Identification of natural compounds targeting Annexin A2 with an anti-cancer effect

Abstract

Annexin A2, a multifunctional tumor associated protein, promotes nuclear factor-kappa B (NF-κB) activation by interacting with NF-κB p50 subunit and facilitating its nuclear translocation. Here we demonstrated that two ginsenosides Rg5 (G-Rg5) and Rk1 (G-Rk1), with similar structure, directly bound to Annexin A2 by molecular docking and cellular thermal shift assay. Both Rg5 and Rk1 inhibited the interaction between Annexin A2 and NF-κB p50 subunit, their translocation to nuclear and NF-κB activation. Inhibition of NF-κB by these two ginsenosides decreased the expression of inhibitor of apoptosis proteins (IAPs), leading to caspase activation and apoptosis. Over expression of K302A Annexin A2, a mutant version of Annexin A2, which fails to interact with G-Rg5 and G-Rk1, effectively reduced the NF-κB inhibitory effect and apoptosis induced by G-Rg5 and G-Rk1. In addition, the knockdown of Annexin A2 largely enhanced NF-κB activation and apoptosis induced by the two molecules, indicating that the effects of G-Rg5 and G-Rk1 on NF-κB were mainly mediated by Annexin A2. Taken together, this study for the first time demonstrated that G-Rg5 and G-Rk1 inhibit tumor cell growth by targeting Annexin A2 and NF-κB pathway, and G-Rg5 and G-Rk1 might be promising natural compounds for targeted cancer therapy.

Keywords

Annexin A2 / G-Rg5 / G-Rk1 / HCC / NF-κB

References

[1] Alexandersavino CV, Hayden MS, Richardson C (2016) Doxycycline is an NF-κB inhibitor that induces apoptotic cell death in malignant T-cells. Oncotarget 7(46):75954–75967

[2] Attele AS, Wu JA, Yuan CS (1999) Ginseng pharmacology: multiple constituents and multiple actions. Biochem Pharmacol 58:1685–1693

[3] Chen CY, Lin YS, Chen CL (2015) Targeting annexin A2 reduces tumorigenesis and therapeutic resistance of nasopharyngeal carcinoma. Oncotarget 6(29):26946–26959

[4] Chuthapisith S, Bean BE, Cowley G (2009) Annexins in human breast cancer: Possible predictors of pathological response to neoadjuvant chemotherapy. Eur J Cancer 45:1274–1281

[5] Das S, Shetty P, Valapala M (2010) Signal transducer and activator of transcription 6 (STAT6) is a novel interactor of Annexin A2 in prostate cancer cells. Biochemistry 49(10):2216–2226

[6] Deng S, Wang J, Hou L, Li J, Chen G, Jing B, Zhang X, Yang Z (2013) Annexin A1, A2, A4 and A5 play important roles in breast cancer, pancreatic cancer and laryngeal carcinoma, alone and/or synergistically. Oncol Lett 5:107–112

[7] Dolcet X, Llobet D, Pallares J, Matias-Guiu X (2005) NF-κB in development and progression of human cancer. Virchows Arch 446:475–482

[8] Duncan R, Carpenter B, Main LC, Telfer C, Murray GI (2008) Characterisation and protein expression profiling of annexins in colorectal cancer. Br J Cancer 98:426–433

[9] Ecsédi P, Kiss B, Gógl G (2017) Regulation of the equilibrium between closed and open conformations of Annexin A2 by N-terminal phosphorylation and S100A4-binding. Structure 25 (8):1195.e5–1207.e5

[10] Gerke V, Creutz CE, Moss SE (2005) Annexins: linking Ca2+ signalling to membrane dynamics. Nat Rev Mol Cell Biol 6:449–461[PubMed: 15928709]

[11] Hanahan D, Weinberg RA (2000) The hallmarks of cancer. Cell 100:57–70

[12] Hayden MS, Ghosh S (2008) Shared principles in NF-κB signaling. Cell 132(3):344–362

[13] Jung H, Kim JS, Kim WK (2015) Intracellular annexin A2 regulates NF-κB signaling by binding to the p50 subunit: implications for gemcitabine resistance in pancreatic cancer. Cell Death Dis 6(1):e1606

[14] Kesavan K, Ratliff J, Johnson EW (2010) Annexin A2 is a molecular target for TM601, a peptide with tumor-targeting and anti-angiogenic effects. J Biol Chem 285(7):4366

[15] Klionsky DJ (2012) Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy 8:445–544

[16] Kwon M, Jang H, Kim EH (2016) Efficacy of poly (ADP-ribose) polymerase inhibitor olaparib against head and neck cancer cells: Predictions of drug sensitivity based on PAR-p53-NF-κB interactions. Cell Cycle 15(22):3105

