苦参碱通过上调膀胱癌细胞株LINC00472抑制细胞生长和转移  

Matrine inhibits cell growth and metastasis by upregulating bladder cancer cell line LINC00472

作　　者：郭巍[1] 宗实[1] 王凯臣[1] Guo Wei;Zong Shi;Wang Kaichen(Department of Urology,China-Japan Union Hosipital,Jilin University,Changchun 130033,China)

机构地区：[1]吉林大学中日联谊医院泌尿外科,长春130033

出　　处：《中华实验外科杂志》2022年第8期1531-1533,共3页Chinese Journal of Experimental Surgery

基　　金：吉林省卫生科研人才专项项目(2021SCZ27)。

摘　　要：目的探究苦参碱(MAT)对T24和5637细胞的作用及其机制。方法采用细胞计数试剂盒(CCK-8)、流式细胞仪、迁移和侵袭手段检测细胞活力、凋亡、迁移和侵袭潜能;此外, 通过转染试验改变LINC00472的表达, 并通过实时荧光定量聚合酶链反应(RT-qPCR)进行测试。蛋白质印迹法(Western blot)检测细胞周期蛋白D1、p53、B细胞淋巴瘤/白血病-2(bcl-2)、bcl-2相关X蛋白(bax)、pro-Caspase-3、Cleaved-Caspase-3、β-肌动蛋白和细胞通路相关蛋白的水平。应用SPSS 18.0统计软件分析。采用t检验或单向方差分析(ANOVA)。结果苦参碱不影响人输尿管上皮永生化细胞(SV-HUC-1)的生长, 在T24细胞和5637细胞中, 使用MAT后细胞活力分别降为72%和55%, 显著低于对照组, 表明MAT促进肿瘤细胞凋亡, 抑制膀胱癌细胞的生存、侵袭和迁移。此外, LINC00472在膀胱癌组织中显著低表达。苦参碱正向调节LINC00472, 用si-00472转染可以部分逆转苦参碱的功效, 细胞周期蛋白D1和bcl-2的水平显著高于对照组(P<0.01), 而p53、bax和Cleaved-Caspase-3显著低于对照组(P<0.01)。苦参碱提高了第10号染色体上缺失与张力蛋白同源的磷酸酯酶基因(PTEN), 但抑制了磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)蛋白活性(P<0.01)。结论苦参碱通过抑制PTEN/PI3K/Akt通路上调LINC00472, 抑制肿瘤细胞生长和转移。Objective In this experiment,the effect and mechanism of matrine(MAT)on T24 and 5637 cells were preliminarily studied.Methods Cell viability,apoptosis and invasion potential were detected by cell counting kit-8(CCK-8),flow cytometry,migration and invasion assays.In addition,the expression of LINC00472 was altered by transfection assay and tested by real-time quantitative polymerase chain reaction(RT-qPCR).Western blotting was used to detect the levels of Cyclin D1,p53,B cell lymphoma/leukemia-2(bcl-2),bcl-2 associated X protein(bax),pro-Caspase-3,cleaved-Caspase-3,β-actin and cell pathway-related proteins.SPSS 18.0 statistical software was used for analysis.P values were calculated using t-test or one-way analysis of variance(ANOVA).Results MAT did not affect the growth of normal bladder cells(SV-HUC-1).In T24 cells and 5637 cells,the cell viability decreased to 72%and 55%respectively after treatment with MAT,which was significantly lower than that of the control group.MAT promoted tumor cell apoptosis and inhibited the survival,invasion and migration of bladder cancer cells.In addition,LINC00472 was significantly underexpressed in bladder cancer tissues.MAT positively regulated LINC00472,and transfection with si-00472 coulc partially reverse the efficacy of MAT.Compared with control group,the levels of cyclin D1 and bcl-2 were significantly enhanced(P<0.01),while the levels of p53,bax and cleaved-Caspase-3 were significantly decreased(P<0.01).MAT increased phosphatase and tensin homologue deleted on chromosome ten(PTEN)but inhibited phosphatidylinositol 3 kinase/protein kinase B(PI3K/Akt)protein activity(P<0.01).Conclusion MAT up-regulates LINC00472 by inhibiting the PTEN/PI3K/Akt pathway,and inhibits tumor cell growth and metastasis.

关 键 词：膀胱癌 苦参碱 第10号染色体上缺失与张力蛋白同源的磷酸酯酶基因/磷脂酰肌醇3-激酶/蛋白激酶B通路 

分 类 号：R737.14[医药卫生—肿瘤]