Different roles of Akt and mechanistic target of rapamycin in serum‑dependent chondroprotection of human osteoarthritic chondrocytes

Zhang,Tongen; Liu,Jijie; Zheng,Xinpeng; Zhang,Bing; Xia,Chun

doi:10.3892/ijmm.2017.3285

Different roles of Akt and mechanistic target of rapamycin in serum‑dependent chondroprotection of human osteoarthritic chondrocytes

Authors:
- Tongen Zhang
- Jijie Liu
- Xinpeng Zheng
- Bing Zhang
- Chun Xia
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Affiliations: Department of Sports Medicine and Joint Surgery, Zhongshan Hospital, Xiamen University, Xiamen, Fujian 361004, P.R. China, Medical School, Xiamen University, Xiamen, Fujian 361102, P.R. China
Published online on: November 24, 2017 https://doi.org/10.3892/ijmm.2017.3285
Pages: 977-984

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Abstract

Despite various animal serums being used widely to culture chondrocytes, the regulatory mechanism of serum on chondrocyte activities has not been elucidated. In the present study, human osteoarthritis (OA) chondrocytes were used to perform in vitro investigations on the effect of different concentrations of bovine fetal serum on extracellular matrix synthesis, cell proliferation and autophagy using the Cell Counting Kit‑8 analysis, a laser‑scanning confocal microscope, and western blot analysis. The results demonstrated that 5% serum exerted a chondroprotective effect more than the other concentrations of serum, as it simultaneously promoted cell proliferation, autophagy, and ECM synthesis in human OA chondrocytes. Furthermore, the decreased mechanistic target of rapamycin (mTOR) and increased Akt were observed in 5% serum‑treated OA chondrocytes. Either mTOR or Akt inhibitor influenced the effect of 5% serum on cell proliferation and autophagy in human OA chondrocytes, which was associated with LC‑3B or B‑cell lymphoma-2 (Bcl‑2) signal molecules. Consistent with previous studies, the present study proposes that 5% serum promotes cell proliferation via the Akt/Bcl‑2 axis and induces autophagy via the mTOR/LC‑3B axis in human OA chondrocytes. Furthermore, the different roles of Akt and mTOR in the cell processes of human OA chondrocytes require consideration for preclinical and clinical therapy of OA.

