D-葡萄糖同系物作为人类肝癌细胞BEL-7402荧光探针的价值
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摘要
目的评价荧光D-葡萄糖同系物2-NBDG作为肝癌细胞BEL-7402荧光探针的价值。方法肝癌细胞株BEL-7402培养24 h后分为实验组及对照组,其中实验组加入5 mmol/L D-葡萄糖的DMEM培养基,对照组加入不含5 mmol/L D-葡萄糖的DMEM培养基,再将两组各分为两个亚组加入不同浓度的2-NBDG(0.3 mmol/L、1 mmol/L),即共4组:0.3 mmol/L2-NBDG+5 mmol/L D-葡萄糖组、1 mmol/L 2-NBDG+5 mmol/L D-葡萄糖组、0.3 mmol/L 2-NBDG组、1 mmol/L 2-NBDG组。比较各组细胞荧光强度,分析2-NBDG在肝癌细胞BEL-7402中的摄取及荧光成像情况,以及D-葡萄糖对2-NBDG的抑制竞争作用。结果 0.3 mmol/L 2-NBDG组的相对荧光强度为(78.72±3.78)%,低于1 mmol/L 2-NBDG组的(100.00±8.23)%(P<0.05)。0.3 mmol/L 2-NBDG+5 mmol/L D-葡萄糖组的相对荧光强度为(20.67±1.46)%,明显低于0.3 mmol/L 2-NBDG组的(78.72±3.78)%(P<0.05)。1 mmol/L 2-NBDG+5 mmol/L D-葡萄糖组的相对荧光强度为(22.23±2.01)%,明显低于1 mmol/L 2-NBDG组的(100.00±8.23)%(P<0.05)。结论 2-NBDG可被肝癌细胞BEL-7402快速摄取,并可通过荧光强度的方式量化其摄取程度,D-葡萄糖可竞争性抑制肝癌细胞BEL-7402对2-NBDG摄取。2-NBDG可以作为人源肝癌细胞株BEL-7402的检测荧光探针。
Objective To evaluate the value of D-glucose analog( 2-NBDG) as a fluorescent probe for BEL-7402 human hepatocarcinoma cells. Methods BEL-7402 human hepatocarcinoma cell strains were divided into experimental group with 5 mmol / L D-glucose and control group without 5 mmol / L D-glucose after 24 hours of culture. The two groups were divided into subgroups,and then were added with 2-NBDG with different concentrations( 0. 3 mmol / L and 1 mmol / L). There were 4 groups obtained,including 0. 3 mmol / L 2-NBDG + 5 mmol / L D-glucose group,1 mmol / L2-NBDG + 5 mmol / L D-glucose group,0. 3 mmol / L 2-NBDG group and 1 mmol / L 2-NBDG group. The fluorescence intensities were compared among the 4 groups. The intake and fluorescence imaging of 2-NBDG in BEL-7402 human hepatocarcinoma cells were analyzed. And the competitive antagonism of 2-NBDG by D-glucose was also assessed. Results The relative fluorescence intensity of 0. 3 mmol / L 2-NBDG group was( 78. 72 ±3. 78) %,and was lower than that of 1 mmol/L 2-NBDG group[( 100.00 ±8.23) %]( P <0.05). The relative fluorescence intensity of0. 3 mmol / L 2-NBDG + 5 mmol / L D-glucose group was( 20. 67 ± 1. 46) %,and was significantly lower than that of 0. 3 mmol / L 2-NBDG group[( 78.72 ±3.78) %]( P <0.05). The relative fluorescence intensity of 1 mmol/L 2-NBDG +5 mmol/L D-glucose group was( 22.23 ±2.01) %,and was significantly lower than that of 1 mmol / L 2-NBDG group [( 100. 00 ± 8. 23) % ]( P < 0. 05). Conclusion BEL-7402 human hepatocarcinoma cells can intake 2-NBDG rapidly,and the intake can be assessed quantitatively by the fluorescence intensity. D-glucose might competitively inhibit the intake of 2-NBDG in BEL-7402 hepatocarcinoma cells. 2-NBDG can be taken as a fluorescent probe for BEL-7402 human hepatocarcinoma cells.
