抑制FoxM1基因表达对甲状腺乳头状癌TPC-1细胞糖酵解途径的影响
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摘要
目的探讨抑制FoxM1基因表达对甲状腺乳头状癌(PTC)细胞葡萄糖摄取和乳酸产量的影响及FoxM1与PTC细胞糖酵解途径的相关性。方法用慢病毒rLv-hFoxM1转染PTC TPC-1细胞,检测转染后的TPC-1细胞中Fox M1 mRNA及蛋白表达水平的变化;应用噻唑蓝(MTT)比色法检测抑制FoxM1基因对TPC-1细胞活性的影响;检测转染慢病毒rLv-hFoxM1后的TPC-1细胞在葡萄糖摄取量和乳酸产量方面的变化以及糖酵解途径关键酶己糖激酶1(HK1)、己糖激酶2(HK2),葡萄糖转运体蛋白1(Glut1)含量的变化。结果在TPC-1细胞中,慢病毒rLv-hFoxM1转染可明显抑制TPC-1细胞中FoxM1基因mRNA及蛋白水平的表达,且转染慢病毒rLv-hFoxM1后TPC-1细胞的活性明显下降(P<0.05),随着FoxM1基因表达降低,TPC-1细胞的葡萄糖摄取量及乳酸产量均明显下降(P<0.01),抑制FoxM1基因后,其葡萄糖转运蛋白(Glut1)、己糖激酶1(HK1)和己糖激酶(HK2)含量明显降低。结论抑制FoxM1基因的表达能降低PTC细胞的糖酵解水平。
Objective To investigate the effect of inhibiting FoxM1 gene expression on glucose uptake and lactic acid production in papillary thyroid carcinoma(PTC) cells, and to explore the correlation between FoxM1 and PTC cell glycolysis pathway. Methods The expression of FoxM1 mRNA and protein in PTC TPC-1 cells were detected after lentiviral rLv-hFoxM1 transfection. The effect of inhibiting FoxM1 gene expression on TPC-1 cell viability was detected by MTT method. The changes of glucose intake and lactic acid production in TPC-1 cells transfected with lentiviral rLv-hFoxM1 and the content of the key enzymes of hexokinase isoform 1(HK1), hexokinase isoform 2(HK2) and glucose transporter 1(Glut1) were detected by Western blot. Results In TPC-1 cells, lentiviral rLv-hFoxM1 transfection significantly inhibited the expression of FoxM1 mRNA and protein in TPC-1 cells and the activity of TPC-1 cells was significantly decreased(P<0.05). After the FoxM1 gene was inhibited, the glucose intake and the lactic acid production of TPC-1 cells were significantly decreased(P<0.01). The levels of Glut1, HK1 and HK2 were also significantly decreased(P<0.01). Conclusion Inhibition of FoxM1 gene expression could reduce the level of glycolysis in PTC cells.
Objective To investigate the effect of inhibiting FoxM1 gene expression on glucose uptake and lactic acid production in papillary thyroid carcinoma(PTC) cells, and to explore the correlation between FoxM1 and PTC cell glycolysis pathway. Methods The expression of FoxM1 mRNA and protein in PTC TPC-1 cells were detected after lentiviral rLv-hFoxM1 transfection. The effect of inhibiting FoxM1 gene expression on TPC-1 cell viability was detected by MTT method. The changes of glucose intake and lactic acid production in TPC-1 cells transfected with lentiviral rLv-hFoxM1 and the content of the key enzymes of hexokinase isoform 1(HK1), hexokinase isoform 2(HK2) and glucose transporter 1(Glut1) were detected by Western blot. Results In TPC-1 cells, lentiviral rLv-hFoxM1 transfection significantly inhibited the expression of FoxM1 mRNA and protein in TPC-1 cells and the activity of TPC-1 cells was significantly decreased(P<0.05). After the FoxM1 gene was inhibited, the glucose intake and the lactic acid production of TPC-1 cells were significantly decreased(P<0.01). The levels of Glut1, HK1 and HK2 were also significantly decreased(P<0.01). Conclusion Inhibition of FoxM1 gene expression could reduce the level of glycolysis in PTC cells.