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1	Levine B and Kroemer G: Autophagy in the pathogenesis of disease. Cell. 132:27–42. 2008. View Article : Google Scholar : PubMed/NCBI
2	Cinque L, Forrester A, Bartolomeo R, Svelto M, Venditti R, Montefusco S, Polishchuk E, Nusco E, Rossi A, Medina DL, et al: FGF signalling regulates bone growth through autophagy. Nature. 528:272–275. 2015. View Article : Google Scholar : PubMed/NCBI
3	Sasaki H, Takayama K, Matsushita T, Ishida K, Kubo S, Matsumoto T, Fujita N, Oka S, Kurosaka M and Kuroda R: Autophagy modulates osteoarthritis-related gene expression in human chondrocytes. Arthritis Rheum. 64:1920–1928. 2012. View Article : Google Scholar
4	Zhao H, Zhang T, Xia C, Shi L, Wang S, Zheng X, Hu T and Zhang B: Berberine ameliorates cartilage degeneration in interleukin-1β-stimulated rat chondrocytes and in a rat model of osteoarthritis via Akt signalling. J Cell Mol Med. 18:283–292. 2014. View Article : Google Scholar
5	Cheng L, Zeng G, Liu Z, Zhang B, Cui X, Zhao H, Zheng X, Song G, Kang J and Xia C: Protein kinase B and extracellular signal-regulated kinase contribute to the chondroprotective effect of morroniside on osteoarthritis chondrocytes. J Cell Mol Med. 19:1877–1886. 2015. View Article : Google Scholar : PubMed/NCBI
6	Zhou PH, Liu SQ and Peng H: The effect of hyaluronic acid on IL-1beta-induced chondrocyte apoptosis in a rat model of osteoarthritis. J Orthop Res. 26:1643–1648. 2008. View Article : Google Scholar : PubMed/NCBI
7	Ribeiro M, López de Figueroa P, Nogueira-Recalde U, Centeno A, Mendes AF, Blanco FJ and Caramés B: Diabetes-accelerated experimental osteoarthritis is prevented by autophagy activation. Osteoarthritis Cartilage. 24:2116–2125. 2016. View Article : Google Scholar : PubMed/NCBI
8	Zhang Y, Vasheghani F, Li YH, Blati M, Simeone K, Fahmi H, Lussier B, Roughley P, Lagares D, Pelletier JP, et al: Cartilage-specific deletion of mTOR upregulates autophagy and protects mice from osteoarthritis. Ann Rheum Dis. 74:1432–1440. 2015. View Article : Google Scholar
9	Cui X, Wang S, Cai H, Lin Y, Zheng X, Zhang B and Xia C: Overexpression of microRNA-634 suppresses survival and matrix synthesis of human osteoarthritis chondrocytes by targeting PIK3R1. Sci Rep. 6:231172016. View Article : Google Scholar : PubMed/NCBI
10	Chen J, Crawford R and Xiao Y: Vertical inhibition of the PI3K/Akt/mTOR pathway for the treatment of osteoarthritis. J Cell Biochem. 114:245–249. 2013. View Article : Google Scholar
11	Baltzer AW, Moser C, Jansen SA and Krauspe R: Autologous conditioned serum (Orthokine) is an effective treatment for knee osteoarthritis. Osteoarthritis Cartilage. 17:152–160. 2009. View Article : Google Scholar
12	Moser C: Response to: cytokine profile of autologous conditioned serum for treatment of osteoarthritis, in vitro effects on cartilage metabolism and intra-articular levels after injection. Arthritis Res Ther. 12:4102010. View Article : Google Scholar
13	Rutgers M, Saris DB, Dhert WJ and Creemers LB: Cytokine profile of autologous conditioned serum for treatment of osteoarthritis, in vitro effects on cartilage metabolism and intra-articular levels after injection. Arthritis Res Ther. 12:R1142010. View Article : Google Scholar : PubMed/NCBI
14	Attur MG, Dave M, Cipolletta C, Kang P, Goldring MB, Patel IR, Abramson SB and Amin AR: Reversal of autocrine and paracrine effects of interleukin 1 (IL-1) in human arthritis by type II IL-1 decoy receptor. Potential for pharmacological intervention. J Biol Chem. 275:40307–40315. 2000. View Article : Google Scholar : PubMed/NCBI
15	Zeng G, Cui X, Liu Z, Zhao H, Zheng X, Zhang B and Xia C: Disruption of phosphoinositide-specific phospholipases Cγ1 contributes to extracellular matrix synthesis of human osteoarthritis chondrocytes. Int J Mol Sci. 15:13236–13246. 2014. View Article : Google Scholar : PubMed/NCBI
16	Li C, Cai S, Wang X and Jiang Z: Identification and characterization of ANO9 in stage II and III colorectal carcinoma. Oncotarget. 6:29324–29334. 2015. View Article : Google Scholar : PubMed/NCBI
17	Zheng CH and Levenston ME: Fact versus artifact: Avoiding erroneous estimates of sulfated glycosaminoglycan content using the dimethylmethylene blue colorimetric assay for tissue-engineered constructs. Eur Cell Mater. 29:224–236. 2015. View Article : Google Scholar : PubMed/NCBI