[17] Lee SM (2015) Anti-inflammatory effects of ginsenosides Rg5, Rz1, and Rk1: inhibition of TNF-α-induced NF-κB, COX-2, and iNOS transcriptional expression. Phytother Res 28(12):1893–1896

[18] Longerich T, Haller MT, Mogler C (2011) Annexin A2 as a differential diagnostic marker of hepatocellular tumors. Pathol Res Pract 207(1):8–14

[19] Ma RL, Shen LY, Chen KN (2014) Coexpression of ANXA2, SOD2 and HOXA13 predicts poor prognosis of esophageal squamous cell carcinoma. Oncol Rep 31:2157–2164

[20] Moreau K, Ravikumar B, Renna M, Puri C, Rubinsztein DC (2011) Autophagosome precursor maturation requires homotypic fusion. Cell 146:303–317

[21] Moss SE, Morgan RO (2004) The annexins. Genome Biol 5:219. [PubMed: 15059252]

[22] Ohno Y, Izumi M, Kawamura T, Nishimura T, Mukai K, Tachibana M (2009) Annexin II represents metastatic potential in clearcell renal cell carcinoma. Br J Cancer 101:287–294

[23] Perkins ND (2012) The diverse and complex roles of NF-κB subunits in cancer. Nat Rev Cancer 12:121–132

[24] Réty S, Sopkova J, Renouard M, Osterloh D, Gerke V, Tabaries S, Russo-Marie F, Lewit-Bentley A (1999) The crystal structure of a complex of p11 with the annexin II N-terminal peptide. Nat Struct Biol 6:89–95

[25] Shetty PK, Thamake SI, Biswas S, Johansson SL, Vishwanatha JK (2012) Reciprocal regulation of annexin A2 and EGFR with Her-2 in Her-2 negative and herceptin-resistant breast cancer. PLoS One 7:e44299

[26] Staquicini DI, Rangel R, Guzmanrojas L (2017) Intracellular targeting of annexin A2 inhibits tumor cell adhesion, migration, and in vivo grafting. Sci Rep 7(1):4243

[27] Sun Y, Gao G, Cai J (2013) Annexin A2 is a discriminative serological candidate in early hepatocellular carcinoma. Carcinogenesis 34(3):595–604

[28] Takano S, Togawa A, Yoshitomi H (2008) Annexin II overexpression predicts rapid recurrence after surgery in pancreatic cancer patients undergoing gemcitabine-adjuvant chemotherapy. Ann Surg Oncol 15:3157–3168

[29] Toh DF, New LS, Koh HL, Chan EC (2010) Ultra-high performance liquid chromatography/time-of-flight mass spectrometry (UHPLC/TOFMS) for time-dependent profiling of raw and steamed Panax notoginseng. J Pharm Biomed Anal 52:43–50

[30] Tong W, Jie Y, Jie Z (2015) Anxa2 binds to STAT3 and promotes epithelial to mesenchymal transition in breast cancer cells. Oncotarget 6(31):30975–30992

[31] Wang CY, Chen CL, Tseng YL (2012) Annexin A2 silencing induces G2 arrest of non-small cell lung cancer cells through p53-dependent and-independent mechanisms. J Biol Chem 287 (39):32512–32524

[32] Woo SM, Min KJ, Seo BR, Kwon TK (2016) Ym155 sensitizes trailinduced apoptosis through cathepsin s-dependent down-regulation of mcl-1 and n NF-κB down-regulation of c-flip expression in human renal carcinoma caki cells. Oncotarget 7(38):61520–61532

[33] Yang T, Peng H, Wang J, Yang J, Nice EC, Xie K, Huang C(2013) Prognostic and diagnostic significance of annexin A2 in colorectal cancer. Colorectal Dis 15:e373–e381

[34] Yang SF, Hsu HL, Chao TK, Hsiao CJ, Lin YF, Cheng CW (2014) Annexin A2 in renal cell carcinoma: Expression, function, and prognostic significance. Urol Oncol 33:22.e11–22.e21

[35] Yao H, Zhang Z, Xiao Z (2009) Identification of metastasis associated proteins in human lung squamous carcinoma using two-dimensional difference gel electrophoresis and laser capture microdissection. Lung Cancer 65:41–48

[36] Yin DL, Liang YJ, Zheng TS (2016) EF24 inhibits tumor growth and metastasis via suppressing NF-kappaB dependent pathways in human cholangiocarcinoma. Sci Rep 6:32167

[37] Zhang HJ, Yao DF, Yao M, Huang H, Wu W, Yan MJ, Yan XD, Chen J (2012a) Expression characteristics and diagnostic value of annexin A2 in hepatocellular carcinoma. World J Gastroenterol 18:5897–5904

[38] Zhang Q, Ye Z, Yang Q, He X, Wang H, Zhao Z (2012b) Upregulated expression of annexin II is a prognostic marker for patients with gastric cancer. World J Surg Oncol 10:103

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