We performed a pilot study using human peripheral blood lymphocytes (PBL) as a novel system to identify new biomarkers of dihydrotestosterone (DHT) and insulin-like growth factor-1 (IGF-1) abuse in sport. First, to obtain a gene signature, we treated cultures of lymphocytes from sedentary males with three doses of 0,237 mg/ml DHT, each of whichis 80-foldthe physiological concentration in young adult male serum, at days 0,2 and 4, or with a single dose of 1,25 mg/ml IGF-1, which is 5-fold the physiological concentration in young male serum. We then used the Human Genome U133Plus 2.0 microrray to identify a gene signature related to DHT or IGF-1 administration. Gene expression wasevaluated after 7 and 21 days of DHT treatment, and after 24 h, 72 h and 7 days of IGF-1 treatment. Microrray analysis yielded a list of genes whose expression was altered after DHT or IGF-1 treatment. Among these we selected the genes that are most representative of the pathways associated with skeletal and muscular disorders using the IPA bioinformatics tool. We identified six (IDO1, CXCL13, CCL1, GZBM, VDR and IL2RA) and two (FNI and RAB31) genes that were up-regulated in lymphocytes from sedentary subjects after 7 days of DHT and IGF-1 treatment, respectively. The expression of these genes in lymphocytes from differently trained athletes was either down-regulated or similar to that in lymphocites from sedentary subjects. The finding suggests that up-regulation was due to the drug and not to physical exercise. In conclusion, we demonstrate that PBL can be useful in anti-doping checks, and we describe new biomarkers of DHT and IGF-1 abuse which can be included in the Athlete’s Biological Passport.
DHT AND IGF-1 IN PERIPHERAL BLOOD LYMPHOCYTES: NEW MARKERS FOR THE ATHLETE'S BIOLOGICAL PASSPORT
PARISI A;
2013-01-01
Abstract
We performed a pilot study using human peripheral blood lymphocytes (PBL) as a novel system to identify new biomarkers of dihydrotestosterone (DHT) and insulin-like growth factor-1 (IGF-1) abuse in sport. First, to obtain a gene signature, we treated cultures of lymphocytes from sedentary males with three doses of 0,237 mg/ml DHT, each of whichis 80-foldthe physiological concentration in young adult male serum, at days 0,2 and 4, or with a single dose of 1,25 mg/ml IGF-1, which is 5-fold the physiological concentration in young male serum. We then used the Human Genome U133Plus 2.0 microrray to identify a gene signature related to DHT or IGF-1 administration. Gene expression wasevaluated after 7 and 21 days of DHT treatment, and after 24 h, 72 h and 7 days of IGF-1 treatment. Microrray analysis yielded a list of genes whose expression was altered after DHT or IGF-1 treatment. Among these we selected the genes that are most representative of the pathways associated with skeletal and muscular disorders using the IPA bioinformatics tool. We identified six (IDO1, CXCL13, CCL1, GZBM, VDR and IL2RA) and two (FNI and RAB31) genes that were up-regulated in lymphocytes from sedentary subjects after 7 days of DHT and IGF-1 treatment, respectively. The expression of these genes in lymphocytes from differently trained athletes was either down-regulated or similar to that in lymphocites from sedentary subjects. The finding suggests that up-regulation was due to the drug and not to physical exercise. In conclusion, we demonstrate that PBL can be useful in anti-doping checks, and we describe new biomarkers of DHT and IGF-1 abuse which can be included in the Athlete’s Biological Passport.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
