Background: We verified: (1) whether pancreatic cancer (PC) cell lines (MIAPaCa2, CAPAN-1, PANC-1, BXPC3) conditioned media (CM) alter glucose metabolism of mouse myoblasts C2C12 (MYO); (2) the gene expression profile of control and CM MYO using a microarray experiment with a platform of 5,000 skeletal muscle cDNA. Methods: MYO were incubated with control or CM for 6, 24 and 48 hrs. For the microarray experiments control and CAPAN-1 CM MYO were used to obtain total RNA, extracted after 16 and 26 hrs: 15g were labelled with Cy3 and Cy5 fluorochromes by direct incorporation (RT). Results: After 24 hrs lactate increased in all CM MYO [control MYO 0.48 0.07 mmol/L, mean increment SEM; CAPAN-1- CM 1.10 0.07 (t 6.19, p 0.001); PANC-1-CM 0.8 0.18 (t 4.48, p 0.01); MIAPaCa 2-CM 1.2 0.13 (t 3.97, p 0.01); BXPC3-CM 1.16 0.13 (t 3.16, p 0.05)]. HK1, GSK3A, PYGM, PKM2, ENO3, ALDOA, GAPDH, PFKM, PDH, IDH2,3,3G, succinylCoA synthetase, SDHD and MDH2 expression did not vary. The expression level at 16 or 26 hrs in comparison with baseline control MYO gave the following results: 23 genes were overexpressed, 8 exerting known biological functions: RPS16, fibrillarin, APOBEC2, BCL2-associated athanogene, ALY, PXR1, PAFAH1B1 and cathepsin G; 24 genes were downregulated, 9 exerting known biological functions: RPS12, thymosin beta 10, troponin T1, IGF2R, RPL14, RPL8, PET112-like, actin alpha and sorcin. The expression levels found in 16 or 26 hrs CM MYO, in comparison with that of 16 and 26 hrs control MYO, was as follow: a total of 43 genes were overexpressed in CM MYO (among which IDH3B, RPL22, RPS3A, RPS21, propionyl CoA carboxylase), and 22 were underexpressed (among which VAMP5). Conclusions: One or more PC compound/s induces MYO production of lactate, through a mechanism independent from genes involved in glycolysis or glycogen synthesis. PC CM enhances the expression of proteolytic enzymes, as cathepsin G, and reduces the expression of ribosomal proteins, possibly favouring protein degradation with respect to synthesis, thus underlying cancer cachexia. A downregulation of the expression of IGF2R, which is associated with GLUT4, and of VAMP5 might interfere with glucose transport when MYO conditioning persists.

Expression profile of pancreatic cancer cell conditioned myoblast: a 5000 muscle genes microarray analysis

GRECO, ELIANA;ZAMBON, CARLO-FEDERICO;BASSO, DANIELA;FOGAR, PAOLA;VALERIO, ANNA CANDIDA;BELLIN, MILENA;ROMUALDI, CHIARA;LANFRANCHI, GEROLAMO;PEDRAZZOLI, SERGIO;PLEBANI, MARIO
2004

Abstract

Background: We verified: (1) whether pancreatic cancer (PC) cell lines (MIAPaCa2, CAPAN-1, PANC-1, BXPC3) conditioned media (CM) alter glucose metabolism of mouse myoblasts C2C12 (MYO); (2) the gene expression profile of control and CM MYO using a microarray experiment with a platform of 5,000 skeletal muscle cDNA. Methods: MYO were incubated with control or CM for 6, 24 and 48 hrs. For the microarray experiments control and CAPAN-1 CM MYO were used to obtain total RNA, extracted after 16 and 26 hrs: 15g were labelled with Cy3 and Cy5 fluorochromes by direct incorporation (RT). Results: After 24 hrs lactate increased in all CM MYO [control MYO 0.48 0.07 mmol/L, mean increment SEM; CAPAN-1- CM 1.10 0.07 (t 6.19, p 0.001); PANC-1-CM 0.8 0.18 (t 4.48, p 0.01); MIAPaCa 2-CM 1.2 0.13 (t 3.97, p 0.01); BXPC3-CM 1.16 0.13 (t 3.16, p 0.05)]. HK1, GSK3A, PYGM, PKM2, ENO3, ALDOA, GAPDH, PFKM, PDH, IDH2,3,3G, succinylCoA synthetase, SDHD and MDH2 expression did not vary. The expression level at 16 or 26 hrs in comparison with baseline control MYO gave the following results: 23 genes were overexpressed, 8 exerting known biological functions: RPS16, fibrillarin, APOBEC2, BCL2-associated athanogene, ALY, PXR1, PAFAH1B1 and cathepsin G; 24 genes were downregulated, 9 exerting known biological functions: RPS12, thymosin beta 10, troponin T1, IGF2R, RPL14, RPL8, PET112-like, actin alpha and sorcin. The expression levels found in 16 or 26 hrs CM MYO, in comparison with that of 16 and 26 hrs control MYO, was as follow: a total of 43 genes were overexpressed in CM MYO (among which IDH3B, RPL22, RPS3A, RPS21, propionyl CoA carboxylase), and 22 were underexpressed (among which VAMP5). Conclusions: One or more PC compound/s induces MYO production of lactate, through a mechanism independent from genes involved in glycolysis or glycogen synthesis. PC CM enhances the expression of proteolytic enzymes, as cathepsin G, and reduces the expression of ribosomal proteins, possibly favouring protein degradation with respect to synthesis, thus underlying cancer cachexia. A downregulation of the expression of IGF2R, which is associated with GLUT4, and of VAMP5 might interfere with glucose transport when MYO conditioning persists.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11577/2439884
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