基于毛细管电泳-质谱联用的PTEN缺失诱导代谢重编程研究
|
摘要
前列腺癌是男性常见的恶性肿瘤之一,PTEN缺失普遍存在于前列腺癌中,并诱导肿瘤细胞代谢重编程。PTEN缺失诱导的代谢重编程可能为肿瘤靶向治疗提供信息。本实验基于毛细管电泳-质谱联用技术(CE-MS)对PTEN缺失后前列腺癌细胞DU145及正常前列腺细胞RWPE1代谢变化进行系统性分析。在DU145和RWPE1两种细胞中分别检测到200和214种代谢物,相比对照细胞,PTEN敲除后分别含有28和37种差异代谢物。两种细胞敲除PTEN后,均出现苏糖酸升高,异丁基肉碱、二磷酸腺苷、N-羟乙酰神经氨酸、天冬氨酸、亚牛磺酸下降。DU145敲除PTEN后,特异的代谢变化为L-2-羟戊二酸、甘磷酸胆碱、维生素B1、还原型谷胱甘肽/氧化型谷胱甘肽,且都显著升高,这些代谢物或比值的升高能促进肿瘤细胞的增殖、侵袭及抗化疗。同时,发现肌酸酐、肌肽、乙酰神经氨酸等已报道与癌症诊断、预后相关的标志物受PTEN调控。通过对两种细胞差异代谢物比较,鉴定了PTEN诱导的代谢变化及PTEN与其它肿瘤相关基因共同诱导的代谢变化,为PTEN对代谢通路的调控分子机制研究提供信息。
Prostate cancer is the most frequently occurred cancer in males. Phosphatase and tensin homolog( PTEN) deficiency often occurs in prostate cancer and induces metabolic reprogramming. Metabolic vulnerabilities induced by PTEN deficiency may provide therapeutic targets for cancer therapy. Here,capillary electrophoresis-mass spectrometry( CE-MS) based metabolomics analysis was used for analyzing metabolic changes induced by PTEN deficiency in prostate cancer cell DU145 and normal prostate cell RWPE1. 200 and214 metabolites were detected,and 28 and 37 differential metabolites were authenticated in PTEN knockdowned DU145 and RWPE1 cells compared to their controls,respectively. Threonic acid levels increased,while isobutyrylcarnitine, adenosine diphosphate, N-glycolylneuraminic acid, Asp, hypotaurine levels decreased after PTEN silencing in both cell lines. The specific metabolites changes in DU145 after PTEN silencing were L-2-HG,glycerophosphocholine,thiamine,the ratio of GSH to GSSG,and all of them were increased. These metabolites can promote tumor proliferation,metastasis,and resistance to chemotherapy.Creatinine,carnosine and N-acetylneuraminic acid,which had been reported to be biomarkers of cancer diagnosis and prognosis,were regulated by PTEN deficiency. Metabolites changes induced by PTEN deficiency only or combined effect of PTEN deficiency and other cancer-related genes were identified.
Prostate cancer is the most frequently occurred cancer in males. Phosphatase and tensin homolog( PTEN) deficiency often occurs in prostate cancer and induces metabolic reprogramming. Metabolic vulnerabilities induced by PTEN deficiency may provide therapeutic targets for cancer therapy. Here,capillary electrophoresis-mass spectrometry( CE-MS) based metabolomics analysis was used for analyzing metabolic changes induced by PTEN deficiency in prostate cancer cell DU145 and normal prostate cell RWPE1. 200 and214 metabolites were detected,and 28 and 37 differential metabolites were authenticated in PTEN knockdowned DU145 and RWPE1 cells compared to their controls,respectively. Threonic acid levels increased,while isobutyrylcarnitine, adenosine diphosphate, N-glycolylneuraminic acid, Asp, hypotaurine levels decreased after PTEN silencing in both cell lines. The specific metabolites changes in DU145 after PTEN silencing were L-2-HG,glycerophosphocholine,thiamine,the ratio of GSH to GSSG,and all of them were increased. These metabolites can promote tumor proliferation,metastasis,and resistance to chemotherapy.Creatinine,carnosine and N-acetylneuraminic acid,which had been reported to be biomarkers of cancer diagnosis and prognosis,were regulated by PTEN deficiency. Metabolites changes induced by PTEN deficiency only or combined effect of PTEN deficiency and other cancer-related genes were identified.
