Depletion of OLFM4 gene inhibits cell growth and increases sensitization to hydrogen peroxide and tumor necrosis factor-alpha induced-apoptosis in gastric cancer cells

Journal of Biomedical Science

Yi Huang⁶  Li-ya Wu³  Na Ang³  Xue Jiang³  Li-wen Yue⁵  Hua-an Yang⁷  Li-ming Bao¹  Hua Xiang²  Mei-hua Yang⁴  Rui-hua Liu² 

[1] Division of Human Genetics, Cincinnati Children's Hospital Medical Center, Cincinnati 45229, USA;Ministry of Education Key Laboratory of Laboratory Medical Diagnosis, Chongqing Medical University, Chongqing 400016, China;Department of Biochemistry and Molecular Biology, Chongqing Medical University, Chongqing 400037, China;Department of Neurosurgery, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China;Department of Health Care, Nanjing Maternity and Child Health Care Hospital, Nanjing 210004, China;Center for Clinical Molecular Medicine; Ministry of Education Key Laboratory of Child Development and Disorders; Key Laboratory of Pediatrics in Chongqing; Chongqing International Science and Technology Cooperation Center for Child Development and Disorders; Children's Hospital, Chongqing Medical University, Chongqing 400014, China;Department of Urologic Surgery, Yubei Pepole's Hospital, Chongqing 401120, China

关键词: Apoptosis resistance;    Cell growth;    RNA interference;    Olfactomedin 4;    Gastric cancer;   

Others  :  825334
DOI  :  10.1186/1423-0127-19-38

 received in 2011-12-03, accepted in 2012-04-03,  发布年份 2012

【 摘 要 】

Background

Human olfactomedin 4 (OLFM4) gene is a secreted glycoprotein more commonly known as the anti-apoptotic molecule GW112. OLFM4 is found to be frequently up-regulated in many types of human tumors including gastric cancer and it was believed to play significant role in the progression of gastric cancer. Although the function of OLFM4 has been indicated in many studies, recent evidence strongly suggests a cell or tissue type-dependent role of OLFM4 in cell growth and apoptosis. The aim of this study is to examine the role of gastric cancer-specific expression of OLFM4 in cell growth and apoptosis resistance.

Methods

OLFM4 expression was eliminated by RNA interference in SGC-7901 and MKN45 cells. Cell proliferation, anchorage-independent growth, cell cycle and apoptosis were characterized in vitro. Tumorigenicity was analyzed in vivo. The apoptosis and caspase-3 activation in response to hydrogen peroxide (H₂O₂) or tumor necrosis factor-alpha (TNF α) were assessed in the presence or absence of caspase inhibitor Z-VAD-fmk.

Results

The elimination of OLFM4 protein by RNA interference in SGC-7901 and MKN45 cells significantly inhibits tumorigenicity both in vitro and in vivo by induction of cell G1 arrest (all P < 0.01). OLFM4 knockdown did not trigger obvious cell apoptosis but increased H₂O₂ or TNF α-induced apoptosis and caspase-3 activity (all P < 0.01). Treatment of Z-VAD-fmk attenuated caspase-3 activity and significantly reversed the H₂O₂ or TNF α-induced apoptosis in OLFM4 knockdown cells (all P < 0.01).

Conclusion

Our study suggests that depletion of OLFM4 significantly inhibits tumorigenicity of the gastric cancer SGC-7901 and MKN45 cells. Blocking OLFM4 expression can sensitize gastric cancer cells to H₂O₂ or TNF α treatment by increasing caspase-3 dependent apoptosis. A combination strategy based on OLFM4 inhibition and anticancer drugs treatment may provide therapeutic potential in gastric cancer intervention.

