Blockade of phospholipid scramblase 1 with its N-terminal domain antibody reduces tumorigenesis of colorectal carcinomas in vitro and in vivo

Journal of Translational Medicine

Err-Cheng Chan²  Wei-Shan Wei³  Yeh-Pin Chou⁴  Ya-Shan Chen¹  Yung-Bin Kuo²  Chia-Rui Shen²  Kuei-Tien Chen²  Chun-Yu Chen¹  Chung-Wei Fan⁵ 

[1] Graduate Institute of Biomedical Sciences, Chang Gung University, Taoyuan, Taiwan;Department of Medical Biotechnology and Laboratory Science, Chang Gung University, Taoyuan, Taiwan;Graduate Institute of Biochemical and Biomedical Engineering, Chang Gung University, Taoyuan, Taiwan;Division of Hepato-Gastroenterology, Chang Gung Memorial Hospital, Kaohsiung Medical Center, Kaohsiung, Taiwan;College of Medicine, Chang Gung University, Taoyuan, Taiwan

关键词: Colorectal carcinomas;    Cell cycle G1/S arrest;    Cyclin D1;    Retinoblastoma dephosphorylation;    Phospholipid scramblase 1;   

Others  :  828283
DOI  :  10.1186/1479-5876-10-254

 received in 2012-10-21, accepted in 2012-12-19,  发布年份 2012

【 摘 要 】

Background

Membrane-bound phospholipid scramblase 1 (PLSCR1) is involved in both lipid trafficking and cell signaling. Previously, we showed that PLSCR1 is overexpressed in many colorectal carcinomas (CRCs). In the present study, we investigated the tumorigenic role of PLSCR1 in CRC and suggest that it is a potential therapeutic target.

Methods

To identify PLSCR1 as a therapeutic target, we studied the tumorigenic properties of CRC cell lines treated with a monoclonal antibody (NP1) against the N-terminus of PLSCR1 in vitro and in vivo. We also investigated cell cycle status and epidermal growth factor receptor–related pathways and downstream effectors of PLSCR1 after blocking its function with NP1.

Results

Treating CRC cells with NP1 in vitro and in vivo decreased cell proliferation, anchorage-independent growth, migration, and invasion. Adding NP1 to the CRC cell line HT29 caused arrest at G1/S. Treating HT29 cells with NP1 significantly decreased the expression of cyclin D1 and phosphorylation levels of Src, the adaptor protein Shc, and Erks. The reduced level of cyclin D1 led to an increase in the activated form of the tumor suppressor retinoblastoma protein via dephosphorylation. These actions led to attenuation of tumorigenesis.

Conclusions

Therefore, PLSCR1 may serve as a potential therapeutic target for CRC.

