孙海燕,刘文博,王立娟,孙亚男,刘东芳
SUN Haiyan,LIU Wenbo,WANG Lijuan,SUN Yanan,LIU Dongfang

• 1:承德市中心医院妇科

摘要(Abstract)：

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(KeyWords)： 宫颈癌;苦参碱;长链非编码RNA CASC11;miR-381-3p
cervical cancer;matrine;long non-coding RNA;miR-381-3p

Abstract:

Keywords:

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

作者(Author): 孙海燕,刘文博,王立娟,孙亚男,刘东芳
SUN Haiyan,LIU Wenbo,WANG Lijuan,SUN Yanan,LIU Dongfang

DOI: 10.14066/j.cnki.cn21-1349/r.2020.1317

参考文献(References)：

• [1] WANG Y,SONG S F,LIU F.Research progress of epidemiological characteristics and risk factors of cervical cancer in China[J].Maternal & Child Health Care of China(中国妇幼保健),2019,34(5):1206-1208.
• [2] HILLEMANNS P,SOERGEL P,HERTEL H,et al.Epidemiology and early detection of cervical cancer[J].Oncol Res Treat,2016,39(9):501-516.
• [3] WU W G.Localization and development of TCM in the era of tumor targeting drug therapy[J].Chinese Journal of Integrated Traditional and Western Medicine(中国中西医结合杂志),2018,38(8):12-14.
• [4] WU X L,ZHOU J,CAI D G,et al.Matrine inhibits the metastatic properties of human cervical cancer cells via downregulating the p38 signaling pathway[J].Oncol Rep,2017,38(2):1312-1320.
• [5] YAN W M,GAO Y,YU Z L,et al.Effect of matrine on the proliferation of SiHa cells and the negative regulation of JAK-SATA pathway[J].The Chinese Journal of Clinical Pharmacology(中国临床药理学杂志),2018,34(10):53-56.
• [6] CHEN G T,YANG X,CHEN Y X.Different Expression profiles of lncRNA in cervical cancer tissues and its significance[J].The Practical Journal of Cancer(实用癌症杂志),2017,32(4):523-527.
• [7] DU M Y,YI L,HE X.Research progress and clinical prospect of LncRNA in tumor invasion and metastasis[J].Journal of Chinese Oncology(肿瘤学杂志),2017,23(1):54-59.
• [8] HSU W H,LIU L F,CHEN X,et al.LncRNA CASC11 promotes the cervical cancer progression by activating Wnt/beta-catenin signaling pathway[J].Biol Res,2019,52(1):33-39.
• [9] ZHANG M F,SHEN Y Q.Research advances on pharmacological actions of matrine against cervical carcinoma and endometrial carcinoma[J].Anti-infection Pharmacy(抗感染药学),2019,16(6):925-930.
• [10] ZHAO H,GUO J H,CUI Y,et al.Research progress on structural modifications of matrine and its anticancer activity[J].Drugs & Clinic(现代药物与临床),2015,30(5):600-604.
• [11] ZHU Y,SONG X.The advance of the interaction between miRNA and lncRNA in cancer reseach[J].Basic & Clinical Medicine(基础医学与临床),2015,35(11):114-118.
• [12] LUAN X,WANG Y.LncRNA XLOC_006390 facilitates cervical cancer tumorigenesis and metastasis as a ceRNA against miR-331-3p and miR-338-3p[J].J Gynecol Oncol,2018,29(6):e95-e101.
• [13] LI Z,WENQUAN K,XIAOLAN L,et al.LncRNA CASC11 promoted gastric cancer cell proliferation,migration and invasion in vitro by regulating cell cycle pathway[J].Cell Cycle (Georgetown,Tex.),2018,17(15):1-15.
• [14] HUARONG L,CHENG D X,WEI L,et al.lncRNA CASC11 promotes cancer cell proliferation in bladder cancer through miRNA-150[J].J Cell Biochem,2019,120(8):13487-13493.
• [15] NAN C,JU W,MIN Y,et al.LncRNA CASC11 promotes cancer cell proliferation in hepatocellular carcinoma by inhibiting miRNA-188-5p[J].Bioscience Rep,2019,39(4):251-262.
• [16] WANG M Y,WANG G L,CHEN X L,et al.Progress on biological functions of miRNA and its diagnosis and treatment in tumor[J].Progress in Veterinary Medicine(动物医学进展),2018,39(1):95-98.
• [17] YU S L,YU Y J,WANG G Y,et al.Research progress on the correlation between miRNA and disease[J].Journal of Mudanjiang Medical University(牡丹江医学院学报),2015,36(1):94-96.
• [18] SHANG A Q,ZHOU C,BIAN G X,et al.miR-381-3p restrains cervical cancer progression by downregulating FGF7[J].J Cell Biochem,2019,120(1):778-789.
• [19] KONG W,YANG L,LI P P,et al.miR-381-3p inhibits proliferation,migration and invasion by targeting LRP6 in papillary thyroid carcinoma[J].Eur Rev Med Pharmaco,2018,22(12):3804-3811.
• [20] KONG F,LI L,WANG C,et al.miR-381-3p suppresses biological characteristics of cancer in head-neck squamous cell carcinoma cells by targeting nuclear autoantigenic sperm protein (NASP)[J].Biosci Biotech Bioch,2020,84(4):703-713.