引文
1 Torre L A,Bray F,Siegel R L,Ferlay J,Lortet-Tieulent J,Jemal A. Ca. Cancer J. Clin.,2015,65(2):87-108
2 Gray I C,Phillips S M,Lee S J,Neoptolemos J P,Weissenbach J,Spurr N K. Cancer Res.,1995,55(21):4800-4803
3 Li J,Yen C,Liaw D,Podsypanina K,Bose S,Wang S I,Puc J,Miliaresis C,Rodgers L,McCombie R,Bigner S H,Giovanella B C,Ittmann M,Tycko B,Hibshoosh H,Wigler M H,Parsons R. Science,1997,275(5308):1943-1947
4 Mithal P,Allott E,Gerber L,Reid J,Welbourn W,Tikishvili E,Park J,Younus A,Sangale Z,Lanchbury J S,Stone S,Freedland S J. Int. J. Urol.,2014,21(12):1209-1214
5 Kachaylo E,Tschuor C,Calo N,Borgeaud N,Ungethüm U,Limani P,Piguet AC,Dufour J F,Foti M,Graf R,Clavien P A,Humar B. Hepatology,2017,66(3):908-921
6 Lien E C,Lyssiotis C A,Juvekar A,Hu H,Asara J M,Cantley L C,Toker A. Nat. Cell Biol.,2016,18(5):572-578
7 Naguib A,Mathew G,Reczek C R,Watrud K,Ambrico A,Herzka T,Salas I C,Lee M F,El-Amine N,Zheng W,Di Francesco M E,Marszalek J R,Pappin D J,Chandel N S,Trotman L C. Cell Rep.,2018,23(1):58-67
8 Wishart D S,Feunang Y D,Marcu A,Guo A C,Liang K,Vázquez-Fresno R,Sajed T,Johnson D,Li C,Karu N,Sayeeda Z,Lo E,Assempour N,Berjanskii M,Singhal S,Arndt D,Liang Y,Badran H,Grant J,Serra-Cayuela A,Liu Y,Mandal R,Neveu V,Pon A,Knox C,Wilson M,Manach C,Scalbert A. Nucleic Acids Res.,2018,46(D1):D608-D617
9 Fiehn O. Plant Mol. Biol.,2002,48(1-2):155-171
10 Hanahan D,Weinberg R A. Cell,2011,144(5):646-674
11 Zeng J,Kuang H,Hu C,Shi X,Yan M,Xu L,Wang L,Xu C,Xu G. Environ Sci. Technol.,2013,47(13):7457-7465
12 Lindenburg P W,Haselberg R,Rozing G,Ramautar R. Chromatographia,2015,78(5-6):367-377
13 Gulersonmez M C,Lock S,Hankemeier T,Ramautar R. Electrophoresis,2016,37(7-8):1007-1014
14 Kim J,Hu Z,Cai L,Li K,Choi E,Faubert B,Bezwada D,Rodriguez-Canales J,Villalobos P,Lin Y F,Ni M,Huffman KE,Girard L,Byers L A,Unsal-Kacmaz K,Pe1a C G,Heymach J V,Wauters E,Vansteenkiste J,Castrillon D H,Chen B P C,Wistuba I,Lambrechts D,Xu J,Minna J D,De Berardinis R J. Nature,2017,546(7656):168-172
15 Yun J,Mullarky E,Lu C,Bosch K N,Kavalier A,Rivera K,Roper J,Chio I I,Giannopoulou EG,Rago C,Muley A,Asara J M,Paik J,Elemento O,Chen Z,Pappin D J,Dow LE,Papadopoulos N,Gross S S,Cantley L C. Science,2015,350(6266):1391-1396
16 Chow L M,Baker S J. Cancer Lett.,2006,241(2):184-196
17 Chong J,Soufan O,Li C,Caraus I,Li S,Bourque G,Wishart D S,Xia J. Nucleic Acids Res.,2018,46(W1):W486-W494
18 Atkinson D E,Walton G M. J. Biol. Chem.,1967,242(13):3239-3241
19 Intlekofer A M,Wang B,Liu H,Shah H,Carmona-Fontaine C,Rustenburg A S,Salah S,Gunner M R,Chodera J D,Cross J R,Thompson CB. Nat. Chem. Biol.,2017,13(5):494-500
20 Intlekofer A M,Dematteo R G,Venneti S,Finley L W,Lu C,Judkins A R,Rustenburg A S,Grinaway P B,Chodera J D,Cross J R,Thompson C B. Cell Metab.,2015,22(2):304-311
