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二甲双胍对胰腺癌BxPC-3细胞恶性表型的影响
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| 篇名： | 二甲双胍对胰腺癌BxPC-3细胞恶性表型的影响 |
| TITLE： | Effects of Metformin on the Malignant Phenotype of Pancreatic Cancer BxPC- 3 Cells |
| 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、20mmol/L）作用24h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、20mmol/L）作用24h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Westernblotting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5mmol/L二甲双胍组比较，10、20mmol/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20mmol/L二甲双胍组细胞的凋亡率显著高于10mmol/L二甲双胍组（P＜0.05）。与对照组比较，10、20mmol/L二甲双胍组迁移细胞数均显著减少，且20mmol/L二甲双胍组显著少于10mmol/L二甲双胍组（P＜0.05）；10、20mmol/L二甲双胍组细胞中E-cadherinmRNA及其蛋白的相对表达量均显著升高，且20mmol/L二甲双胍组E-cadherinmRNA的相对表达量显著高于10mmol/L二甲双胍组；10mmol/L二甲双胍组细胞中VimentinmRNA，20mmol/L二甲双胍组细胞中Vi-mentinmRNA及其蛋白以及10、20mmol/L二甲双胍组细胞中RGC-32mRNA及其蛋白的相对表达量均显著降低，且20mmol/L二甲双胍组VimentinmRNA及其蛋白、RGC-32mRNA的相对表达量均显著低于10mmol/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。 |
| ABSTRACT： | OBJECTIVE：To investigate the effects of metformin on malignant phenotype of pancreatic cancer BxPC- 3 cells. METHODS：Using human pancreatic cancer BxPC- 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 BxPC- 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. qRT-PCR and Western blotting assay were performed to determine mRNA 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 in 20 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 mRNA and protein expression of E-cadherin were increased significantly in 10，20 mmol/L metformin groups ，and relative mRNA expression of E-cadherin in 20 mmol/L metformin group was significantly higher than 10 mmol/L metformin group. Relative mRNA expression of Vimentin in 10 mmol/L metformin group ，relative mRNA and protein expression of Vimentin in 20 mmol/L metformin group ，relative mRNA and protein expression of RGC- 32 in 10，20 mmol/L metformin groups were decreased significantly ；relative mRNA and protein expression of Vimentin as well as mRNA 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 dose- dependent manner ，and promote their apoptosis ，which is associated with the inhibition epithelial- mesenchymal transition and the expression of RGC- 32 of pancreatic cancer. |
| 期刊： | 2020年第31卷第02期 |
| 作者： | 黄志铨，王振文，朱亮 |
| AUTHORS： | HUANG Zhiquan ，WANG Zhenwen ，ZHU Liang |
| 关键字： | 二甲双胍；胰腺癌；BxPC-3细胞；增殖；凋亡；上皮间质转化；补体反应基因32 |
| KEYWORDS： | Metformin；Pancreatic cancer ；BxPC-3 cell； |
| 阅读数： | 365 次 |
| 本月下载数： | 7 次 |
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