cd73 inhibitor apcp  (Tocris)

Journal: Journal of Cancer

Article Title: CD73 is a hypoxia-responsive gene and promotes the Warburg effect of human gastric cancer cells dependent on its enzyme activity

doi: 10.7150/jca.62387

Figure Lengend Snippet: CD73 is induced by hypoxia in gastric cancer. ( A ) Correlation analysis of CD73 expression and hypoxia gene signature in gastric cancer; data were obtained from TCGA cohort. ( B ) Western blotting analysis of CD73 expression in gastric cancer cell lines and GES1 cell line. ( C ) BGC-823 cells were cultured under hypoxia (1% O 2 ) and normoxia (20% O 2 ) for 12 or 24 h, followed by detection of CD73 expression with western blotting. ( D ) BGC-823 cells were cultured with the treatment of 100 μM CoCl 2 or not for 12 or 24 h, followed by detection of CD73 expression with western blotting. ( E ) Under hypoxic culture condition (1% O 2 ), CD73 expression in BGC-823 cells in the presence or absence of HIF1α knockdown was analyzed by western blotting. ( F ) Chromatin immunoprecipitation-PCR validation of the regulatory role of HIF1α in CD73 expression. Input and ChIP cycle threshold (Ct) values were normalized separately to control as 1. ** P < 0.01.

Article Snippet: APCP (CD73 inhibitor) and adenosine were acquired from Sigma (Shanghai, China).

Techniques: Expressing, Western Blot, Cell Culture, Chromatin Immunoprecipitation

Journal: Journal of Cancer

Article Title: CD73 is a hypoxia-responsive gene and promotes the Warburg effect of human gastric cancer cells dependent on its enzyme activity

doi: 10.7150/jca.62387

Figure Lengend Snippet: CD73 regulates the Warburg effect in gastric cancer. ( A ) The knockdown efficiency of two independent shRNAs against CD73 in HGC-27 and SGC-7901 cells was analyzed by western blotting analysis. ( B-D ) Effect of CD73 knockdown on the glucose uptake (B, n = 3), lactate release (C, n = 3), and expression of glycolytic genes (D, n = 3) in HGC-27 and SGC-7901 cells. ( E ) The overexpression efficiency of CD73 in BGC-823 cells was confirmed by western blotting analysis. ( F ) Effect of CD73 overexpression on the glucose uptake (n = 3), lactate release (n = 3), and expression of glycolytic genes (n = 3) in BGC-823 cells. ( G ) The effect of APCP (50 μM) and adenosine (10 μM) on the glucose uptake of HGC-27 and SGC-7901 cells. ( H ) The effect of APCP (50 μM) and adenosine (10 μM) on the lactate release of HGC-27 and SGC-7901 cells. *p < 0.05; **p < 0.01.

Article Snippet: APCP (CD73 inhibitor) and adenosine were acquired from Sigma (Shanghai, China).

Techniques: Western Blot, Expressing, Over Expression

Journal: Journal of Cancer

Article Title: CD73 is a hypoxia-responsive gene and promotes the Warburg effect of human gastric cancer cells dependent on its enzyme activity

doi: 10.7150/jca.62387

Figure Lengend Snippet: The growth advantage induced by CD73 is glycolysis-dependent in gastric cancer. ( A ) The effect of CD73 knockdown on in vitro proliferation of HGC-27 and SGC-7901 cells was analyzed by colony formation assay (n = 3). ( B ) The effect of CD73 overexpression on the proliferation of BGC-823 cells in the presence or absence of 50 mM 2-DG was analyzed by plate colony formation assay. ( C ) BGC-823 cells were cultured in media containing 25 mM galactose but no glucose and plate colony formation experiment was performed. ( D ) The effect of 2-DG on the proliferation of HGC27 cells in the presence or absence of 50μM APCP was analyzed by plate colony formation assay. **p < 0.01; ***p < 0.001.

Article Snippet: APCP (CD73 inhibitor) and adenosine were acquired from Sigma (Shanghai, China).

