摘要
目的探讨夏枯草对胃癌细胞上皮间质化(epithelial mesenchymal transitions,EMT)的抑制作用及其可能的作用机制。方法选取人胃癌细胞株HGC-27,将细胞随机分为对照组及夏枯草低、中、高剂量组(20、40、80μg·mL^(−1)夏枯草提取物)。用四甲基偶氮唑盐[3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazolium bromide,MTT]法检测细胞增殖抑制率;划痕实验检测细胞迁移率;Transwell小室实验检测细胞侵袭能力;反转录·聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)检测miR-155、糖原合成酶激酶-3β(glycogen synthase kinase-3β,GSK-3β)和β连环蛋白(β-catenin)mRNA表达情况;蛋白质印迹法(Western blotting)检测GSK-3β、β-catenin、E-钙黏蛋白(E-cadherin)、神经性钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)和锌指蛋白(Snail)表达情况。结果与对照组比较,夏枯草3组胃癌细胞增殖抑制率、GSK-3βmRNA、GSK-3β和E-cadherin蛋白表达水平升高,细胞迁移率、细胞侵袭数量、miR-155、β-catenin mRNA水平、β-catenin、N-cadherin、Vimentin和Snail蛋白的表达水平降低(P<0.05)。与夏枯草低剂量组比较,夏枯草中剂量组、夏枯草高剂量组胃癌细胞增殖抑制率、GSK-3βmRNA、GSK-3β和E-cadherin蛋白的表达水平升高,细胞迁移率、细胞侵袭数量、miR-155、β-catenin mRNA水平、β-catenin、N-cadherin、Vimentin和Snail蛋白的表达水平降低(P<0.05),夏枯草高剂量组诸项指数变化尤其明显(P<0.05)。结论夏枯草对胃癌细胞增殖、迁移和侵袭有一定的抑制作用,并能有效阻断胃癌细胞发生EMT,可能是通过降低miRNA-155表达水平、抑制Wnt/β-catenin信号通路发挥作用。
Objective To investigate the inhibitory effect of Prunella vulgaris on epithelial mesenchymal transitions(EMT)of gastric cancer cells and its possible mechanism.Methods Human gastric cancer cell line HGC-27 was selected and the cells were randomly divided into control group,low,medium and high doses of Prunella vulgaris groups(20,40 and 80μg·mL^(−1)Prunella vulgaris).The inhibition rate of cell proliferation was detected by 3(-4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazolium bromide(MTT)assay.The cell mobility was detected by scratch test.The invasion ability of cells was detected by Transwell chamber experiment.The levels of miR-155,glycogen synthase kinase-3β(GSK-3β)andβ-catenin mRNA expression were detected by reverse transcription-polymerase chain reaction(RT-PCR).The levels of GSK-3β,β-catenin,E-cadherin,N-cadherin,Vimentin and Snail protein expression were detected by Western blotting.Results Compared with the control group,the inhibition rate of gastric cancer cell proliferation,the levels of GSK-3βmRNA expression,GSK-3βand E-cadherin protein expression were increased,the cell migration rate,the numbers of cell invasion,the levels of miR-155,β-catenin mRNA expression,β-catenin,N-cadherin,Vimentin and Snail protein expression were decreased in low,medium and high doses of Prunella vulgaris groups(P<0.05).Compared with the low dose group of Prunella vulgaris,the proliferation inhibition rate of gastric cancer cells,the expression levels of GSK-3βmRNA,GSK-3βand E-cadherin proteins increased,and the cell migration rate,the number of cell invasions,the mRNA levels of miR-155 andβ-catenin mRNA,β-catenin,N-cadherin,Vimentin and Snail proteins expression increased in the medium dose and high dose groups of Prunella vulgaris(P<0.05).The indexes of high dose of Prunella vulgaris group changed significantly(P<0.05).Conclusion Prunella vulgaris has a certain inhibitory effect on the proliferation,migration and invasion of gastric cancer cells,and can effectively block the EMT of gastric cancer cells,possibly by reducing the expression of mirna-155 and inhibiting Wnt/β-catenin signaling pathway plays a role.
作者
李云川
王刚
李强
李玉庆
LI Yunchuan;WANG Gang;LI Qiang;LI Yuqing(The Second Department of General Surgery,Tangshan Union Medical College Hospital,Tangshan 063000,China)
出处
《西北药学杂志》
CAS
2024年第2期93-98,共6页
Northwest Pharmaceutical Journal
基金
河南省医学科学研究课题计划项目(编号:20210941)。