18	Zhang B and Xia C: 12-O-tetradecanoylphorbol-1, 3-acetate induces the negative regulation of protein kinase B by protein kinase Calpha during gastric cancer cell apoptosis. Cell Mol Biol Lett. 15:377–394. 2010. View Article : Google Scholar : PubMed/NCBI
19	Shao XX, Duncan NA, Lin L, Fu X, Zhang JY and Yu CL: Serum-free media for articular chondrocytes in vitro expansion. Chin Med J (Engl). 126:2523–2529. 2013.
20	Wolf F, Haug M, Farhadi J, Candrian C, Martin I and Barbero A: A low percentage of autologous serum can replace bovine serum to engineer human nasal cartilage. Eur Cell Mater. 15:1–10. 2008. View Article : Google Scholar : PubMed/NCBI
21	Jain MV, Paczulla AM, Klonisch T, Dimgba FN, Rao SB, Roberg K, Schweizer F, Lengerke C, Davoodpour P, Palicharla VR, et al: Interconnections between apoptotic, autophagic and necrotic pathways: Implications for cancer therapy development. J Cell Mol Med. 17:12–29. 2013. View Article : Google Scholar : PubMed/NCBI
22	Laplante M and Sabatini DM: mTOR signaling in growth control and disease. Cell. 149:274–293. 2012. View Article : Google Scholar : PubMed/NCBI
23	Inoki K, Li Y, Xu T and Guan KL: Rheb GTPase is a direct target of TSC2 GAP activity and regulates mTOR signaling. Genes Dev. 17:1829–1834. 2003. View Article : Google Scholar : PubMed/NCBI
24	Tee AR, Manning BD, Roux PP, Cantley LC and Blenis J: Tuberous sclerosis complex gene products, Tuberin and Hamartin, control mTOR signaling by acting as a GTPase-activating protein complex toward Rheb. Curr Biol. 13:1259–1268. 2003. View Article : Google Scholar : PubMed/NCBI
25	Bohensky J, Leshinsky S, Srinivas V and Shapiro IM: Chondrocyte autophagy is stimulated by HIF-1 dependent AMPK activation and mTOR suppression. Pediatr Nephrol. 25:633–642. 2010. View Article : Google Scholar :
26	Xue JF, Shi ZM, Zou J and Li XL: Inhibition of PI3K/AKT/mTOR signaling pathway promotes autophagy of articular chondrocytes and attenuates inflammatory response in rats with osteoarthritis. Biomed Pharmacother. 89:1252–1261. 2017. View Article : Google Scholar : PubMed/NCBI
27	Takayama K, Kawakami Y, Kobayashi M, Greco N, Cummins JH, Matsushita T, Kuroda R, Kurosaka M, Fu FH and Huard J: Local intra-articular injection of rapamycin delays articular cartilage degeneration in a murine model of osteoarthritis. Arthritis Res Ther. 16:4822014. View Article : Google Scholar : PubMed/NCBI
28	Rosa SC, Rufino AT, Judas F, Tenreiro C, Lopes MC and Mendes AF: Expression and function of the insulin receptor in normal and osteoarthritic human chondrocytes: Modulation of anabolic gene expression, glucose transport and GLUT-1 content by insulin. Osteoarthritis Cartilage. 19:719–727. 2011. View Article : Google Scholar : PubMed/NCBI
29	Cravero JD, Carlson CS, Im HJ, Yammani RR, Long D and Loeser RF: Increased expression of the Akt/PKB inhibitor TRB3 in osteoarthritic chondrocytes inhibits insulin-like growth factor 1-mediated cell survival and proteoglycan synthesis. Arthritis Rheum. 60:492–500. 2009. View Article : Google Scholar : PubMed/NCBI
30	Hahn-Windgassen A, Nogueira V, Chen CC, Skeen JE, Sonenberg N and Hay N: Akt activates the mammalian target of rapamycin by regulating cellular ATP level and AMPK activity. J Biol Chem. 280:32081–32089. 2005. View Article : Google Scholar : PubMed/NCBI
31	Peng M, Huang Y, Tao T, Peng CY, Su Q, Xu W, Darko KO, Tao X and Yang X: Metformin and gefitinib cooperate to inhibit bladder cancer growth via both AMPK and EGFR pathways joining at Akt and Erk. Sci Rep. 6:286112016. View Article : Google Scholar : PubMed/NCBI
32	Galasso O, Panza S, Santoro M, Goldring MB, Aquila S and Gasparini G: PTEN elevation, autophagy and metabolisc reprogramming may be induced in human chondrocytes during steroids or nutrient depletion and osteoarthritis. J Biol Regul Homeost Agents. 29(Suppl 4): S1–S14. 2015.
33	Memmott RM and Dennis PA: Akt-dependent and -independent mechanisms of mTOR regulation in cancer. Cell Signal. 21:656–664. 2009. View Article : Google Scholar : PubMed/NCBI
34	O'Reilly KE, Rojo F, She QB, Solit D, Mills GB, Smith D, Lane H, Hofmann F, Hicklin DJ, Ludwig DL, et al: mTOR inhibition induces upstream receptor tyrosine kinase signaling and activates Akt. Cancer Res. 66:1500–1508. 2006. View Article : Google Scholar : PubMed/NCBI
35	Yori JL, Lozada KL, Seachrist DD, Mosley JD, Abdul-Karim FW, Booth CN, Flask CA and Keri RA: Combined SFK/mTOR inhibition prevents rapamycin-induced feedback activation of AKT and elicits efficient tumor regression. Cancer Res. 74:4762–4771. 2014. View Article : Google Scholar : PubMed/NCBI