摘要
目的 探讨片仔癀(PZH)对AsPC-1、PANC-1细胞增殖、迁移及上皮间质转化(EMT)的调控作用.方法 采用MTT实验筛选出PZH溶液干预AsPC-1、PANC-1细胞的最佳浓度为0、0.5、1 mg/mL.转录组测序检测0、1 mg/mL PZH干预AsPC-1细胞的基因表达变化;划痕愈合实验检测AsPC-1、PANC-1细胞的迁移能力;Transwell迁移实验检测AsPC-1、PANC-1细胞的迁移能力;Western blot检测AsPC-1、PANC-1细胞E-钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)、N-钙黏蛋白(N-cadherin)和E盒结合锌指蛋白1(ZEB1)的蛋白表达.结果 ①转录组测序:0、1 mg/mL组在与EMT相关的重要指标中基因表达量具有明显差异(P<0.05).②划痕愈合实验:与0 mg/mL组比较,1 mg/mL组PANC-1细胞12 h划痕损伤率明显降低(P<0.05),1 mg/mL组AsPC-1细胞24 h划痕损伤率明显降低(P<0.05),0.5、1 mg/mL组AsPC-1和PANC-1细胞48 h划痕损伤率明显降低(P<0.05).③Transwell迁移实验:与0 mg/mL组比较,1 mg/mL组AsPC-1和PANC-1细胞相对迁移比值明显降低(P<0.05).④Western blot实验:与0 mg/mL组比较,1 mg/mL组AsPC-1细胞E-cadherin蛋白表达量明显升高(P<0.05),0.5、1 mg/mL组AsPC-1细胞Vimentin蛋白表达量均明显降低(P<0.05);与0 mg/mL组比较,1 mg/mL组PANC-1细胞N-cadherin、ZEB1蛋白表达量均明显降低(P<0.05).结论 PZH明显抑制人胰腺癌细胞的增殖、迁移及EMT,具有抗胰腺癌转移的潜在功效.
Abstract
Objective:To explore the regulatory effect of PZH on the proliferation,migration and epithelial-mesenchymal transi-tion (EMT) of AsPC-1 and PANC-1 cells. Methods:The MTT assay was used to screen the optimal concentrations of PZH solution for intervention in AsPC-1 and PANC-1 cells as 0,0.5,and 1 mg/mL. Transcriptome sequencing was used to detect gene expression changes in AsPC-1 cells intervened with 0,1 mg/mL PZH;wound healing experiment was used to detect the migration ability of AsPC-1 and PANC-1 cells;transwell migration assay was used to detect the migration ability of AsPC-1 and PANC-1 cells;Western blot was used to detect the protein expression of E-cadherin,Vimentin,N-cadherin,and Zinc finger E-box binding homeobox 1 (ZEB1) in AsPC-1 and PANC-1 cells. Results:1) Transcriptome sequencing:There were significant differences in gene expression levels between the 0 and 1 mg/mL groups in important indicators related to EMT. 2) Scratch healing experiment:Compared with the 0 mg/mL group,the scratch damage rate of PANC-1 cells in the 1 mg/mL group significantly reduced after 12 hours (P<0.05),the scratch damage rate of AsPC-1 cells in the 1 mg/mL group significantly reduced after 24 hours (P<0.05),and the scratch damage rates of AsPC-1 and PANC-1 cells in the 0.5 and 1 mg/mL groups significantly reduced after 48 hours (P<0.05). 3) Transwell experi-ment:Compared with the 0 mg/mL group,the relative migration ratio of AsPC-1 and PANC-1 cells in the 1 mg/mL group significantly reduced (P<0.05). 4) Western blot experiment:Compared with the 0 mg/mL group,the protein expression of E-cadherin of AsPC-1 cells in the 1 mg/mL group significantly increased (P<0.05),while the protein expression of Vimentin of AsPC-1 cells in the 0.5 and 1 mg/mL groups significantly reduced (P<0.05);compared with the 0 mg/mL group,the protein expression of N-cadherin and ZEB1 of PANC-1 cells in the 1 mg/mL group significantly reduced (P<0.05). Conclusion:PZH obviously inhibits the proliferation,migra-tion and EMT of human pancreatic cancer cells,which has the potential effect of anti-metastasis in pancreatic cancer.
维普
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