Biomarker Commons

Rho GDP-dissociation inhibitor alpha is associated with cancer metastasis in colon and prostate cancer.

Pharmazie. 2012 Mar;67(3):253-5

Authors: Yamashita T, Okamura T, Nagano K, Imai S, Abe Y, Nabeshi H, Yoshikawa T, Yoshioka Y, Kamada H, Tsutsumi Y, Tsunoda S

Abstract
Since metastasis is one of the most important prognostic factors in colorectal cancer, development of new methods to diagnose and prevent metastasis is highly desirable. However, the molecular mechanisms leading to the metastatic phenotype have not been well elucidated. In this study, a proteomics-based search was carried out for metastasis-related proteins in colorectal cancer by analyzing the differential expression of proteins in primary versus metastasis focus-derived colorectal tumor cells. Protein expression profiles were determined using a tissue microarray (TMA), and the results identified Rho GDP-dissociation inhibitor alpha (Rho GDI) as a metastasis-related protein in colon and prostate cancer patients. Consequently, Rho GDI may be useful as a diagnostic biomarker and/or a therapeutic to prevent colon and prostate cancer metastasis.

PMID: 22530308 [PubMed - indexed for MEDLINE]

Serum vascular endothelial growth factor-C and lymphoangiogenesis are associated with the lymph node metastasis and prognosis of patients with colorectal cancer.

ANZ J Surg. 2011 Oct;81(10):694-9

Authors: Wang TB, Chen ZG, Wei XQ, Wei B, Dong WG

Abstract
BACKGROUND: The study aims to investigate the relationship among serum vascular endothelial growth factor (SVEGF-C), VEGF-C expression and lymph vessel density (LVD) in tumour tissue, and their influence to colorectal carcinoma (CRC).
METHODS: The SVEGF-C concentration of 110 patients with CRC and 40 healthy donors was examined by ELISA. The 110 tumour tissues and 40 normal colorectal specimens were examined by immunohistochemical staining (SP method) with VEGF-C and podoplanin (lymphatic vessel specific antibody). Kaplan–Meier survival analysis determined the influence on CRC prognosis.
RESULTS: CRC SVEGF-C level (889.0 ± 264.0 pg/mL) significantly exceeded (P = 0.000) the control level (373.2 ± 97.3 ng/L), and was significantly higher in T3, lymph node metastasis (LNM), distant metastasis, and pTNM groups III and IV. LNM prediction sensitivity, specificity, and accuracy of SVEGF-C were 85.7, 80.0 and 83.6%, respectively (875 pg/mL cut-off). VEGF-C expression was elevated in CRC versus control patients (P = 0.000), and was significantly related to LNM and pTNM stages III and IV. Mean LVD in CRC (6.3 ± 0.7/200 HP) significantly exceeded control mean (3.0 ± 0.7/200 HP) (P = 0.000). LVD was significantly higher in LNM and pTNM stages III and IV. SVEGF-C level was significantly higher in VEGF-C positive versus negative patients (P = 0.000), and was related to LVD (P = 0.009). Kaplan–Meier ranking of prognostic factors was SVEGF-C level (P = 0.000), VEGF-C expression (P = 0.001) and LVD (P = 0.012).
CONCLUSION: SVEGF-C level, VEGF-C and LVD are related to LNM and poor prognosis in patients with CRC. SVEGF-C may be a biomarker for LNM in CRC.

PMID: 22295309 [PubMed - indexed for MEDLINE]

miRNAs are stable in colorectal cancer archival tissue blocks.

Front Biosci (Elite Ed). 2012;4:1937-40

Authors: Bovell L, Shanmugam C, Katkoori VR, Zhang B, Vogtmann E, Grizzle WE, Manne U

Abstract
MicroRNAs (miRNAs) have prognostic and therapeutic value for colorectal cancers RCs). Although formalin-fixed paraffin-embedded (FFPE) tissues are available for biomarker studies, the stability of miRNAs in these tissues stored for long periods (more than 20 years) is unknown. The present effort involved analysis of 345 FFPE CRC tissues, stored for 6 to 28 years (1982-2004), for the expression of six miRNAs (miR-20a, miR-21, miR-106a, miR-181b, miR-203, and miR-324-5p) using TaqMan(r) microRNA assays and quantitative real-time PCR (qRT-PCR). Evaluation, by linear regression analysis, of miRNA expression among archived CRC tissues found similar levels of all six miRNAs in tissues stored over this period (correlation coefficients, R2, ranged from less than 0.0001-0.009; and t-test p-values were greater than or equal to 0.05). Thus, miRNAs are stable in FFPE tissues stored for long periods of time, and such samples can be used for discovery of biomarkers.

