GDF11/BMP11 activates both smad1/5/8 and smad2/3 signals but shows no significant effect on proliferation and migration of human umbilical vein endothelial cells

Abstract

GDF11/BMP11, a member of TGF-β superfamily, was reported to rejuvenate heart, skeletal muscle and blood vessel architecture in aged mice. However, the rejuvenative effects of GDF11 were questioned recently. Here, we investigated the effects of GDF11 on smad and non-smad signals in human umbilical vein endothelial cells (HUVECs) and the effects of GDF11 on proliferation and migration of HUVECs and primary rat aortic endothelial cells (RAECs). GDF11 factor purchased from two different companies (PeproTech and R&D Systems) was comparatively studied. Western blot was used to detect the protein expressions. The cell viability and migration were examined by using MTT and wound healing assays. Results showed that GDF11 activated both smad1/5/8 and smad2/3 signals in HUVECs. GDF11 increased protein expression of NADPH oxidase 4(NOX4) in HUVECs. GDF11 showed no significant effect on the protein level of p38, p-p38, ERK, p-ERK, Akt, p-Akt (Ser473) and p-Akt(Thr308), but increased the protein level of p-JNK and p-AMPK in HUVECs, and these increases were inhibited by antioxidant mitoTEMPO treatment. GDF11 slightly increased cell viability after short-term treatment and slightly decreased cell viability after long-term treatment. GDF11 showed no significant effect on cell proliferation and migration. These data indicated that the notion of GDF11 as a rejuvenation-related factor for endothelial cells needs to be cautious.

GDF11 Protects against Endothelial Injury and Reduces Atherosclerotic Lesion Formation in Apolipoprotein E-Null Mice

“…Recently, another study showed that GDF11 had no effect on proliferation and migration of human umbilical vein endothelial cell and rat aortic endothelial cells after 72 hours of treatment. 28 We also found GDF11 had no effect on the migration of MAECs and macrophages, however, we found GDF11 significantly promoted MAECs proliferation after 5 days of treatment, which makes endothelial cells have a certain ability to self-repair. The discrepancy may be due to different treatment time.…”

GDF11/BMP11 as a novel tumor marker for liver cancer

“…Several HDAC inhibitors, including vorinostat (SAHA), romidepsin, belinostat and panobinostat, have been approved by the USA food and drug administration to treat cancer (29). The mechanisms of action of HDAC inhibitors may include inhibition of abnormal cell growth by inactivation of HDAC3 and activation of GDF11 expression (14). The function of GDF11 in various cancer types remains controversial.…”

“…Recombinant GDF11 pure protein used for treating the cells was purchased from PeproTech Inc. and for the preparation of a stock solution, 20 µg GDF11 was dissolved in 40 µl sterile ultra-pure water according to the manufacturer's protocol. Recombinant GDF11 pure protein was utilized according to methods described in previous studies (13,14). Cells in the control group were treated with the same volume of sterile ultra-pure water.…”

“…Western blot analysis was performed as described in previous studies by our group (14,22). Cells were lysed with radioimmunoprecipitation assay buffer (Beyotime Institute of Biotechnology, Haimen, China) and centrifuged at 12,000 x g for 15 min at 4˚C.…”

“…The Smad protein complex then translocates into the nucleus to positively or negatively control gene expression (9,10). GDF11 not only contributes to embryonic development and histogenesis, but also has a role in metabolic disorders, cardiovascular diseases and ageing (11)(12)(13)(14). GDF11 is expressed in the liver and has an inhibitory role in liver development.…”