Techniques: In Vitro, Colony Assay, Over Expression, Cell Culture

Journal: Journal of Cancer

Article Title: CD73 is a hypoxia-responsive gene and promotes the Warburg effect of human gastric cancer cells dependent on its enzyme activity

doi: 10.7150/jca.62387

Figure Lengend Snippet: CD73 knockdown suppresses tumor growth in vivo. ( A ) A subcatenous xenograft model showed the effect of CD73 knockdown on the in vivo tumor growth of HGC-27 cells (n=5). ( B ) Measurement of tumor weight and tumor volume in sh-Ctrl, sh-CD73 #1 and sh-CD73 #2 groups. ( C ) IHC analysis of Ki67 in sh-Ctrl, sh-CD73 #1 and sh-CD73 #2 tumor tissues. ( D ) Real-time qPCR analysis of glycolytic genes (GLUT1, HK2, ENO1, PKM2, and LDHA) in sh-Ctrl, sh-CD73 #1 and sh-CD73 #2 tumor tissues; GLUT1, glucose transporter type 1; HK2, hexokinase 2; ENO1, enolase 1; PKM2, pyruvate kinase M2; LDHA, lactate dehydrogenase A. *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: APCP (CD73 inhibitor) and adenosine were acquired from Sigma (Shanghai, China).

Techniques: In Vivo

Journal: Purinergic Signalling

Article Title: Hydrolysis of ATP, ADP, and AMP is increased in blood plasma of prostate cancer patients

doi: 10.1007/s11302-018-9642-3

Figure Lengend Snippet: Effect of specific inhibitors of NTPDases, CD73, and alkaline phosphatase on nucleotide hydrolysis. a Incubation of ATP plus ARL67153 (83 μM) and LEV (1 mM). b Incubation of AMP plus APCP (30 μM) and LEV (1 mM). The nucleotide hydrolysis was evaluated in the blood plasma as described in the “Methods” section, and final values were represented as nmol Pi/min/mg protein. The experiments were performed in duplicate with n = 5. Data were expressed as mean ± SEM and analyzed by one-way ANOVA, followed by Tukey’s post-hoc test. * represents the significant difference in relation to AMP hydrolysis (* p < 0.05)

Article Snippet: Chemicals Adenosine-5′triphosphate (ATP), adenosine-5′diphosphate (ADP), adenosine-5′monophosphate (AMP), α-β-methylene-ADP (APCP; inhibitor of CD73), {"type":"entrez-protein","attrs":{"text":"ARL67156","term_id":"1186396857","term_text":"ARL67156"}} ARL67156 (inhibitor of NTPDases), levamisole (inhibitor of alkaline phosphatase), Malachite Green, Coomassie Blue, and Tris–HCl were purchased from Sigma Aldrich.

Techniques: Incubation

Journal: Purinergic Signalling

Article Title: Hydrolysis of ATP, ADP, and AMP is increased in blood plasma of prostate cancer patients

doi: 10.1007/s11302-018-9642-3

Figure Lengend Snippet: The expression of CD39 and CD73 on MVs was evaluated by flow cytometry. Representative flow cytometry dot-plots showed the MVs labeled to CD39 (a) and CD73 (c). The percentage of labeling was demonstrated at (b) and (d), respectively. Annexin-V was used to identify phosphatidylserine that is present at the MV membrane [20]. Data were expressed as mean ± SEM (n = 6) and analyzed by two-way ANOVA, followed by Bonferroni’s post-hoc test

Article Snippet: Chemicals Adenosine-5′triphosphate (ATP), adenosine-5′diphosphate (ADP), adenosine-5′monophosphate (AMP), α-β-methylene-ADP (APCP; inhibitor of CD73), {"type":"entrez-protein","attrs":{"text":"ARL67156","term_id":"1186396857","term_text":"ARL67156"}} ARL67156 (inhibitor of NTPDases), levamisole (inhibitor of alkaline phosphatase), Malachite Green, Coomassie Blue, and Tris–HCl were purchased from Sigma Aldrich.

Techniques: Expressing, Flow Cytometry, Labeling