【 授权许可】

   
2012 Fan et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Han CL, Chen JS, Chan EC, Wu CP, Yu KH, Chen KT, Tsou CC, Tsai CF, Chien CW, Kuo YB, Lin PY, Yu JS, Hsueh C, Chen MC, Chan CC, Chang YS, Chen YJ: An informatics-assisted label-free approach for personalized tissue membrane proteomics: case study on colorectal cancer. Mol Cell Proteomics 2011, 10:M110 003087.
• [2]Kuo YB, Chan CC, Chang CA, Fan CW, Hung RP, Hung YS, Chen KT, Yu JS, Chang YS, Chan EC: Identification of phospholipid scramblase 1 as a biomarker and determination of its prognostic value for colorectal cancer. Mol Med 2011, 17:41-47.
• [3]Basse F, Stout JG, Sims PJ, Wiedmer T: Isolation of an erythrocyte membrane protein that mediates Ca2+−dependent transbilayer movement of phospholipid. J Biol Chem 1996, 271:17205-17210.
• [4]Zhao J, Zhou Q, Wiedmer T, Sims PJ: Level of expression of phospholipid scramblase regulates induced movement of phosphatidylserine to the cell surface. J Biol Chem 1998, 273:6603-6606.
• [5]Li Y, Rogulski K, Zhou Q, Sims PJ, Prochownik EV: The negative c-Myc target onzin affects proliferation and apoptosis via its obligate interaction with phospholipid scramblase 1. Mol Cell Biol 2006, 26:3401-3413.
• [6]Zhou Q, Ben-Efraim I, Bigcas JL, Junqueira D, Wiedmer T, Sims PJ: Phospholipid scramblase 1 binds to the promoter region of the inositol 1,4,5-triphosphate receptor type 1 gene to enhance its expression. J Biol Chem 2005, 280:35062-35068.
• [7]Zhou Q, Zhao J, Wiedmer T, Sims PJ: Normal hemostasis but defective hematopoietic response to growth factors in mice deficient in phospholipid scramblase 1. Blood 2002, 99:4030-4038.
• [8]Silverman RH, Halloum A, Zhou A, Dong B, Al-Zoghaibi F, Kushner D, Zhou Q, Zhao J, Wiedmer T, Sims PJ: Suppression of ovarian carcinoma cell growth in vivo by the interferon-inducible plasma membrane protein, phospholipid scramblase 1. Cancer Res 2002, 62:397-402.
• [9]Frasch SC, Henson PM, Kailey JM, Richter DA, Janes MS, Fadok VA, Bratton DL: Regulation of phospholipid scramblase activity during apoptosis and cell activation by protein kinase Cdelta. J Biol Chem 2000, 275:23065-23073.
• [10]Pastorelli C, Veiga J, Charles N, Voignier E, Moussu H, Monteiro RC, Benhamou M: IgE receptor type I-dependent tyrosine phosphorylation of phospholipid scramblase. J Biol Chem 2001, 276:20407-20412.
• [11]Sun J, Zhao J, Schwartz MA, Wang JY, Wiedmer T, Sims PJ: c-Abl tyrosine kinase binds and phosphorylates phospholipid scramblase 1. J Biol Chem 2001, 276:28984-28990.
• [12]Nanjundan M, Sun J, Zhao J, Zhou Q, Sims PJ, Wiedmer T: Plasma membrane phospholipid scramblase 1 promotes EGF-dependent activation of c-Src through the epidermal growth factor receptor. J Biol Chem 2003, 278:37413-37418.
• [13]Yu JS, Chen WJ, Ni MH, Chan WH, Yang SD: Identification of the regulatory autophosphorylation site of autophosphorylation-dependent protein kinase (auto-kinase). Evidence that auto-kinase belongs to a member of the p21-activated kinase family. Biochem J 1998, 334:121-131.
• [14]Ravn V, Rasmussen BB, Hojholt L, Barfoed M, Heiberg I, Thorpe SM: Reproducibility of subjective immunohistochemical estrogen- and progesterone receptor determination in human endometrium. Pathol Res Pract 1993, 189:1015-1022.
• [15]Lamberti C, Lin KM, Yamamoto Y, Verma U, Verma IM, Byers S, Gaynor RB: Regulation of beta-catenin function by the IkappaB kinases. J Biol Chem 2001, 276:42276-42286.
• [16]Wiedmer T, Zhou Q, Kwoh DY, Sims PJ: Identification of three new members of the phospholipid scramblase gene family. Biochim Biophys Acta 2000, 1467:244-253.
• [17]Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D: Global cancer statistics. CA Cancer J Clin 2011, 61:69-90.
• [18]Starling N, Cunningham D: Monoclonal antibodies against vascular endothelial growth factor and epidermal growth factor receptor in advanced colorectal cancers: present and future directions. Curr Opin Oncol 2004, 16:385-390.
• [19]Shaw PH, Adams RA: Where now for anti-EGF receptor therapies in colorectal cancer? Expert Rev Anticancer Ther 2011, 11:1543-1553.
• [20]Sheng H, Shao J, Townsend CM Jr, Evers BM: Phosphatidylinositol 3-kinase mediates proliferative signals in intestinal epithelial cells. Gut 2003, 52:1472-1478.
• [21]Cui W, Li SY, Du JF, Zhu ZM, An P: Silencing phospholipid scramblase 1 expression by RNA interference in colorectal cancer and metastatic liver cancer. Hepatobiliary Pancreat Dis Int 2012, 15:393-400.
