摘要
目的研究灵芝乙醇提取物是否通过脾酪氨酸激酶(Syk)影响胰腺癌细胞SW1990的增殖、迁移和侵袭。方法胰腺癌细胞SW1990给予25、50、100μg/ml剂量的灵芝乙醇提取物,采用噻唑蓝(MTT)比色法检测细胞存活,平板克隆测定细胞克隆形成,Transwell小室法评估细胞迁移和侵袭,免疫印迹实验(western blot)分析细胞核相关抗原Ki67(Ki67)、上皮型钙黏附蛋白(E-cadherin)、神经型钙黏附蛋白(N-cadherin)及Syk表达。在SW1990细胞中转染pcDNA3.1-Syk,或转染pcDNA3.1-Syk并给予100μg/ml浓度的灵芝乙醇提取物,利用上述方法考察细胞的增殖、迁移和侵袭变化。结果低、中、高剂量的灵芝乙醇提取物明显降低细胞存活率、克隆形成数、迁移细胞数、侵袭细胞数、Ki67、N-cadherin蛋白表达水平,并显著提高E-cadherin、Syk蛋白表达水平,均呈剂量依赖性(P<0.05)。Syk过表达明显减少细胞的细胞存活率、克隆形成率、迁移细胞数、侵袭细胞数、Ki67、N-cadherin蛋白表达水平,显著增加E-cadherin蛋白表达水平(P<0.05)。Syk过表达后,灵芝乙醇提取物对SW1990细胞存活、克隆形成、迁移、侵袭、Ki67、N-cadherin蛋白表达的抑制作用和其对细胞E-cadherin蛋白表达的促进作用被增强。结论灵芝乙醇提取物通过上调Syk,抑制胰腺癌细胞SW1990的增殖、迁移和侵袭能力。
Objective To investigate the effects of Ganoderma lucidum ethanol extracts on the proliferation,migration and invasion of pancreatic cancer cell SW1990 through spleen tyrosine kinase(Syk)in vitro.Methods The pancreatic cancer cells SW1990 were treated by Ganoderma lucidum ethanol extracts at doses of 25μg/ml,50μg/ml,and 100μg/ml.The cell survival rate was detected by thiazole blue(MTT)colorimetry.The plate clone assay was used to determine the cell clone formation.The migration and invasion were evaluated by transwell chamber method.Western Blot was used to analyze the expressions of cell nucleus related antigen Ki67(Ki67),epithelium type cadherin(E-cadherin),neurotype cadherin(N-cadherin)and Syk.The SW1990 cells were transfected with pcDNA3.1 Syk or pcDNA3.1 Syk and treated by Ganoderma lucidum ethanol extract at a concentration of 100μg/ml,moreover the proliferation,migration and invasion of the cells were examined by the above methods.Results The low-dose,medium-dose and high-dose Ganoderma lucidum ethanol extracts significantly reduced the cell survival rate,clone formation number,migration number,invasion cell number,and the expression levels of Ki67,N-cadherin protein,however,which significantly increased the expression levels of E-cadherin,Syk with a dose-dependent manner(P<0.05).Moreover the Syk overexpression significantly reduced the cell survival rate,clone formation rate,number of migrating cells,invasion cells number,and the expression levels of Ki67 and N-cadherin protein,but,which significantly increased the expression levels of E-cadherin protein(P<0.05).After overexpression of Syk,the inhibitory effects of Ganoderma lucidum ethanol extract on survival,clone formation,migration,invasion,and the expressions of Ki67 and N-cadherin as well as the promotion effects on E-cadherin protein expression were enhanced.Conclusion The Ganoderma lucidum ethanol extracts can inhibit the proliferation,migration and invasion of pancreatic cancer cells SW1990 by up-regulating the expression of Syk.
作者
王海
徐幸幸
马海
彭勇
WANG Hai;XU Xingxing;MA Hai(Department of Genneral Surgery,Nanchong Central Hospital,Sichuang,Nanchong 637000,China;不详)
出处
《河北医药》
CAS
2021年第16期2427-2431,共5页
Hebei Medical Journal
关键词
灵芝
乙醇提取物
胰腺癌
SYK
增殖
迁移
侵袭
ganoderma lucidum
ethanol extract
pancreatic cancer
Syk
proliferation
migration
invasion
