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目的:研究二甲双胍对人胰腺癌细胞株PANC-1增殖、凋亡及侵袭的影响,并初步探讨其可能机制。方法:体外培养人胰腺癌细胞株PANC-1,给予不同浓度二甲双胍进行干预作为实验组,无药物组作为对照组。MTT法检测二甲双胍对PANC-1细胞存活率的影响;流式细胞仪检测二甲双胍对PANC-1细胞周期及细胞凋亡的影响;Transwell小室侵袭实验观察细胞侵袭能力;RT-PCR检测Bax、Bcl-2、Caspase-3、Cyclin D1、MMP-2和MMP-9mRNA的表达。结果:与对照组相比,用药组细胞增殖活性明显下降,F值分别为70.318、327.201和654.196,P<0.01,且呈时间-浓度依赖性。流式细胞仪检测细胞周期结果显示,随着二甲双胍浓度的增加,G0/G1期细胞所占百分率逐渐增加,S期和G2/M期细胞所占百分率逐渐下降,F值分别为208.365、195.308和48.87,P<0.01;流式细胞仪分析对照组与实验组早期和晚期细胞凋亡率差异无统计学意义,F值分别为0.123和0.298,P>0.05。侵袭实验结果显示,随着二甲双胍浓度的增加,细胞侵袭能力逐渐下降,F=66.131,P<0.01。RT-PCR示,二甲双胍干预组Cyclin D1、MMP-2和MMP-9mRNA表达明显降低,t值分别为6.789、13.452和25.72,P<0.01;Bax、Bcl-2和Caspase-3mRNA与对照组相比差异无统计学意义,t值分别为-0.683、-1.567和0.78,P>0.05。结论:二甲双胍能显著抑制人胰腺癌细胞株PANC-1的增殖和侵袭,机制主要与其阻滞细胞周期以及影响相关基因表达有关;二甲双胍对PANC-1细胞的凋亡无明显诱导作用。
Objective: To study the effect of metformin on the proliferation, apoptosis and invasion of human pancreatic cancer cell line PANC-1 and to explore its possible mechanism. Methods: Human pancreatic cancer cell line PANC-1 was cultured in vitro. Metformin was given as experimental group and no drug group as control group. The effect of metformin on the cell cycle and apoptosis of PANC-1 cells was detected by MTT assay, the cell invasion ability was observed by Transwell assay, the expression of Bax and Bcl- 2, Caspase-3, Cyclin D1, MMP-2 and MMP-9 mRNA expression. Results: Compared with the control group, the cell proliferation activity of the treated group was significantly decreased, with F values of 70.318, 327.201 and 654.196, respectively, P <0.01, in a time-and concentration-dependent manner. The results of flow cytometry showed that the percentage of cells in G0 / G1 phase increased gradually with the increase of metformin concentration, the percentage of cells in S phase and G2 / M phase decreased gradually, the F values were 208.365 and 1955.308 and 48.87, P <0.01). Flow cytometry analysis showed that there was no significant difference between the early and late apoptosis rates in the control group and the experimental group (F = 0.123 and 0.298, P> 0.05). The results of invasion assay showed that with the increase of metformin concentration, cell invasion ability decreased gradually, F = 66.131, P <0.01. RT-PCR showed that the expression of Cyclin D1, MMP-2 and MMP-9 mRNA in metformin group were significantly decreased, t values were 6.789, 13.452 and 25.72, respectively, P <0.01; Bax, Bcl-2 and Caspase-3 mRNA The difference was not statistically significant, t values were -0.683, -1.567 and 0.78, P> 0.05. CONCLUSION: Metformin can significantly inhibit the proliferation and invasion of human pancreatic cancer cell line PANC-1. The mechanism is mainly related to the arrest of cell cycle and the related gene expression. Metformin has no obvious effect on the apoptosis of PANC-1 cells.