TFESB调控lncRNA LINC00858/miR-324-5p通路抑制宫颈癌细胞的增殖、迁移和侵袭

Total flavonoids extract from Sculellaria barbata D.Don inhibits the proliferation,migration and invasion of cervical cancer cells by regulating lncRNA LINC00858/miR-324-5p pathway

目的:探讨半枝莲总黄酮提取物(total flavonoids extract from Scutellaria barbata D.Don,TFESB)调控长链非编码RNA(lncRNA)LINC00858/miR-324-5p通路对宫颈癌细胞增殖、迁移和侵袭的影响及其机制。方法:qRT-PCR检测20例宫颈癌组织和与其对应的癌旁组织中LINC00858和miR-324-5p的表达水平。以宫颈癌SiHa细胞为研究对象,将SiHa细胞随机分为Con组、TFESB处理组、si-NC组、si-LINC00858组、miR-NC组、miR-324-5p组、TFESB+pcDNA组和TFESB+pcDNA-LINC00858组。qRT-PCR检测各组细胞中LINC00858和miR-324-5p的表达水平。细胞计数(CCK-8)试剂盒检测细胞增殖活力。Transwell实验检测细胞的迁移和侵袭能力。Western blot检测增殖相关蛋白(CyclinD1、p21)和迁移侵袭相关蛋白(MMP-2、MMP-9)的表达水平。双荧光素酶报告基因实验和qRT-PCR验证LINC00858和miR-324-5p的靶向调控关系。结果:与癌旁组织相比,宫颈癌组织中LINC00858的表达水平显著升高,miR-324-5p的表达水平显著降低。与Con组比较,TFESB处理组SiHa细胞LINC00858的表达水平显著降低,miR-324-5p的表达水平显著升高,细胞的增殖、迁移和侵袭能力显著降低,并呈显著的浓度依赖性;与si-NC组比较,si-LINC00858组SiHa细胞的增殖、迁移和侵袭能力显著降低;与miR-NC组比较,miR-324-5p组SiHa细胞的增殖、迁移和侵袭能力显著降低;与TFESB+pcDNA组比较,TFESB+pcDNA-LINC00858组SiHa细胞的增殖、迁移和侵袭能力显著提高。结论:半枝莲总黄酮提取物可抑制宫颈癌细胞的增殖、迁移和侵袭能力,其机制与调控lncRNA LINC00858/miR-324-5p通路有关。

Objective:To investigate the effect of total flavonoids extract from Sculellaria barbata D.Don(TFESB)on the proliferation,migration and invasion of cervical cancer cells by regulating lncRNA LINC00858/miR-324-5p pathway.Methods:qRT-PCR was used to detect the expression levels of LINC00858 and miR-324-5p in 20 cervical cancer tissues and their corresponding adjacent tissues.SiHa cells were randomly divided into Con group,TFESB treatment group,si-NC group,si-LINC00858 group,miR-NC group,miR-324-5p group,TFESB+pcDNA group and TFESB+LINC00858 group.The expression levels of LINC00858 and miR-324-5p in each group of SiHa cells were detected by qRT-PCR.The cell proliferation activity was measured by the CCK-8 method.Transwell assay was used to detect cell migration and invasion ability.The expression levels of proliferation-associated proteins(CyclinD1,p21)and migration-invasive related proteins(MMP-2,MMP-9)were detected by Western blot.The dual luciferase reporter assay and qRT-PCR were to verify the targeted and regulatory relationship between LINC00858 and miR-324-5p.Results:Compared with adjacent tissues,the expression level of LINC00858 in cervical cancer tissues was significantly increased,while the expression level of miR-324-5p was significantly decreased.Compared with the Con group,the expression level of LINC00858 in SiHa cells treated with TFESB was significantly decreased,the expression level of miR-324-5p was significantly increased,the proliferation,migration and invasion ability of the cells were significantly decreased,and showed a significant concentration-dependent.Compared with the si-NC group,the proliferation,migration and invasion ability of SiHa cells in the si-LINC00858 group were significantly decreased.Compared with the miR-NC group,the proliferation,migration and invasion ability of SiHa cells in the miR-324-5p group were significantly reduced.Compared with the TFESB+pcDNA group,the proliferation,migration and invasion ability of SiHa cells in the TFESB+pcDNA-LINC00858 group were significantly increased.Conclusion:The total flavonoid extract from Sculellaria barbata D.Don.inhibits the proliferation,migration and invasion of cervical cancer cells,and its mechanism is related to the regulation of lncRNA LINC00858/miR-324-5p pathway.