PMID: 22202009 [PubMed - indexed for MEDLINE]

Simultaneous multianalyte immunoassay measurement of five serum tumor markers in the detection of colorectal cancer.

Anticancer Res. 2012 Mar;32(3):985-8

Authors: Lumachi F, Marino F, Orlando R, Chiara GB, Basso SM

Abstract
Several serum tumor markers (STMs) have been proposed for the diagnosis of colorectal cancer (CRC), but their detection should be combined to increase accuracy. The measurement of a serum biomarker panel may improve the diagnostic value of single STM and a multianalyte immunoassay approach can shorten assay time and lower sample consumption. The aim of this study was to determine whether the simultaneous multianalyte immunoassay is useful for early detection of CRC. We measured a panel of five STMs namely, carcinoembryonic antigen (CEA), cancer antigen (CA) 19-9 and 72-4, cytokeratin fragment (CYFRA) 21-1, and osteopontin, in a selected homogeneous population of 102 consecutive patients (median age 66 years, range 42-75 years) with Dukes B, G1-2, colorectal adenocarcinoma (cases) and in a group of 99 age- and sex-matched patients suffering from confirmed benign colorectal diseases (controls). Overall, 141 (70.1%) men and 60 (29.9%) women were studied. The highest sensitivity was 45.1% (osteopontin), while the highest specificity was 90.9% (CEA). The accuracy was lower, ranging from 24.9% (CA 19-9) to 67.2% (CEA). CYFRA 21-1 and CA 72-4 had similar sensitivity (35.3% and 31.4%, respectively), but a significantly different specificity (37.4% vs. 89.9%). A combination of the five markers achieved 74.1% sensitivity and 94.3% specificity. In conclusion, in patients with CRC all single STMs show low sensitivity and specificity, while the simultaneous measurement of a panel of STMs may increase the diagnostic accuracy. When the sample volume is limited, the multianalyte immunoassay can be a reliable tool for studying patients undergoing laboratory screening for CRC.

PMID: 22399621 [PubMed - indexed for MEDLINE]

SEL1L, an UPR response protein, a potential marker of colonic cell transformation.

Dig Dis Sci. 2012 Apr;57(4):905-12

Authors: Ashktorab H, Green W, Finzi G, Sessa F, Nouraie M, Lee EL, Morgano A, Moschetta A, Cattaneo M, Mariani-Costantini R, Brim H, Biunno I

Abstract
BACKGROUND: SEL1L gene product is implicated in the endoplasmic reticulum (ER)-associated protein degradation and Unfolded Protein Response pathways. This gene and associated miRNAs have been indicated as predictive and prognostic markers of pancreatic cancer.
AIM: Explore the role of SEL1L in colorectal cancer (CRC) progression.
METHODS: SEL1L expression was analysed immunohistochemically in 153 adenomas and 71 CRCs from African American and North Italian patients. The distribution of stained cells was determined by computing median and inter quartile range. The receiver operating characteristics plot was used as discriminate power of SEL1L expression, CRC diagnosis and the effects on patient survival.
RESULTS: SEL1L was low in normal mucosa and confined to few scattered cells at the base crypt of the villi and in the foveolar glandular compartment. The highest levels were in Paneth cells within the lysosomes. The enterocytic progenitor cells and mature enterocytes showed less cytoplasmic staining. In CRCs, SEL1L expression significantly correlated with the progression from adenoma to carcinoma (P = 0.0001) being stronger in well-to-moderately differentiated cancers. No correlation was found with other clinicopathological characteristics or ethnicity.
CONCLUSIONS: SEL1L expression is a potential CRC tissue biomarker since its expression is significantly higher in adenoma cells with respect to normal mucosa. The levels of expression decrease sensibly in undifferentiated CRC cancers. Interestingly, Paneth cells contain high levels of SEL1L protein that could indicate pre-neoplastic mucosa undergoing neoplastic transformation. Since SEL1L's major function lies within ER stress and active ERAD response, it may identify CRCs with differentiated secretory phenotype and acute cellular stress.

PMID: 22350780 [PubMed - indexed for MEDLINE]

DNA methylation biomarker candidates for early detection of colon cancer.