Objective To evaluate the value of D-glucose analog( 2-NBDG) as a fluorescent probe for BEL-7402 human hepatocarcinoma cells. Methods BEL-7402 human hepatocarcinoma cell strains were divided into experimental group with 5 mmol / L D-glucose and control group without 5 mmol / L D-glucose after 24 hours of culture. The two groups were divided into subgroups,and then were added with 2-NBDG with different concentrations( 0. 3 mmol / L and 1 mmol / L). There were 4 groups obtained,including 0. 3 mmol / L 2-NBDG + 5 mmol / L D-glucose group,1 mmol / L2-NBDG + 5 mmol / L D-glucose group,0. 3 mmol / L 2-NBDG group and 1 mmol / L 2-NBDG group. The fluorescence intensities were compared among the 4 groups. The intake and fluorescence imaging of 2-NBDG in BEL-7402 human hepatocarcinoma cells were analyzed. And the competitive antagonism of 2-NBDG by D-glucose was also assessed. Results The relative fluorescence intensity of 0. 3 mmol / L 2-NBDG group was( 78. 72 ±3. 78) %,and was lower than that of 1 mmol/L 2-NBDG group[( 100.00 ±8.23) %]( P <0.05). The relative fluorescence intensity of0. 3 mmol / L 2-NBDG + 5 mmol / L D-glucose group was( 20. 67 ± 1. 46) %,and was significantly lower than that of 0. 3 mmol / L 2-NBDG group[( 78.72 ±3.78) %]( P <0.05). The relative fluorescence intensity of 1 mmol/L 2-NBDG +5 mmol/L D-glucose group was( 22.23 ±2.01) %,and was significantly lower than that of 1 mmol / L 2-NBDG group [( 100. 00 ± 8. 23) % ]( P < 0. 05). Conclusion BEL-7402 human hepatocarcinoma cells can intake 2-NBDG rapidly,and the intake can be assessed quantitatively by the fluorescence intensity. D-glucose might competitively inhibit the intake of 2-NBDG in BEL-7402 hepatocarcinoma cells. 2-NBDG can be taken as a fluorescent probe for BEL-7402 human hepatocarcinoma cells.
引文
[1]Cupino AC,Hair CD,Angle JF,et al.Does external beam radiation therapy improve survival following transarterial chemoembolization for unresectable hepatocellular carcinoma?[J].Gastrointest Cancer Res,2012,5(1):13-17.
[2]汪永录,周汉高,顾公望.肝癌研究进展[M].上海:上海科学技术文献出版社,1999:1-72.
[3]Jerusalem G,Hustinx R,Beguin Y,et al.PET scan imaging in oncology[J].Eur J Cancer,2003,39(11):1 525-1 534.
[4]Pauwels EK,Sturm EJ,Bombardieri E,et al.Positron-emission tomography with[18F]fluorodeoxyflucose.Part I.Biochemical uptake mechanism and its implication for clinical studies[J].J Cancer Res Clin Oncol,2000,126(10):549-559.
[5]Warburg O.On the origin of cancer cells[J].Science,1956,125(3 191):309-314.
[6]van Baardwijk A,Dooms C,van Suylen RJ,et al.The maximum uptake of(18)F-deoxyglucose on positron emission tomography scan correlates with survival,hypoxia inducible factor-1alpha and GLUT-1 in non-small cell lung cancer[J].Eur J Cancer,2007,43(9):1 392-1 398.
[7]Yamada K,Nakata M,Horimoto N,et al.Measurement of glucose uptake and intracellular calcium concentration in single,living pancreaticβ-cells[J].J Biol Chem,2000,275(29):22 278-22 283.
[8]Yoshioka K,Takahashi T,Homma T,et al.A novel fluorescent derivative of glucose applicable to the assessment of glucose uptake activity of Escherichia coli[J].Biochim Biophys Acta,1996,1 289(1):5-9.
[9]O'Neil RG,Wu L,Mullani N.Uptake of a fluorescent deoxyglucose analog(2-NBDG)in tumor cells[J].Mol Imaging Biol,2005,7(6):388-392.
[10]Lloyd PG,Hardin CD,Sturek M.Examining glucose transport in single vascular smooth muscle cells with a fluorescent glucose analog[J].Physiol Res,1999,48(6):401-410.
[11]Yoshioka K,Saito M,Oh KB,et al.Intracellular fate of 2-NBDG,a fluorescent probe for glucose uptake activity,in Escherichia coli cells[J].Biosci Biotechnol Biochem,1996,60(11):1 899-1 901.
[12]Yoshioka K,Oh KB,Saito M,et al.Evaluation of 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-D-glucose,a new fluorescent derivative of glucose,for viability assessment of yeast Candida albicans[J].Appl Microbiol Biotechnol,1996,46(4):400-404.
