OBJECTIVE: To determine whether erythropoietin (EPO) promotes rapid proliferation of glioma through Akt pathway. METHODS: We detected the expression of EPO in human glioma tissues using immunohistochemistry. A nude mouse model bearing human glioma U87 cell xenograft was established and given intraperitoneal injection of EPO or saline every other day, and the tumor growth was observed. In the in vitro experiment, U87 cells were treated with PBS (control), EPO, or EPO with Akt inhibitor, and the expression of p-Akt and cyclin D1 was detected using Western blotting; the cell proliferation rate was determined using cell counting kit-8 and clone formation assay, and the cell cycle changes were analyzed with flow cytometry. RESULTS: Compared with low-grade glioma tissues, high-grade glioma tissues exhibited a significantly increased EPO expression (P=0.0002). In the tumor-bearing mice, EPO treatment significantly increased the expression of EPO (P=0.0006) and p-Akt (P=0.0003) in the tumor and obviously increased the tumor volume (P<0.0001) and weight (P=0.0003). In U87 cells cultured in vitro, EPO treatment obviously accelerated the cell proliferation (P=0.020 on day 3 and 0.028 on day 5), promoted clone formation (P=0.0010), and increased proliferation index (P=0.0028); EPO significantly enhanced the protein expression of p-Akt (P=0.0020) and cyclin D1 (P=0.0022). The application of Akt inhibitor significantly suppressed the effect of EPO in enhancing cyclin D1 and p-Akt expression (both P<0.0001) and promoting cell proliferation. CONCLUSION: EPO can significantly accelerate the proliferation of glioma through Akt pathway.
OBJECTIVE: To determine whether erythropoietin (EPO) promotes rapid proliferation of glioma through Akt pathway. METHODS: We detected the expression of EPO in humanglioma tissues using immunohistochemistry. A nude mouse model bearing humanglioma U87 cell xenograft was established and given intraperitoneal injection of EPO or saline every other day, and the tumor growth was observed. In the in vitro experiment, U87 cells were treated with PBS (control), EPO, or EPO with Akt inhibitor, and the expression of p-Akt and cyclin D1 was detected using Western blotting; the cell proliferation rate was determined using cell counting kit-8 and clone formation assay, and the cell cycle changes were analyzed with flow cytometry. RESULTS: Compared with low-grade glioma tissues, high-grade glioma tissues exhibited a significantly increased EPO expression (P=0.0002). In the tumor-bearing mice, EPO treatment significantly increased the expression of EPO (P=0.0006) and p-Akt (P=0.0003) in the tumor and obviously increased the tumor volume (P<0.0001) and weight (P=0.0003). In U87 cells cultured in vitro, EPO treatment obviously accelerated the cell proliferation (P=0.020 on day 3 and 0.028 on day 5), promoted clone formation (P=0.0010), and increased proliferation index (P=0.0028); EPO significantly enhanced the protein expression of p-Akt (P=0.0020) and cyclin D1 (P=0.0022). The application of Akt inhibitor significantly suppressed the effect of EPO in enhancing cyclin D1 and p-Akt expression (both P<0.0001) and promoting cell proliferation. CONCLUSION:EPO can significantly accelerate the proliferation of glioma through Akt pathway.
Authors: Sunila Pradeep; Jie Huang; Edna M Mora; Alpa M Nick; Min Soon Cho; Sherry Y Wu; Kyunghee Noh; Chad V Pecot; Rajesha Rupaimoole; Martin A Stein; Stephan Brock; Yunfei Wen; Chiyi Xiong; Kshipra Gharpure; Jean M Hansen; Archana S Nagaraja; Rebecca A Previs; Pablo Vivas-Mejia; Hee Dong Han; Wei Hu; Lingegowda S Mangala; Behrouz Zand; Loren J Stagg; John E Ladbury; Bulent Ozpolat; S Neslihan Alpay; Masato Nishimura; Rebecca L Stone; Koji Matsuo; Guillermo N Armaiz-Peña; Heather J Dalton; Christopher Danes; Blake Goodman; Cristian Rodriguez-Aguayo; Carola Kruger; Armin Schneider; Shyon Haghpeykar; Padmavathi Jaladurgam; Mien-Chie Hung; Robert L Coleman; Jinsong Liu; Chun Li; Diana Urbauer; Gabriel Lopez-Berestein; David B Jackson; Anil K Sood Journal: Cancer Cell Date: 2015-10-17 Impact factor: 31.743