21 Kato Y,Ozawa S,Miyamoto C,Maehata Y,Suzuki A,Maeda T,Baba Y. Cancer Cell Int.,2013,13(1):89
22 Muz B,de la Puente P,Azab F,Azab A K. Hypoxia(Auckl),2015,3:83-92
23 Moestue S A,Borgan E,Huuse E M,Lindholm E M,Sitter B,Brresen-Dale A L,Engebraaten O,Maelandsmo G M,Gribbestad I S. BMC Cancer,2010,10:433
24 Giskedegrd G F,Grinde M T,Sitter B,Axelson D E,Lundgren S,Fjsne H E,Dahl S,Gribbestad I S,Bathen T F.J. Proteome Res.,2010,9(2):972-979
25 Stewart J D,Marchan R,Lesjak M S,Lambert J,Hergenroeder R,Ellis J K,Lau C H,Keun H C,Schmitz G,Schiller J,Eibisch M,Hedberg C,Waldmann H,Lausch E,Tanner B,Sehouli J,Sagemueller J,Staude H,Steiner E,Hengstler J G. Proc. Natl. Acad. Sci. USA,2012,109(21):8155-8160
26 Lu'o'ng K V,Nguyên L T. Cancer Genomics Proteomics,2013,10(4):169-185
27 Bansal A,Simon C M. J. Cell Biol.,2018,217(7):2291-2298
28 Weinstein S J,Mackrain K,Stolzenberg-Solomon R Z,Selhub J,Virtamo J,Albanes D. Cancer Epidemiol. Biomarkers Prev.,2009,18(10):2643-2649
29 Iovine B,Oliviero G,Garofalo M,Orefice M,Nocella F,Borbone N,Piccialli V,Centore R,Mazzone M,Piccialli G,Bevilacqua M A. PLo S One,2014,9(5):e96755
30 MathéE A,Patterson A D,Haznadar M,Manna S K,Krausz K W,Bowman E D,Shields P G,Idle J R,Smith P B,Anami K,Kazandjian D G,Hatzakis E,Gonzalez F J,Harris C C. Cancer Res.,2014,74(12):3259-3270
31 Fischer F,Egg G.HNO,1990,38(10):361-363
2 Gray I C,Phillips S M,Lee S J,Neoptolemos J P,Weissenbach J,Spurr N K. Cancer Res.,1995,55(21):4800-4803
3 Li J,Yen C,Liaw D,Podsypanina K,Bose S,Wang S I,Puc J,Miliaresis C,Rodgers L,McCombie R,Bigner S H,Giovanella B C,Ittmann M,Tycko B,Hibshoosh H,Wigler M H,Parsons R. Science,1997,275(5308):1943-1947
4 Mithal P,Allott E,Gerber L,Reid J,Welbourn W,Tikishvili E,Park J,Younus A,Sangale Z,Lanchbury J S,Stone S,Freedland S J. Int. J. Urol.,2014,21(12):1209-1214
5 Kachaylo E,Tschuor C,Calo N,Borgeaud N,Ungethüm U,Limani P,Piguet AC,Dufour J F,Foti M,Graf R,Clavien P A,Humar B. Hepatology,2017,66(3):908-921
6 Lien E C,Lyssiotis C A,Juvekar A,Hu H,Asara J M,Cantley L C,Toker A. Nat. Cell Biol.,2016,18(5):572-578
7 Naguib A,Mathew G,Reczek C R,Watrud K,Ambrico A,Herzka T,Salas I C,Lee M F,El-Amine N,Zheng W,Di Francesco M E,Marszalek J R,Pappin D J,Chandel N S,Trotman L C. Cell Rep.,2018,23(1):58-67
8 Wishart D S,Feunang Y D,Marcu A,Guo A C,Liang K,Vázquez-Fresno R,Sajed T,Johnson D,Li C,Karu N,Sayeeda Z,Lo E,Assempour N,Berjanskii M,Singhal S,Arndt D,Liang Y,Badran H,Grant J,Serra-Cayuela A,Liu Y,Mandal R,Neveu V,Pon A,Knox C,Wilson M,Manach C,Scalbert A. Nucleic Acids Res.,2018,46(D1):D608-D617
9 Fiehn O. Plant Mol. Biol.,2002,48(1-2):155-171
10 Hanahan D,Weinberg R A. Cell,2011,144(5):646-674
