摘要
目的:探索二甲双胍对胰腺癌BxPC-3细胞恶性表型的影响。方法:以Smad4基因天然缺失型人胰腺癌BxPC-3细胞为对象,采用CCK-8法和流式细胞术分别检测不同剂量二甲双胍(5、10、20 mmol/L)作用24 h后的细胞增殖和凋亡情况,并计算细胞存活率和凋亡率;采用Transwell迁移试验检测不同剂量二甲双胍(10、20 mmol/L)作用24 h后的细胞迁移情况,记录迁移细胞数;采用实时定量聚合酶链反应法和Western blotting法分别检测细胞中钙黏着蛋白E(E-cadherin)、波形蛋白(Vimentin)、补体反应基因32(RGC-32)的mRNA及蛋白表达情况。结果:与对照组和5 mmol/L二甲双胍组比较,10、20 mmol/L二甲双胍组细胞的存活率均显著降低,凋亡率均显著升高,且20 mmol/L二甲双胍组细胞的凋亡率显著高于10 mmol/L二甲双胍组(P<0.05)。与对照组比较,10、20 mmol/L二甲双胍组迁移细胞数均显著减少,且20 mmol/L二甲双胍组显著少于10 mmol/L二甲双胍组(P<0.05);10、20 mmol/L二甲双胍组细胞中E-cadherin mRNA及其蛋白的相对表达量均显著升高,且20 mmol/L二甲双胍组E-cadherin mRNA的相对表达量显著高于10 mmol/L二甲双胍组;10 mmol/L二甲双胍组细胞中Vimentin mRNA,20 mmol/L二甲双胍组细胞中Vimentin mRNA及其蛋白以及10、20 mmol/L二甲双胍组细胞中RGC-32 mRNA及其蛋白的相对表达量均显著降低,且20 mmol/L二甲双胍组Vimentin mRNA及其蛋白、RGC-32 mRNA的相对表达量均显著低于10 mmol/L二甲双胍组(P<0.05或P<0.01)。结论:二甲双胍可剂量依赖性地通过Smad4非依赖性通路抑制BxPC-3细胞的增殖和迁移,促进其凋亡,这可能与胰腺癌细胞上皮间质转化过程以及RGC-32表达受到抑制有关。
OBJECTIVE:To investigate the effects of metformin on malignant phenotype of pancreatic cancer Bx PC-3 cells.METHODS:Using human pancreatic cancer Bx PC-3 cells with natural deletion of Smad4 gene as reaserch objects,CCK-8 assay and flow cytometry were used to detect the proliferation and apoptosis of Bx PC-3 cells after treated with different doses of metformin(5,10,20 mmol/L)for 24 h. The cell survival rate and apoptosis rate were calculated. Transwell assay was used to test the migration of cells after treated with different doses of metformin(10,20 mmol/L)for 24 h. The number of migrating cells was recorded. q RT-PCR and Western blotting assay were performed to determine m RNA and protein expression of E-cadherin,Vimentin and RGC-32 in cells. RESULTS:Compared with control group and 5 mmol/L metformin group,survival rate of cells were decreased significantly in 10,20 mmol/L metformin groups,while apoptosis rate was increased significantly;the apoptosis rate in20 mmol/L metformin group was significantly higher than 10 mmol/L metformin group(P<0.05). Compared with control group,the number of migrating cells was decreased significantly in 10,20 mmol/L metformin groups,and the 20 mmol/L metformin group was significantly lower than 10 mmol/L metformin group(P<0.05). Relative m RNA and protein expression of E-cadherin were increased significantly in 10,20 mmol/L metformin groups,and relative m RNA expression of E-cadherin in 20 mmol/L metformin group was significantly higher than 10 mmol/L metformin group. Relative m RNA expression of Vimentin in 10 mmol/L metformin group,relative m RNA and protein expression of Vimentin in 20 mmol/L metformin group,relative m RNA and protein expression of RGC-32 in 10,20 mmol/L metformin groups were decreased significantly;relative m RNA and protein expression of Vimentin as well as m RNA expression of RGC-32 in 20 mmol/L metformin group were significantly lower than 10 mmol/L metformin group(P<0.05 or P<0.01). CONCLUSIONS:Metformin can inhibit the proliferation and migration of pancreatic cancer cells through smael-independent pathways in a dosedependent manner, and promote their apoptosis, which is associated with the inhibition epithelial-mesenchymal transition and the expression of RGC-32 of pancreatic cancer.
作者
黄志铨
王振文
朱亮
HUANG Zhiquan;WANG Zhenwen;ZHU Liang(Dept.of Gastroenterology,the First Affiliated Hospital of Nanchang University,Nanchang 330006,China)
出处
《中国药房》
CAS
北大核心
2020年第2期202-207,共6页
China Pharmacy
基金
江西省青年科学基金资助项目(No.20171BAB215044)
江西省教育厅科学技术研究项目(No.170006)
江西省卫生计生委科技计划项目(No.20195082)
