Overexpression of SPARC gene in human gastric carcinoma and its clinic–pathologic significance

Gastric cancer is the second most common cancer in the world and the fifth leading cause of cancer-related death in Taiwan. To improve the survival of gastric cancer patients, biomarkers for early detection and effective anticancer therapy are required. An essential first step is to profile gene expression in gastric cancer and identify genes that are aberrantly expressed, and to do this cDNA microarrays were performed. The clinic–pathologic correlation and prognostic significance of the aberrantly expressed genes were evaluated to identify novel biomarkers of gastric cancer. Fresh surgical samples of tumour tissue and matching noncancerous mucosa were obtained immediately after gastric resection in 43 patients. Secreted Protein, Acidic and Rich in Cysteine (SPARC) (Osteonectins), one of the most highly expressed genes in both intestinal and diffuse gastric cancers in our microarray results, was selected for further study. The overexpression of SPARC was verified using real-time quantitative-reverse transcription–polymerase chain reaction (Q-RT–PCR), Northern blot and immunohistochemical staining. The expression of SPARC in tumour tissues was, on average, 4.27-fold increased (95% CI 2.68–5.85) compared to adjacent noncancerous mucosa (P<0.001). The expression of SPARC was higher in advanced (T2, T3 and T4) cancer compared to the early (T1) cancer (P=0.048) with regard to depth of wall invasion. Higher expression of SPARC was significantly associated with lymph node metastasis (P<0.001), lymphatic invasion (P=0.004) and perineural invasion (P=0.047). Expression of SPARC in patients in stage II and above was significantly higher than those in stage I (P=0.017). The 3-year survival of patients with lower expression of SPARC was significantly better than those with a higher expression (log rank P=0.047). These data indicate the potential of SPARC as a prognostic marker for gastric cancer.

[1]  Lajos Pusztai,et al.  Clinical application of cDNA microarrays in oncology. , 2003, The oncologist.

[2]  G. Zweiger,et al.  Knowledge discovery in gene-expression-microarray data: mining the information output of the genome. , 1999, Trends in biotechnology.

[3]  Japanese Gastric Cancer Association.  Japanese Classification of Gastric Carcinoma – 2nd English Edition – , 1998, Gastric Cancer.

[4]  E. Sage,et al.  SPARC (BM-40, Osteonectin) Inhibits the Mitogenic Effect of Vascular Endothelial Growth Factor on Microvascular Endothelial Cells* , 1998, The Journal of Biological Chemistry.

[5]  Yoshiyuki Sakaki,et al.  Osteonectin-expressing cells in human stomach cancer and their possible clinical significance. , 2002, Cancer letters.

[6]  K Motamed,et al.  SPARC (osteonectin/BM-40). , 1999, The international journal of biochemistry & cell biology.

[7]  M. Folkman,et al.  Inhibition of endothelial cell proliferation by SPARC is mediated through a Ca2+‐binding EF‐hand sequence , 1995, Journal of cellular biochemistry.

[8]  A. Miyauchi,et al.  Gene expression profiles in thyroid carcinomas , 2000, British Journal of Cancer.

[9]  A. Gown,et al.  Alterations in SPARC and VEGF immunoreactivity in epithelial ovarian cancer. , 2000, Gynecologic oncology.

[10]  E. Thompson,et al.  Doxycycline-Inducible Expression of SPARC/ Osteonectin/ BM40 in MDA-MB-231 Human Breast Cancer Cells Results in Growth Inhibition , 2002, Breast Cancer Research and Treatment.

[11]  P. Bornstein,et al.  Matricellular proteins: extracellular modulators of cell function. , 2002, Current opinion in cell biology.

[12]  P. Laurén.  Histogenesis of intestinal and diffuse types of gastric carcinoma. , 1991, Scandinavian journal of gastroenterology. Supplement.

[13]  C C Howe,et al.  Disruption of the Sparc locus in mice alters the differentiation of lenticular epithelial cells and leads to cataract formation. , 1999, Experimental eye research.

[14]  Yi-Hsin Wu,et al.  Activation of Antimetastatic Nm23-H1 Gene Expression by Estrogen and Its α-Receptor. , 2002, Endocrinology.

[15]  M. Rhyu,et al.  Genetic classification of intestinal-type and diffuse-type gastric cancers based on chromosomal loss and microsatellite instability , 2003, Virchows Archiv.

[16]  T. Hwang,et al.  Benefits of palliative surgery for far-advanced gastric cancer. , 2002, Chang Gung medical journal.

[17]  T. Krieg,et al.  BM-40 (osteonectin, SPARC) is expressed both in the epidermal and in the dermal compartment of adult human skin. , 1998, The Journal of investigative dermatology.