【 授权许可】

   
2012 Liu et al; licensee BioMed Central Ltd.

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【 参考文献 】

• [1]Zhang J, Liu WL, Tang DC, Chen L, Wang M, Pack SD, Zhuang Z, Rodgers GP: Identification and characterization of a novel member of olfactomedin-related protein family, hGC-1, expressed during myeloid lineage development. Gene 2002, 283:83-93.
• [2]Zhang X, Huang Q, Yang Z, Li Y, Li CY: GW112, a novel antiapoptotic protein that promotes tumor growth. Cancer Res 2004, 64:2474-2481.
• [3]Kulkarni NH, Karavanich CA, Atchley WR, Anholt RR: Characterization and differential expression of a human gene family of olfactomedin-related proteins. Genet Res 2000, 76:41-50.
• [4]Shinozaki S, Nakamura T, Iimura M, Kato Y, Iizuka B, Kobayashi M, Hayashi N: Upregulation of Reg 1alpha and GW112 in the epithelium of inflamed colonic mucosa. Gut 2001, 48:623-629.
• [5]Mannick EE, Schurr JR, Zapata A, Lentz JJ, Gastanaduy M, Cote RL, Delgado A, Correa P, Correa H: Gene expression in gastric biopsies from patients infected with Helicobacter pylori. Scand J Gastroenterol 2004, 39:1192-1200.
• [6]Liu W, Yan M, Liu Y, Wang R, Li C, Deng C, Singh A, Coleman WG Jr, Rodgers GP: Olfactomedin 4 down-regulates innate immunity against Helicobacter pylori infection. Proc Natl Acad Sci USA 2010, 107:11056-11061.
• [7]Grutzmann R, Pilarsky C, Staub E, Schmitt AO, Foerder M, Specht T, Hinzmann B, Dahl E, Alldinger I, Rosenthal A, Ockert D, Saeger HD: Systematic isolation of genes differentially expressed in normal and cancerous tissue of the pancreas. Pancreatology 2003, 3:169-178.
• [8]Kobayashi D, Koshida S, Moriai R, Tsuji N, Watanabe N: Olfactomedin 4 promotes S-phase transition in proliferation of pancreatic cancer cells. Cancer Sci 2007, 98:334-340.
• [9]Koshida S, Kobayashi D, Moriai R, Tsuji N, Watanabe N: Specific overexpression of OLFM4(GW112/HGC-1) mRNA in colon, breast and lung cancer tissues detected using quantitative analysis. Cancer Sci 2007, 98:315-320.
• [10]Liu W, Zhu J, Cao L, Rodgers GP: Expression of hGC-1 is correlated with differentiation of gastric carcinoma. Histopathology 2007, 51:157-165.
• [11]Li SR, Dorudi S, Bustin SA: Identification of differentially expressed genes associated with colorectal cancer liver metastasis. Eur Surg Res 2003, 35:327-336.
• [12]Wentzensen N, Wilz B, Findeisen P, Wagner R, Dippold W, von Knebel Doeberitz M, Gebert J: Identification of differentially expressed genes in colorectal adenoma compared to normal tissue by suppression subtractive hybridization. Int J Oncol 2004, 24:987-994.
• [13]Liu W, Liu Y, Zhu J, Wright E, Ding I, Rodgers GP: Reduced hGC-1 protein expression is associated with malignant progression of colon carcinoma. Clin Cancer Res 2008, 14:1041-1049.
• [14]Conrotto P, Roesli C, Rybak J, Kischel P, Waltregny D, Neri D, Castronovo V: Identification of new accessible tumor antigens in human colon cancer by ex vivo protein biotinylation and comparative mass spectrometry analysis. Int J Cancer 2008, 123:2856-2864.
• [15]Chen L, Li H, Liu W, Zhu J, Zhao X, Wright E, Cao L, Ding I, Rodgers GP: Olfactomedin 4 suppresses prostate cancer cell growth and metastasis via negative interaction with cathepsin D and SDF-1. Carcinogenesis 2011, 32:986-994.
• [16]Liu W, Lee HW, Liu Y, Wang R, Rodgers GP: Olfactomedin 4 is a novel target gene of retinoic acids and 5-aza-2'-deoxycytidine involved in human myeloid leukemia cell growth, differentiation, and apoptosis. Blood 2010, 116:4938-4947.
• [17]Huang Y, Yang M, Yang H, Zeng Z: Upregulation of the GRIM-19 gene suppresses invasion and metastasis of human gastric cancer SGC-7901 cell line. Exp Cell Res 2010, 316:2061-2070.
• [18]Livak KJ, Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001, 25:402-408.
• [19]Yasui W, Oue N, Aung PP, Matsumura S, Shutoh M, Nakayama H: Molecular-pathological prognostic factors of gastric cancer: a review. Gastric Cancer 2005, 8:86-94.
• [20]Aung PP, Oue N, Mitani Y, Nakayama H, Yoshida K, Noguchi T, Bosserhoff AK, Yasui W: Systematic search for gastric cancer-specific genes based on SAGE data: melanoma inhibitory activity and matrix metaloproteinase-10 are novel prognostic factors in patients with gastric cancer. Oncogene 2006, 25:2546-2557.
• [21]Matsukura S, Jones PA, Takai D: Establishment of conditional vectors for hairpin siRNA knockdowns. Nucleic Acids Res 2003, 31:e77.
• [22]Altieri DC: Validating survivin as a cancer therapeutic target. Nat Rev Cancer 2003, 3:46-54.
• [23]Chene P: Inhibiting the p53-MDM2 interaction: an important target for cancer therapy. Nat Rev Cancer 2003, 3:102-109.
• [24]Ueda M, Kokura S, Imamoto E, Naito Y, Handa O, Takagi T, Yoshida N, Yoshikawa T: Blocking of NF-kappaB activation enhances the tumor necrosis factor alpha-induced apoptosis of a human gastric cancer cell line. Cancer Lett 2003, 193:177-182.
• [25]Kim KK, Park KS, Song SB, Kim KE: Up regulation of GW112 gene by NFkB promotes an antiapoptotic property in gastric cancer cells. Mol Carcinog 2010, 49:259-270.