• [22]Sun J, Nanjundan M, Pike LJ, Wiedmer T, Sims PJ: Plasma membrane phospholipid scramblase 1 is enriched in lipid rafts and interacts with the epidermal growth factor receptor. Biochemistry 2002, 41:6338-6345.
• [23]Nicholson RI, Gee JM, Harper ME: EGFR and cancer prognosis. Eur J Cancer 2001, 37(Suppl 4):S9-15.
• [24]Bateman A, Finn RD, Sims PJ, Wiedmer T, Biegert A, Soding J: Phospholipid scramblases and Tubby-like proteins belong to a new superfamily of membrane tethered transcription factors. Bioinformatics 2009, 25:159-162.
• [25]Viloria-Petit A, Crombet T, Jothy S, Hicklin D, Bohlen P, Schlaeppi JM, Rak J, Kerbel RS: Acquired resistance to the antitumor effect of epidermal growth factor receptor-blocking antibodies in vivo: a role for altered tumor angiogenesis. Cancer Res 2001, 61:5090-5101.
• [26]Chakravarti A, Loeffler JS, Dyson NJ: Insulin-like growth factor receptor I mediates resistance to anti-epidermal growth factor receptor therapy in primary human glioblastoma cells through continued activation of phosphoinositide 3-kinase signaling. Cancer Res 2002, 62:200-207.
• [27]Ciardiello F, Damiano V, Bianco R, Bianco C, Fontanini G, De Laurentiis M, De Placido S, Mendelsohn J, Bianco AR, Tortora G: Antitumor activity of combined blockade of epidermal growth factor receptor and protein kinase A. J Natl Cancer Inst 1996, 88:1770-1776.
• [28]Ciardiello F, Caputo R, Bianco R, Damiano V, Pomatico G, Pepe S, Bianco AR, Agrawal S, Mendelsohn J, Tortora G: Cooperative inhibition of renal cancer growth by anti-epidermal growth factor receptor antibody and protein kinase A antisense oligonucleotide. J Natl Cancer Inst 1998, 90:1087-1094.
• [29]Ciardiello F, Bianco R, Damiano V, Fontanini G, Caputo R, Pomatico G, De Placido S, Bianco AR, Mendelsohn J, Tortora G: Antiangiogenic and antitumor activity of anti-epidermal growth factor receptor C225 monoclonal antibody in combination with vascular endothelial growth factor antisense oligonucleotide in human GEO colon cancer cells. Clin Cancer Res 2000, 6:3739-3747.
• [30]Moulder SL, Yakes FM, Muthuswamy SK, Bianco R, Simpson JF, Arteaga CL: Epidermal growth factor receptor (HER1) tyrosine kinase inhibitor ZD1839 (Iressa) inhibits HER2/neu (erbB2)-overexpressing breast cancer cells in vitro and in vivo. Cancer Res 2001, 61:8887-8895.
• [31]Normanno N, Campiglio M, De LA, Somenzi G, Maiello M, Ciardiello F, Gianni L, Salomon DS, Menard S: Cooperative inhibitory effect of ZD1839 (Iressa) in combination with trastuzumab (Herceptin) on human breast cancer cell growth. Ann Oncol 2002, 13:65-72.
• [32]Takashi K, Junko OK, Yoshio H: TRA1, a Novel mRNA Highly Expressed in Leukemogenic Mouse Monocytic Sublines But Not in Nonleukemogenic Sublines. Blood 1997, 89:2975-2985.
• [33]Tsuyoshi N, Junko OK, Yuri YY, Ken-ichiro H, Shigeru T, Yoshio H, Takashi K: Role of MmTRA1b/phospholipid scramblase1 gene expression in the induction of differentiation of human myeloid leukemia cells into granulocytes. Exp Hematol 2002, 30:421-429.
• [34]Akihiro Y, Takuya Y, Eisuke S, Atsuko N, Junko OK, Tsuyoshi N, Shigeru T, Fumihiko K, Kazuo M, Yoshio H, Takashi K: MmTRA1b/phospholipid scramblase 1 gene expression is a new prognostic factor for acute myelogenous leukemia. Leukemia Res 2004, 28:149-157.
• [35]Huang Y, Zhao Q, Zhou CX, Gu ZM, Li D, Xu HZ, Sims PJ, Zhao KW, Chen GQ: Antileukemic roles of human phospholipid scramblase 1 gene, evidence from inducible PLSCR1-expressing leukemic cells. Oncogene 2006, 25:6618-6627.
• [36]Baselga J: Monoclonal antibodies directed at growth factor receptors. nn Oncol 2000, 11(Suppl 3):187-190.
• [37]Baselga J, Averbuch SD: ZD1839 ('Iressa') as an anticancer agent. Drugs 2000, 60(Suppl 1):33-40. discussion 41–32
• [38]Ranson M, Hammond LA, Ferry D, Kris M, Tullo A, Murray PI, Miller V, Averbuch S, Ochs J, Morris C, Feyereislova A, Swaisland H, Rowinsky EK: ZD1839, a selective oral epidermal growth factor receptor-tyrosine kinase inhibitor, is well tolerated and active in patients with solid, malignant tumors: results of a phase I trial. J Clin Oncol 2002, 20:2240-2250.
• [39]Hidalgo M, Siu LL, Nemunaitis J, Rizzo J, Hammond LA, Takimoto C, Eckhardt SG, Tolcher A, Britten CD, Denis L, Ferrante K, Von Hoff DD, Silberman S, Rowinsky EK: Phase I and pharmacologic study of OSI-774, an epidermal growth factor receptor tyrosine kinase inhibitor, in patients with advanced solid malignancies. J Clin Oncol 2001, 19:3267-3279.