11 Zeng J,Kuang H,Hu C,Shi X,Yan M,Xu L,Wang L,Xu C,Xu G. Environ Sci. Technol.,2013,47(13):7457-7465
12 Lindenburg P W,Haselberg R,Rozing G,Ramautar R. Chromatographia,2015,78(5-6):367-377
13 Gulersonmez M C,Lock S,Hankemeier T,Ramautar R. Electrophoresis,2016,37(7-8):1007-1014
14 Kim J,Hu Z,Cai L,Li K,Choi E,Faubert B,Bezwada D,Rodriguez-Canales J,Villalobos P,Lin Y F,Ni M,Huffman KE,Girard L,Byers L A,Unsal-Kacmaz K,Pe1a C G,Heymach J V,Wauters E,Vansteenkiste J,Castrillon D H,Chen B P C,Wistuba I,Lambrechts D,Xu J,Minna J D,De Berardinis R J. Nature,2017,546(7656):168-172
15 Yun J,Mullarky E,Lu C,Bosch K N,Kavalier A,Rivera K,Roper J,Chio I I,Giannopoulou EG,Rago C,Muley A,Asara J M,Paik J,Elemento O,Chen Z,Pappin D J,Dow LE,Papadopoulos N,Gross S S,Cantley L C. Science,2015,350(6266):1391-1396
16 Chow L M,Baker S J. Cancer Lett.,2006,241(2):184-196
17 Chong J,Soufan O,Li C,Caraus I,Li S,Bourque G,Wishart D S,Xia J. Nucleic Acids Res.,2018,46(W1):W486-W494
18 Atkinson D E,Walton G M. J. Biol. Chem.,1967,242(13):3239-3241
19 Intlekofer A M,Wang B,Liu H,Shah H,Carmona-Fontaine C,Rustenburg A S,Salah S,Gunner M R,Chodera J D,Cross J R,Thompson CB. Nat. Chem. Biol.,2017,13(5):494-500
20 Intlekofer A M,Dematteo R G,Venneti S,Finley L W,Lu C,Judkins A R,Rustenburg A S,Grinaway P B,Chodera J D,Cross J R,Thompson C B. Cell Metab.,2015,22(2):304-311
21 Kato Y,Ozawa S,Miyamoto C,Maehata Y,Suzuki A,Maeda T,Baba Y. Cancer Cell Int.,2013,13(1):89
22 Muz B,de la Puente P,Azab F,Azab A K. Hypoxia(Auckl),2015,3:83-92
23 Moestue S A,Borgan E,Huuse E M,Lindholm E M,Sitter B,Brresen-Dale A L,Engebraaten O,Maelandsmo G M,Gribbestad I S. BMC Cancer,2010,10:433
24 Giskedegrd G F,Grinde M T,Sitter B,Axelson D E,Lundgren S,Fjsne H E,Dahl S,Gribbestad I S,Bathen T F.J. Proteome Res.,2010,9(2):972-979
25 Stewart J D,Marchan R,Lesjak M S,Lambert J,Hergenroeder R,Ellis J K,Lau C H,Keun H C,Schmitz G,Schiller J,Eibisch M,Hedberg C,Waldmann H,Lausch E,Tanner B,Sehouli J,Sagemueller J,Staude H,Steiner E,Hengstler J G. Proc. Natl. Acad. Sci. USA,2012,109(21):8155-8160
26 Lu'o'ng K V,Nguyên L T. Cancer Genomics Proteomics,2013,10(4):169-185
27 Bansal A,Simon C M. J. Cell Biol.,2018,217(7):2291-2298
28 Weinstein S J,Mackrain K,Stolzenberg-Solomon R Z,Selhub J,Virtamo J,Albanes D. Cancer Epidemiol. Biomarkers Prev.,2009,18(10):2643-2649
29 Iovine B,Oliviero G,Garofalo M,Orefice M,Nocella F,Borbone N,Piccialli V,Centore R,Mazzone M,Piccialli G,Bevilacqua M A. PLo S One,2014,9(5):e96755
30 MathéE A,Patterson A D,Haznadar M,Manna S K,Krausz K W,Bowman E D,Shields P G,Idle J R,Smith P B,Anami K,Kazandjian D G,Hatzakis E,Gonzalez F J,Harris C C. Cancer Res.,2014,74(12):3259-3270
31 Fischer F,Egg G.HNO,1990,38(10):361-363