Tumour Biol. 2012 Apr;33(2):363-72

Authors: Yi JM, Dhir M, Guzzetta AA, Iacobuzio-Donahue CA, Heo K, Yang KM, Suzuki H, Toyota M, Kim HM, Ahuja N

Abstract
Promoter CpG island hypermethylation of tumor suppressor genes is a common hallmark of all human cancers. Many researchers have been looking for potential epigenetic therapeutic targets in cancer using gene expression profiling with DNA microarray approaches. Our recent genome-wide platform of CpG island hypermethylation and gene expression in colorectal cancer (CRC) cell lines revealed that FBN2 and TCERG1L gene silencing is associated with DNA hypermethylation of a CpG island in the promoter region. In this study, promoter DNA hypermethylation of FBN2 and TCERG1L in CRC occurs as an early and cancer-specific event in colorectal cancer. Both genes showed high frequency of methylation in colon cancer cell lines (>80% for both of genes), adenomas (77% for FBN2, 90% for TCERG1L, n = 39), and carcinomas (86% for FBN2, 99% for TCERG1L, n = 124). Bisulfite sequencing confirmed cancer-specific methylation of FBN2 and TCERG1L of promoters in colon cancer cell line and cancers but not in normal colon. Methylation of FBN2 and TCERG1L is accompanied by downregulation in cell lines and in primary tumors as described in the Oncomine™ website. Together, our results suggest that gene silencing of FBN2 and TCERG1L is associated with promoter DNA hypermethylation in CRC tumors and may be excellent biomarkers for the early detection of CRC.

PMID: 22238052 [PubMed - indexed for MEDLINE]

Defective mismatch repair status as a prognostic biomarker of disease-free survival in stage III colon cancer patients treated with adjuvant FOLFOX chemotherapy.

Clin Cancer Res. 2011 Dec 1;17(23):7470-8

Authors: Zaanan A, Fléjou JF, Emile JF, Des GG, Cuilliere-Dartigues P, Malka D, Lecaille C, Validire P, Louvet C, Rougier P, de Gramont A, Bonnetain F, Praz F, Taïeb J

Abstract
PURPOSE: Adding oxaliplatin to adjuvant 5-fluorouracil (5-FU) chemotherapy improves 3-year disease-free survival (DFS) after resection of stage III colon cancer. Several studies suggest that patients with tumors exhibiting defective mismatch repair (MMR) do not benefit from adjuvant 5-FU chemotherapy, but there are few data on 5-FU-oxaliplatin (FOLFOX) adjuvant chemotherapy in this setting. The aim of this study was to evaluate the prognostic value of MMR status for DFS in patients with stage III colon cancer receiving adjuvant FOLFOX chemotherapy.
EXPERIMENTAL DESIGN: MMR status was determined by microsatellite instability testing or immunohistochemistry in 303 unselected patients with stage III colon cancer receiving adjuvant FOLFOX chemotherapy in 9 centers. Cox proportional hazards models were used to examine the association between MMR status and 3-year DFS.
RESULTS: The 3-year DFS rate was significantly higher in the 34 patients (11.2% of the study population) with defective MMR tumors (90.5%) than in patients with proficient MMR tumors (73.8%; log-rank test; HR = 2.16; 95% CI, 1.09-4.27; P = 0.027). In multivariate analysis, MMR status remained an independent significant prognostic factor for DFS (HR = 4.48; 95% CI, 1.34-14.99; P = 0.015).
CONCLUSION: MMR status is an independent prognostic biomarker for DFS in patients with stage III colon cancer receiving adjuvant FOLFOX chemotherapy.

PMID: 21998335 [PubMed - indexed for MEDLINE]

HIWI expression profile in cancer cells and its prognostic value for patients with colorectal cancer.

Chin Med J (Engl). 2011 Jul;124(14):2144-9

Authors: Zeng Y, Qu LK, Meng L, Liu CY, Dong B, Xing XF, Wu J, Shou CC

Abstract
BACKGROUND: HIWI is a member of PIWI gene family and its expression is found in various tumors, indicating that it may play a pivotal role in tumor development. This study was designated to examine HIWI protein expression profile in several cancer cell lines and its prognostic value for patients with colorectal cancer.
METHODS: Totally 270 patients who underwent surgical resection of primary colorectal cancer between January 1999 and December 2002 with a median follow-up time of 33 months were registered in the study. Formalin-fixed and paraffin-embedded specimens from these patients and 236 matched adjacent non-cancerous normal colorectal tissues were collected. Anti-HIWI monoclonal antibodies were generated and used for evaluating HIWI protein expression. χ(2) tests were conducted to determine the association between HIWI expression and the other variables. Survival curves were estimated using the Kaplan-Meier method and compared by the log-rank test. Multivariate analysis was performed by using the Cox regression model.
RESULTS: By generating antibodies specific for HIWI, we examined HIWI protein expression in several cancer cell lines and demonstrated positive expression of HIWI in 69 out of 270 (25.6%) colorectal cancer tissues; 15 of 236 (6.4%) matched adjacent non-cancerous tissues were also positive for HIWI. Patients with positive HIWI expression in adjacent non-cancerous tissue had statistically lower overall survival (OS) and disease free survival (DFS) compared with negative patients (OS: 10.4% vs. 55.5%, P = 0.009; DFS: 10.4% vs. 55.1%, P = 0.015). For early stage group (stages I and II), patients with positive HIWI expression had significantly lower OS and DFS (OS: 57.4% vs. 79.5%, P = 0.014; DFS: 56.7% vs. 80.5%, P = 0.010). In lymph node negative group, patients with positive HIWI expression had statistically lower OS and DFS (OS: 53.0% vs. 73.5%, P = 0.037; DFS: 52.2% vs. 74.6%, P = 0.025). Multivariate analysis revealed that HIWI over-expression was a significant prognostic factor for OS (95%CI: 1.132 - 2.479, P = 0.010).
CONCLUSION: HIWI could be a potential prognostic biomarker for the patients with colorectal cancer, especially for those at early stages or without lymph node metastasis.