引文
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[19]Chaika NV,Yu F,Purohit V,et al.Differential expression of metabolic genes in tumor and stromal components of primary and metastatic loci in pancreatic adenocarcinoma[J].PLoS One,2012,7(3):e32996.
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[22]Park HJ,Carr JR,Wang Z,et al.FoxM1,a critical regulator of oxidative stress during oncogenesis[J].EMBO J,2009,28(19):2908-18.
[23]Xu J,Huang Z,Lin L,et al.miRNA-130b is required for the ERK/FOXM1 pathway activation-mediated protective effects of isosorbide dinitrate against mesenchymal stem cell senescence induced by high glucose[J].Int J Mol Med,2015,35(1):59-71.
[24]Shang R,Pu M,Li Y,et al.FOXM1 regulates glycolysis in hepatocellular carcinoma by transactivating glucose transporter 1expression[J].Oncol Rep,2017,37(4):2261-9.
[25]Wang Y,Yun Y,Wu B,et al.FOXM1 promotes reprogramming of glucose metabolism in epithelial ovarian cancer cells via activation of GLUT1 and HK2 transcription[J].Oncotarget,2016,7(30):47985-97.
[26]Cui J,Shi M,Xie D,et al.FOXM1 promotes the warburg effect and pancreatic cancer progression via transactivation of LDHAexpression[J].Clin Cancer Res,2014,20(10):2595-606.
[2]Koppenol WH,Bounds PL,Dang CV.Otto Warburg's contributions to current concepts of cancer metabolism[J].Nat Rev Cancer,2011,11(5):325-37.
[3]Vander Heiden MG,Cantley LC,Thompson CB.Understanding the Warburg effect:the metabolic requirements of cell proliferation[J].Science,2009,324(5930):1029-33.
[4]Ying H,Kimmelman AC,Lyssiotis CA,et al.Oncogenic Kras maintains pancreatic tumors through regulation of anabolic glucose metabolism[J].Cell,2012,149(3):656-70.
[5]Hanahan D,Weinberg RA.Hallmarks of cancer:the next generation[J].Cell,2011,144(5):646-74.
[6]袁浩,闵志均,陈进宏,等.甲状腺乳头状癌TPC-1细胞中FoxM1的表达对Ras及CDK1的影响[J].中国普外基础与临床杂志,2016,23(2):172-6.[Yuan H,Min ZJ,Chen JH,et al.Influence of FoxM1 expression on Ras and CDK1 expressions in thyroid papillary carcinoma TPC-1 cells[J].Zhongguo Pu Wai Ji Chu Yu Lin Chuang Za Zhi,2016,23(2):172-6.]
[7]Millour J,Constantinidou D,Stavropoulou AV,et al.FOXM1 is a transcriptional target of ERalpha and has a critical role in breast cancer endocrine sensitivity and resistance[J].Oncogene,2010,29(20):2983-95.
[8]Madureira PA,Varshochi R,Constantinidou D,et al.The Forkhead box M1 protein regulates the transcription of the estrogen receptor alpha in breast cancer cells[J].J Biol Chem,2006,281(35):25167-76.
[9]Li Q,Zhang N,Jia Z,et al.Critical role and regulation of transcription factor FoxM1 in human gastric cancer angiogenesis and progression[J].Cancer Res,2009,69(8):3501-9.
[10]Pandit B,Halasi M,Gartel AL.p53 negatively regulates expression of FoxM1[J].Cell Cycle,2009,8(20):3425-7.
[11]Ahmed M,Uddin S,Hussain AR,et al.FoxM1 and its association with matrix metalloproteinases(MMP)signaling pathway in papillary thyroid carcinoma[J].J Clin Endocrinol Metab,2012,97(1):E1-E13.
