新疆紫草中乙酰阿卡宁对人黑色素瘤A375细胞增殖、迁移及侵袭的影响  

Effects of acetylalkannin from Arnebia euchroma on proliferation,migration,and invasion of human melanoma A375 cells

作　　者：康莹莹 钱乾 杨雅 阳莹[1] 许芳 李敏[1] 李建光 KANG Ying-ying;QIAN Qian;YANG Ya;YANG Ying;XU Fang;LI Min;LI Jian-guang(College of Pharmacy of Xinjiang Medical University,Urumqi 830011,China;Xinjiang University of Science and Technology,Korla 841000,China)

机构地区：[1]新疆医科大学药学院,新疆乌鲁木齐830011 [2]新疆科技学院,新疆库尔勒841000

出　　处：《中国中药杂志》2023年第18期5049-5055,共7页China Journal of Chinese Materia Medica

基　　金：新疆天然药物活性组分与释药技术重点实验室项目(XJDX1713);新疆维吾尔自治区科技支疆项目(2018E02069);省部共建中亚高发病成因与防治国家重点实验室开放课题项目(SKL-HIDCA-2018-11)。

摘　　要：观察新疆紫草中乙酰阿卡宁对人黑色素瘤A375细胞增殖、迁移和侵袭的影响,同时探讨其可能的作用机制。设空白组以及乙酰阿卡宁低、中、高剂量组(0.5、1.0、2.0μmol·L-1)分别作用于A375细胞。采用噻唑蓝(MTT)法检测细胞增殖情况,细胞划痕及Transwell迁移实验检测细胞迁移能力;Transwell侵袭实验检测细胞侵袭能力;蛋白免疫印迹法(Western blot)检测迁移侵袭相关N-钙黏蛋白(N-cadherin)、波形蛋白(vimentin)、基质金属蛋白酶9(MMP-9)以及Wnt/β-连环蛋白(β-catenin)通路相关Wnt1、轴蛋白2(Axin2)、糖原合成酶激酶3β(GSK-3β)、磷酸化糖原合酶激酶3β(p-GSK-3β)、β-catenin、细胞周期蛋白D1(cyclin D1)、p21蛋白表达水平;实时荧光定量聚合酶链式反应(real-time PCR)检测E-钙黏蛋白(E-cadherin)、基质金属蛋白酶2(MMP-2)、N-cadherin、vimentin、β-catenin、snail-1、CD44 mRNA表达情况。MTT结果显示,与空白组比较,乙酰阿卡宁各剂量组细胞抑制率显著上升(P<0.01)。细胞划痕和Transwell实验结果显示,与空白组比较,乙酰阿卡宁各组细胞迁移、侵袭数量和迁移侵袭率显著降低(P<0.05,P<0.01),横、纵向迁移及侵袭能力明显减弱。Western blot结果显示,与空白组比较,乙酰阿卡宁高剂量组Axin2蛋白表达增加(P<0.05),N-cadherin、vimentin、MMP-9、Wnt1、p-GSK-3β、β-catenin、cyclin D1、p21蛋白表达均减少(P<0.05,P<0.01),GSK-3β蛋白表达无显著变化;PCR结果显示,MMP-2、N-cadherin、vimentin、β-catenin、snail-1、CD44 mRNA表达总体呈下降趋势(P<0.01),E-cadherin mRNA的表达升高(P<0.01)。乙酰阿卡宁可抑制人黑色素瘤A375细胞的增殖、迁移及侵袭,作用机制可能与调控Wnt/β-catenin信号通路有关。This study aimed to explore the effects and mechanism of acetylalkannin from Arnebia euchroma on the proliferation,migration,and invasion of human melanoma A375 cells.A375 cells were divided into a blank group,and low-,medium-,and high-dose acetylalkannin groups(0.5,1.0,and 2.0μmol·L-1).The MTT assay was used to detect cell proliferation.Cell scratch and transwell migration assays were used to detect cell migration ability,and the transwell invasion assay was used to detect cell invasion ability.Western blot was used to detect the protein expression of migration and invasion-related N-cadherin,vimentin,matrix metalloproteinase-9(MMP-9),and Wnt/β-catenin pathway-related Wnt1,Axin2,glycogen synthase kinase-3β(GSK-3β),phosphorylated GSK-3(p-GSK-3β),β-catenin,cell cycle protein D1(Cyclin D1),and p21.Real-time fluorescence-based quantitative polymerase chain reaction(real-time PCR)was used to detect the mRNA expression of E-cadherin,matrix metalloproteinase-2(MMP-2),N-cadherin,vimentin,β-catenin,snail-1,and CD44.MTT results showed that the cell inhibition rates in the acetylalkannin groups significantly increased as compared with that in the blank group(P<0.01).The results of cell scratch and transwell assays showed that compared with the blank group,the acetylalkannin groups showed reduced cell migration and invasion and migration and invasion rates(P<0.05,P<0.01)and weakened horizontal and vertical migration and invasion abilities.Western blot results showed that compared with the blank group,the acetylalkannin groups showed increased expression of Axin2 protein(P<0.05)and decreased expression of N-cadherin,vimentin,MMP-9,Wnt1,p-GSK-3β,β-catenin,Cyclin D1,and p21 proteins(P<0.05,P<0.01).The expression of GSK-3βprotein did not change significantly.PCR results showed that the overall trend of MM P-2,N-cadherin,vimentin,β-catenin,snail-1,and CD44 mRNA expression was down-regulated(P<0.01),and the expression of E-cadherin mRNA increased(P<0.01).Acetylalkannin can inhibit the proliferation,migration,and invas

关 键 词：新疆紫草 乙酰阿卡宁 黑色素瘤A375细胞 迁移 侵袭 

分 类 号：R285[医药卫生—中药学]