苦参碱通过lncRNA CASC11/miR-381-3p轴调控宫颈癌细胞的恶性生物学行为的研究  

Matrine regulates the malignant biological behavior of cervical cancer cells through lncRNA CASC11/ miR-381-3p axis

作　　者：孙海燕 刘文博[1] 王立娟 孙亚男 刘东芳 SUN Haiyan;LIU Wenbo;WANG Lijuan;SUN Yanan;LIU Dongfang(Department of Gynecology,Chengde Central Hospital,Chengde,067000,China)

机构地区：[1]承德市中心医院妇科,河北承德067000

出　　处：《沈阳药科大学学报》2023年第2期192-198,共7页Journal of Shenyang Pharmaceutical University

基　　金：河北省承德市科学技术研究与发展课题计划资助项目(202002A079)

摘　　要：目的探讨苦参碱是否通过调控lncRNA CASC11/miR-381-3p的表达,调控宫颈癌细胞的增殖、凋亡、迁移和侵袭。方法宫颈癌细胞SiHa分别转染pcDNA3.1空载体、pcDNA3.1-lncRNA CASC11、mimics阴性对照和miR-381-3p mimics,经不同质量浓度(25、50和100 mg·L^(-1))的苦参碱处理后,采用qRT-PCR试验检测细胞中lncRNA CASC11和miR-381-3p表达,CCK-8试验检测细胞存活率,流式细胞仪检测细胞凋亡率,Transwell小室试验检测细胞迁移和侵袭,双荧光素酶试验验证lncRNA CASC11和miR-381-3p的靶向关系,Western blot试验检测增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、Cleaved-caspase3、E-cadherin和N-cadherin蛋白表达。结果lncRNA CASC11在宫颈癌细胞中高表达,miR-381-3p在宫颈癌细胞中低表达;不同质量浓度的苦参碱均能抑制SiHa细胞增殖、迁移和侵袭,并促进细胞凋亡,且具有浓度依赖性;不同质量浓度的苦参碱均能抑制SiHa细胞中lncRNA CASC11表达,促进细胞中miR-381-3p的表达,且具有浓度依赖性;SiHa细胞中lncRNA CASC11靶向负调控miR-381-3p的表达;过表达lncRNA CASC11能够逆转苦参碱对SiHa细胞增殖、凋亡、迁移和侵袭的作用,而同时过表达miR-381-3p则能逆转lncRNA CASC11对苦参碱的回复作用。结论苦参碱通过抑制lncRNA CASC11的表达,进而上调miR-381-3p的表达,发挥对宫颈癌细胞增殖、迁移和侵袭的抑制作用,并促进细胞凋亡。Objective To investigate whether matrine regulates the proliferation,apoptosis,migration and invasion of cervical cancer cells by regulating the expression of lncRNA CASC11/miR-381-3p.Methods Cervical cancer cells SiHa were transfected with pcDNA3.1 empty vector,PCDNA3.1-lncrNA CASC11,negative control of mimics,and miR-381-3p mimics.After matrine treatment at different concentrations(25 mg·L^(-1),50 mg·L^(-1),and 100 mg·L^(-1)),the expression of lncRNA CASC11 and miR-381-3p in the cells was detected by QRT-PCR.Cell survival rate was detected by CCK8 experiment,and cell apoptosis rate was detected by flow cytometry.Cell migration and invasion were detected by Transwell assay,the targeted relationship between lncRNA CASC11 and miR-381-3p was verified by double luciferase assay,and protein expressions of PCNA,Cleaved caspase-3,E-cadherin and N-cadherin were detected by Western blot assay.Results LncRNA CASC11 was highly expressed in cervical cancer cells,while miR-381-3p was low in cervical cancer cells.Matrine at different concentrations can inhibit the proliferation,migration and invasion of SiHa cells and promote apoptosis in a concentration-dependent manner.Matrine at different concentrations can inhibit the expression of lncRNA CASC11 in SiHa cells and promote the expression of miR-381-3p in cells in a concentration-dependent manner.LncRNA CASC11 negatively regulates the expression of miR-381-3p in SiHa cells.Overexpression of lncRNA CASC11 can reverse the effect of matrine on proliferation,apoptosis,migration and invasion of SiHa cells,while simultaneous overexpression of miR-381-3p can reverse the recovery effect of lncRNA CASC11 on matrine.Conclusions Matrine could inhibit the proliferation,migration and invasion of cervical cancer cells and promote apoptosis by inhibiting the expression of lncRNA CASC11 and then up-regulating the expression of miR-381-3p.

关 键 词：宫颈癌 苦参碱 长链非编码RNA CASC11 miR-381-3p 

分 类 号：R739.4[医药卫生—肿瘤]