target gene.qPCR and Western blot were used to detect the effect of up-regulation of MIR34AHG on the expression of related genes and proteins.Results Compared with 16HBE cells,the expression of MIR34AHG in the laryngeal carcinoma cell lines was low(all P<0.01),and the expression in TU686 cells was the lowest(P<0.01).Compared with the control group,MIR34AHG overexpression inhibited the proliferation and migration of TU686 cells(all P<0.05).There is a targeting relationship between MIR34AHG and miR-296-5p(P<0.01),and the target gene of miR-296-5p may be SRCIN1.Compared with the control group,overexpression of MIR34AHG inhibited the expression of miR-296-5p(P<0.01)and promoted the expression of SRCIN1 gene(P<0.01).Conclusion MIR34AHG may inhibit the proliferation and migration of laryngeal cancer cells by down-regulating the expression of miR-296-5p and up-regulating the expression of SRCIN1.