[12]Bellelli R,Castellone MD,Garcia-Rostan G,et al.FOXM1 is a molecular determinant of the mitogenic and invasive phenotype of anaplastic thyroid carcinoma[J].Endocr Relat Cancer,2012,19(5):695-710.
[13]袁浩,闵志均,陈进宏,等.FoxMl基因表达与甲状腺乳头状癌TPC-1细胞活性及侵袭性关系的研究[J].中华内分泌外科杂志,2014,8(1):33-7.[Yuan H,Min ZJ,Chen JH,et al.Correlation of FoxM1 with the activity and invasiveness ofpapillary thyroid carcinoma cell TPC-1[J].Zhonghua Nei Fen Mi Wai Ke Za Zhi,2014,8(1):33-7.]
[14]Lynch TP,Ferrer CM,Jackson SR,et al.Critical role of OLinkedβ-N-acetylglucosamine transferase in prostate cancer invasion,angiogenesis,and metastasis[J].J Biol Chem,2012,287(14):11070-81.
[15]Xia LM,Huang WJ,Wang B,et al.Transcriptional up-regulation of FoxM1 in response to hypoxia is mediated by HIF-1[J].J Cell Biochem,2009,106(2):247-56.
[16]Brizel DM,Schroeder T,Scher RL,et al.Elevated tumor lactate concentrations predict for an increased risk of metastases in headand-neck cancer[J].Int J Radiat Oncol Biol Phys,2001,51(2):349-53.
[17]Walenta S,Wetterling M,Lehrke M,et al.High lactate levels predict likelihood of metastases,tumor recurrence,and restricted patient survival in human cervical cancers[J].Cancer Res,2000,60(4):916-21.
[18]Martinez-Outschoorn UE,Prisco M,Ertel A,et al.Ketones and lactate increase cancer cell“stemness,”driving recurrence,metastasis and poor clinical outcome in breast cancer:achieving personalized medicine via Metabolo-Genomics[J].Cell Cycle,2011,10(8):1271-86.
[19]Chaika NV,Yu F,Purohit V,et al.Differential expression of metabolic genes in tumor and stromal components of primary and metastatic loci in pancreatic adenocarcinoma[J].PLoS One,2012,7(3):e32996.
[20]Coelho RG,Cazarin JM,Cavalcanti de Albuquerque JP,et al.Differential glycolytic profile and Warburg effect in papillary thyroid carcinoma cell lines[J].Oncol Rep,2016,36(6):3673-81.
[21]Kim BH,Kim SJ,Kim K,et al.High metabolic tumor volume and total lesion glycolysis are associated with lateral lymph node metastasis in patients with incidentally detected thyroid carcinoma[J].Ann Nucl Med,2015,29(8):721-9.
[22]Park HJ,Carr JR,Wang Z,et al.FoxM1,a critical regulator of oxidative stress during oncogenesis[J].EMBO J,2009,28(19):2908-18.
[23]Xu J,Huang Z,Lin L,et al.miRNA-130b is required for the ERK/FOXM1 pathway activation-mediated protective effects of isosorbide dinitrate against mesenchymal stem cell senescence induced by high glucose[J].Int J Mol Med,2015,35(1):59-71.
[24]Shang R,Pu M,Li Y,et al.FOXM1 regulates glycolysis in hepatocellular carcinoma by transactivating glucose transporter 1expression[J].Oncol Rep,2017,37(4):2261-9.
[25]Wang Y,Yun Y,Wu B,et al.FOXM1 promotes reprogramming of glucose metabolism in epithelial ovarian cancer cells via activation of GLUT1 and HK2 transcription[J].Oncotarget,2016,7(30):47985-97.
[26]Cui J,Shi M,Xie D,et al.FOXM1 promotes the warburg effect and pancreatic cancer progression via transactivation of LDHAexpression[J].Clin Cancer Res,2014,20(10):2595-606.