[18]  P. Chambon,et al.  Neoplastic progression of human colorectal cancer is associated with overexpression of the stromelysin‐3 and BM‐40/SPARC genes , 1995, International journal of cancer.

[19]  M. Young,et al.  Osteonectin/SPARC/BM-40 in human decidua and carcinoma, tissues characterized by de novo formation of basement membrane. , 1988, The American journal of pathology.

[20]  R L Vessella,et al.  Differential expression of osteonectin/SPARC during human prostate cancer progression. , 2000, Clinical cancer research : an official journal of the American Association for Cancer Research.

[21]  S. Hsu,et al.  Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. , 1981, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[22]  D. Ransohoff.  Developing Molecular Biomarkers for Cancer , 2003, Science.

[23]  Hiroki Kuniyasu,et al.  Molecular diagnosis of gastric cancer: present and future , 2001, Gastric Cancer.

[24]  B. le Bail,et al.  Osteonectin/SPARC is overexpressed in human hepatocellular carcinoma , 1999, The Journal of pathology.

[25]  M. Yao,et al.  SPARC expression in primary human renal cell carcinoma: upregulation of SPARC in sarcomatoid renal carcinoma. , 2001, Human pathology.

[26]  Yoshiyuki Sakaki,et al.  Appearance of Osteonectin‐expressing Fibroblastic Cells in Early Rat Stomach Carcinogenesis and Stomach Tumors Induced with N‐Methyl‐N′‐nitro‐N‐nitrosoguanidine , 2002, Japanese journal of cancer research : Gann.

[27]  H. Kanayama,et al.  Analysis of the gene expression of SPARC and its prognostic value for bladder cancer. , 2001, The Journal of urology.

[28]  F. Roviello,et al.  Prognostic Significance of CEA, CA 19-9 and CA 72-4 Preoperative Serum Levels in Gastric Carcinoma , 1999, Oncology.

[29]  R. Barreto-Zúñiga,et al.  Significance of Helicobacter pylori infection as a risk factor in gastric cancer: serological and histological studies. , 1997, Journal of gastroenterology.

[30]  L. Sobin,et al.  TNM classification of malignant tumors, fifth edition (1997) , 1997, Cancer.

[31]  M. Mori,et al.  Clinical significance of secreted protein acidic and rich in cystein in esophageal carcinoma and its relation to carcinoma progression , 2003, Cancer.

[32]  E. Tahara.  Molecular mechanism of stomach carcinogenesis , 2005, Journal of Cancer Research and Clinical Oncology.

[33]  O. Podhajcer,et al.  Suppression of SPARC expression by antisense RNA abrogates the tumorigenicity of human melanoma cells , 1997, Nature Medicine.

[34]  Yi-Hsin Wu,et al.  Activation of antimetastatic Nm23-H1 gene expression by estrogen and its alpha-receptor. , 2002, Endocrinology.

[35]  E. Sage,et al.  SPARC participates in the branching morphogenesis of developing fetal rat lung. , 1995, American journal of respiratory cell and molecular biology.

[36]  D. Massi,et al.  Osteonectin expression correlates with clinical outcome in thin cutaneous malignant melanomas. , 1999, Human pathology.

[37]  David F Ransohoff,et al.  Cancer. Developing molecular biomarkers for cancer. , 2003, Science.

[38]  Ronald W. Davis,et al.  Quantitative Monitoring of Gene Expression Patterns with a Complementary DNA Microarray , 1995, Science.

[39]  S. Rempel,et al.  A study of SPARC and vitronectin localization and expression in pediatric and adult gliomas: high SPARC secretion correlates with decreased migration on vitronectin. , 2000, International journal of oncology.

[40]  Z. Werb,et al.  SPARC, a secreted protein associated with morphogenesis and tissue remodeling, induces expression of metalloproteinases in fibroblasts through a novel extracellular matrix-dependent pathway , 1993, The Journal of cell biology.

[41]  O. Beahrs.  The American Joint Committee on Cancer. , 1984, Bulletin of the American College of Surgeons.

[42]  T. Nakajima,et al.  [Prognostic factors in gastric cancer]. , 1988, Gan to kagaku ryoho. Cancer & chemotherapy.

[43]  P. Brown,et al.  DNA arrays for analysis of gene expression. , 1999, Methods in enzymology.

[44]  M. Iruela-Arispe,et al.  SPARC is a source of copper-binding peptides that stimulate angiogenesis , 1994, The Journal of cell biology.

[45]  C. Iacobuzio-Donahue,et al.  The desmoplastic response to infiltrating breast carcinoma: gene expression at the site of primary invasion and implications for comparisons between tumor types. , 2002, Cancer research.

[46]  K. Dohi,et al.  Secreted protein acidic and rich in cysteine (SPARC) in patients with diabetic nephropathy and tubulointerstitial injury , 2000, Diabetologia.