氧化槐果碱通过调控lncRNA DUXAP8/miR-519b-3p轴影响宫颈癌细胞增殖、凋亡、迁移及侵袭的实验研究  

Experimental Study of Oxysophocarpine on Proliferation,Apoptosis, Migration and Invasion of Cervical Cancer Cells by Regulating LncRNA DUXAP8/miR-519b-3p Axis

作　　者：姜烨 杨茹 陈杰 赵婷婷 石伟 罗莉 李功卓 JIANG Ye;YANG Ru;CHEN Jie;ZHAO Tingting;SHI Wei;LUO Li;LI Gongzhuo(Department of Oncology,Guiyang Hospital of Guihang,Guiyang 550000,China)

机构地区：[1]贵航贵阳医院,肿瘤科,贵阳550000

出　　处：《中国细胞生物学学报》2021年第4期736-746,共11页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81660749);贵阳市科技计划项目(批准号:[2019]2-18)资助的课题。

摘　　要：为了探讨氧化槐果碱(OSC)对宫颈癌HeLa细胞增殖、凋亡、迁移和侵袭的影响及作用机制,本研究将HeLa细胞分为对照组(Con组)、OSC组、转染DUXAP8小干扰RNA(si-DUXAP8)组、转染小干扰RNA阴性对照(si-NC)组、OSC+DUXAP8过表达载体(pcDNA-DUXAP8)组和OSC+空载体(pcDNA)组,然后用MTT法检测细胞增殖,Transwell检测细胞迁移和侵袭,流式细胞术检测细胞凋亡,蛋白印迹法检测细胞中CyclinD1、p21、Bcl-2、Bax、MMP-2和MMP-9蛋白水平,RT-qPCR检测细胞中DUXAP8和miR-519b-3p水平;双荧光素酶报告基因实验验证DUXAP8和miR-519b-3p调控关系。结果显示,与Con组比较,OSC组HeLa细胞抑制率、凋亡率、Bax和p21蛋白水平、miR-519b-3p水平升高(P<0.05),细胞迁移数、侵袭数、CyclinD1、Bcl-2、MMP-2和MMP-9的蛋白水平和DUXAP8水平降低(P<0.05),且呈剂量依赖性(P<0.05);与si-NC组比较,si-DUXAP8组HeLa细胞抑制率、凋亡率和Bax、p21蛋白水平升高(P<0.05),细胞迁移和侵袭数及CyclinD1、Bcl-2、MMP-2、MMP-9的蛋白水平降低(P<0.05);DUXAP8在HeLa细胞中靶向负调控miR-519b-3p表达;与OSC+pcDNA组比较,OSC+pcDNA-DUXAP8组HeLa细胞抑制率、凋亡率和Bax、p21的蛋白水平降低(P<0.05),细胞迁移和侵袭数及CyclinD1、Bcl-2、MMP-2、MMP-9的蛋白水平升高(P<0.05)。这提示氧化槐果可能通过调DUXAP8/miR-519b-3p轴抑制宫颈癌HeLa细胞增殖、迁移和侵袭,并诱导细胞凋亡。In order to explore the effects of OSC (Oxysophocarpine) on proliferation,apoptosis,migration and invasion of cervical cancer HeLa cells,this study divided HeLa cells into Con group (control group),OSC group,si-DUXAP8 (DUXAP8 small interfering RNA) group,si-NC (small interfering RNA negative control) group,OSC+pcDNA-DUXAP8 (DUXAP8 overexpression vector) group and OSC+pcDNA group.Then cell proliferation was detected by MTT assay;cell migration and invasion were detected by Transwell;cell apoptosis was detected by flow cytometry;the protein levels of CyclinD1,p21,Bcl-2,Bax,MMP-2 and MMP-9 were detected by Western blot;the levels of DUXAP8 and miR-519b-3p were detected by RT-qPCR.The regulatory relationship between DUXAP8 and miR-519b-3p was verified by dual luciferase reporter gene assay.The results showed that compared with the Con group,the inhibition rate,apoptosis rate,the protein levels of Bax and p21,and the level of miR-519b-3p in the OSC groups were increased (P<0.05),while the number of cell migration and invasion,the protein levels of CyclinD1,Bcl-2,MMP-2 and MMP-9,and the level of DUXAP8 were decreased (P<0.05).Compared with the si-NC group,the inhibition rate,apoptosis rate,and the protein levels of Bax and p21 in the si-DUXAP8 group were increased (P<0.05),while the number of cell migration and invasion,and the protein levels of CyclinD1,Bcl-2,MMP-2 and MMP-9 were decreased (P<0.05).DUXAP8 negatively regulated the expression of miR-519b-3p in HeLa cells.Compared with the OSC+pcDNA group,the inhibition rate,apoptosis rate,and the protein levels of Bax and p21 in the OSC+pcDNA-DUXAP8 group were decreased (P<0.05),while the number of cell migration and invasion,and the protein levels of CyclinD1,Bcl-2,MMP-2 and MMP-9 were increased (P<0.05).This suggested that OSC could inhibit the proliferation,migration and invasion of cervical cancer HeLa cells and induce their apoptosis by regulating DUXAP8/miR-519b-3p axis.

关 键 词：氧化槐果碱 lncRNA DUXAP8 miR-519b-3p 宫颈癌 细胞增殖 迁移侵袭 凋亡 

分 类 号：R285[医药卫生—中药学]