二甲双胍对卵巢癌细胞侵袭迁移及MTA1表达的影响
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摘要
目的:体外细胞培养实验检测二甲双胍对卵巢癌细胞SKOV3侵袭迁移能力的影响,以及其对转移相关因子1(MTA1)表达的影响。方法:用不同浓度二甲双胍处理卵巢癌细胞,采用划痕实验和transwell实验检测二甲双胍对细胞侵袭迁移能力的影响,Western blot法检测二甲双胍对细胞中MTA1蛋白表达的影响,qRT-PCR检测二甲双胍对细胞中MTA1 mRNA表达的影响。结果:与对照组(0mmol/L)相比,二甲双胍干预组的细胞迁移速度明显下降(P<0.01);作用24h时,2.5mmol/L与5mmol/L组未见差异(P>0.05);作用48h时,卵巢癌细胞的迁移速度随着二甲双胍浓度的增加而减弱(P<0.01)。随着二甲双胍浓度的增加,穿膜细胞数减少(P<0.01);二甲双胍干预组的MTA1 mRNA及MTA1蛋白表达明显降低(P<0.05),不同药物浓度之间无差异。结论:二甲双胍抑制SKOV3细胞侵袭迁移,抑制作用与浓度呈正相关,其机制可能与抑制MTA1表达有关。
Objective: To investigate the influence of metformin regulation on SKOV3 in migration and invasion of ovarian cancer. Methods: The human ovarian cancer SKOV3 cells were treated with metformin at a dose of 0,2.5,5.0 and 10.0mmol/L,cell migration and invasion were detected by cell scratch test and Transwell chamber assay respectively.The protein expression level of MTA1 was detected by Western blotting,The mRNA expression of MTA1 was evaluated by real-time quantitative PCR.Results: Compared with the control group( 0mmol/L),the cell migration rate of metformin intervention group decreased significantly( P < 0. 01),and no difference was found between 2.5mmol/L and 5mmol/L after 24h( P>0.05). The number of the cells which invaded matrigel was in a dose-dependent manner. Real-time quantitative PCR showed that the mRNA expression level of MTA1 was decreased in metformin treatment groups,significantly different from those in the control group( P < 0. 05). Western blot showed that the protein level of MTA1 was significantly decreased in metformin treatment groups( 2.5,5.0 and10.0mmol/L) when compared with the control group( P < 0. 05). Conclusions: Metformin inhibit invasion and migration of human ovarian cancer cells,this inhibition was in a dose-dependent manner,and may be related to the inhibition of MTA1 expression.
Objective: To investigate the influence of metformin regulation on SKOV3 in migration and invasion of ovarian cancer. Methods: The human ovarian cancer SKOV3 cells were treated with metformin at a dose of 0,2.5,5.0 and 10.0mmol/L,cell migration and invasion were detected by cell scratch test and Transwell chamber assay respectively.The protein expression level of MTA1 was detected by Western blotting,The mRNA expression of MTA1 was evaluated by real-time quantitative PCR.Results: Compared with the control group( 0mmol/L),the cell migration rate of metformin intervention group decreased significantly( P < 0. 01),and no difference was found between 2.5mmol/L and 5mmol/L after 24h( P>0.05). The number of the cells which invaded matrigel was in a dose-dependent manner. Real-time quantitative PCR showed that the mRNA expression level of MTA1 was decreased in metformin treatment groups,significantly different from those in the control group( P < 0. 05). Western blot showed that the protein level of MTA1 was significantly decreased in metformin treatment groups( 2.5,5.0 and10.0mmol/L) when compared with the control group( P < 0. 05). Conclusions: Metformin inhibit invasion and migration of human ovarian cancer cells,this inhibition was in a dose-dependent manner,and may be related to the inhibition of MTA1 expression.
引文
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[2]Currie CJ,Poole CD,Jenkins-Jones S,et al.Mortality after incident cancer in people with and without type 2 diabetes:impact of metformin on survival[J].Cancer Med,2012,35(2):299-304
[3]Cheng K,Hao M.Metformin inhibits TGF-beta1-induced epithelial-to-mesenchymal transition via PKM2 relative-m TOR/p70s6k signaling pathway in cervical carcinoma Cells[J].Int J Mol Sci,2016,17(12).pii:E2000
[4]Gwak H,Kim Y,An H,et al.Metformin induces degradation of cyclin D1 via AMPK/GSK3beta axis in ovarian cancer[J].Mol Carcinog,2017,56(2):349-358
[5]Kasznicki J,Sliwinska A,Drzewoski J.Metformin in cancer prevention and therapy[J].Ann Transl Med,2014,2(6):57
[6]Moon HS,Kim B,Gwak H,et al.Autophagy and protein kinase RNA-like endoplasmic reticulum kinase(PERK)/eukaryotic initiation factor 2 alpha kinase(e IF2alpha)pathway protect ovarian cancer cells from metformin-induced apoptosis[J].Mol Carcinog,2016,55(4):346-356
[7]Lu Y,Wei C,Xi Z.Curcumin suppresses proliferation and invasion in non-small cell lung cancer by modulation of MTA1-mediated Wnt/beta-catenin pathway[J].In Vitro Cell Dev Biol Anim,2014,50(9):840-850
[8]Weng W,Yin J,Zhang Y,et al.Metastasis-associated protein 1promotes tumor invasion by downregulation of E-cadherin[J].Int J Oncol,2014,44(3):812-818
[9]Wu B,Li S,Sheng L,et al.Metformin inhibits the development and metastasis of ovarian cancer[J].Oncol Rep,2012,28(3):903-908
[10]Gotlieb WH,Saumet J,Beauchamp MC,et al.In vitro metformin anti-neoplastic activity in epithelial ovarian cancer[J].Gynecol Oncol,2008,110(2):246-250
[11]Zhou H,Xu X,Xun Q,et al.microRNA-30c negatively regulates endometrial cancer cells by targeting metastasis-associated gene-1[J].Oncol Rep,2012,27(3):807-812
[12]Wu W,Yang JL,Wang YL,et al.Reversal of multidrug resistance of hepatocellular carcinoma cells by metformin through inhibiting NF-κB gene transcription[J].World J Hepatol,2016,8(23):985-993