[13]Younes M,Lechago LV,Somoano JR,et al.Wide expression of the human erythrocyte glucose transporter Glut1 in human cancers[J].Cancer Res,1996,56(5):1 164-1 167.
[14]Amann T,Maegdefrau U,Hartmann A,et al.GLUT1 expression is increased in hepatocellular carcinoma and promotes tumorigenesis[J].Am J Pathol,2009,174(4):1 544-1 552.
[15]Yalcin A,Telang S,Clem B,et al.Regulation of glucose metabolism by 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatases in cancer[J].Exp Mol Pathol,2009,86(3):174-179.
[16]Song YS,Lee WW,Chung JH,et al.Correlation between FDG uptake and glucose transporter type 1 expression in neuroendocrine tumors of the lung[J].Lung Cancer,2008,61(1):54-60.
[17]Cao J,Cui S,Li S,et al.Targeted cancer therapy with a 2-deoxyglucose-based adriamycin complex[J].Cancer Res,2013,73(4):1 362-1 372.
[2]汪永录,周汉高,顾公望.肝癌研究进展[M].上海:上海科学技术文献出版社,1999:1-72.
[3]Jerusalem G,Hustinx R,Beguin Y,et al.PET scan imaging in oncology[J].Eur J Cancer,2003,39(11):1 525-1 534.
[4]Pauwels EK,Sturm EJ,Bombardieri E,et al.Positron-emission tomography with[18F]fluorodeoxyflucose.Part I.Biochemical uptake mechanism and its implication for clinical studies[J].J Cancer Res Clin Oncol,2000,126(10):549-559.
[5]Warburg O.On the origin of cancer cells[J].Science,1956,125(3 191):309-314.
[6]van Baardwijk A,Dooms C,van Suylen RJ,et al.The maximum uptake of(18)F-deoxyglucose on positron emission tomography scan correlates with survival,hypoxia inducible factor-1alpha and GLUT-1 in non-small cell lung cancer[J].Eur J Cancer,2007,43(9):1 392-1 398.
[7]Yamada K,Nakata M,Horimoto N,et al.Measurement of glucose uptake and intracellular calcium concentration in single,living pancreaticβ-cells[J].J Biol Chem,2000,275(29):22 278-22 283.
[8]Yoshioka K,Takahashi T,Homma T,et al.A novel fluorescent derivative of glucose applicable to the assessment of glucose uptake activity of Escherichia coli[J].Biochim Biophys Acta,1996,1 289(1):5-9.
[9]O'Neil RG,Wu L,Mullani N.Uptake of a fluorescent deoxyglucose analog(2-NBDG)in tumor cells[J].Mol Imaging Biol,2005,7(6):388-392.
[10]Lloyd PG,Hardin CD,Sturek M.Examining glucose transport in single vascular smooth muscle cells with a fluorescent glucose analog[J].Physiol Res,1999,48(6):401-410.
[11]Yoshioka K,Saito M,Oh KB,et al.Intracellular fate of 2-NBDG,a fluorescent probe for glucose uptake activity,in Escherichia coli cells[J].Biosci Biotechnol Biochem,1996,60(11):1 899-1 901.
[12]Yoshioka K,Oh KB,Saito M,et al.Evaluation of 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-D-glucose,a new fluorescent derivative of glucose,for viability assessment of yeast Candida albicans[J].Appl Microbiol Biotechnol,1996,46(4):400-404.
[13]Younes M,Lechago LV,Somoano JR,et al.Wide expression of the human erythrocyte glucose transporter Glut1 in human cancers[J].Cancer Res,1996,56(5):1 164-1 167.
[14]Amann T,Maegdefrau U,Hartmann A,et al.GLUT1 expression is increased in hepatocellular carcinoma and promotes tumorigenesis[J].Am J Pathol,2009,174(4):1 544-1 552.
[15]Yalcin A,Telang S,Clem B,et al.Regulation of glucose metabolism by 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatases in cancer[J].Exp Mol Pathol,2009,86(3):174-179.
[16]Song YS,Lee WW,Chung JH,et al.Correlation between FDG uptake and glucose transporter type 1 expression in neuroendocrine tumors of the lung[J].Lung Cancer,2008,61(1):54-60.
[17]Cao J,Cui S,Li S,et al.Targeted cancer therapy with a 2-deoxyglucose-based adriamycin complex[J].Cancer Res,2013,73(4):1 362-1 372.