PMID: 21933617 [PubMed - indexed for MEDLINE]

Histone deacetylase inhibition increases levels of choline kinase α and phosphocholine facilitating noninvasive imaging in human cancers.

Cancer Res. 2012 Feb 15;72(4):990-1000

Authors: Beloueche-Babari M, Arunan V, Troy H, te Poele RH, te Fong AC, Jackson LE, Payne GS, Griffiths JR, Judson IR, Workman P, Leach MO, Chung YL

Abstract
Histone deacetylase (HDAC) inhibitors are currently approved for cutaneous T-cell lymphoma and are in mid-late stage trials for other cancers. The HDAC inhibitors LAQ824 and SAHA increase phosphocholine (PC) levels in human colon cancer cells and tumor xenografts as observed by magnetic resonance spectroscopy (MRS). In this study, we show that belinostat, an HDAC inhibitor with an alternative chemical scaffold, also caused a rise in cellular PC content that was detectable by (1)H and (31)P MRS in prostate and colon carcinoma cells. In addition, (1)H MRS showed an increase in branched chain amino acid and alanine concentrations. (13)C-choline labeling indicated that the rise in PC resulted from increased de novo synthesis and correlated with an induction of choline kinase α expression. Furthermore, metabolic labeling experiments with (13)C-glucose showed that differential glucose routing favored alanine formation at the expense of lactate production. Additional analysis revealed increases in the choline/water and phosphomonoester (including PC)/total phosphate ratios in vivo. Together, our findings provide mechanistic insights into the impact of HDAC inhibition on cancer cell metabolism and highlight PC as a candidate noninvasive imaging biomarker for monitoring the action of HDAC inhibitors.

PMID: 22194463 [PubMed - indexed for MEDLINE]

Development and independent validation of a prognostic assay for stage II colon cancer using formalin-fixed paraffin-embedded tissue.

J Clin Oncol. 2011 Dec 10;29(35):4620-6

Authors: Kennedy RD, Bylesjo M, Kerr P, Davison T, Black JM, Kay EW, Holt RJ, Proutski V, Ahdesmaki M, Farztdinov V, Goffard N, Hey P, McDyer F, Mulligan K, Mussen J, O'Brien E, Oliver G, Walker SM, Mulligan JM, Wilson C, Winter A, O'Donoghue D, Mulcahy H, O'Sullivan J, Sheahan K, Hyland J, Dhir R, Bathe OF, Winqvist O, Manne U, Shanmugam C, Ramaswamy S, Leon EJ, Smith WI, McDermott U, Wilson RH, Longley D, Marshall J, Cummins R, Sargent DJ, Johnston PG, Harkin DP

Abstract
PURPOSE: Current prognostic factors are poor at identifying patients at risk of disease recurrence after surgery for stage II colon cancer. Here we describe a DNA microarray-based prognostic assay using clinically relevant formalin-fixed paraffin-embedded (FFPE) samples.
PATIENTS AND METHODS: A gene signature was developed from a balanced set of 73 patients with recurrent disease (high risk) and 142 patients with no recurrence (low risk) within 5 years of surgery.
RESULTS: The 634-probe set signature identified high-risk patients with a hazard ratio (HR) of 2.62 (P < .001) during cross validation of the training set. In an independent validation set of 144 samples, the signature identified high-risk patients with an HR of 2.53 (P < .001) for recurrence and an HR of 2.21 (P = .0084) for cancer-related death. Additionally, the signature was shown to perform independently from known prognostic factors (P < .001).
CONCLUSION: This gene signature represents a novel prognostic biomarker for patients with stage II colon cancer that can be applied to FFPE tumor samples.

PMID: 22067406 [PubMed - indexed for MEDLINE]

From discovery to the clinic: the novel DNA methylation biomarker (m)SEPT9 for the detection of colorectal cancer in blood.

Epigenomics. 2010 Aug;2(4):575-85

Authors: Payne SR

Abstract
Detection of colorectal cancer at an early stage has been shown to significantly decrease mortality from the disease, while the advent of effective therapies for late-stage colorectal cancer make the detection of colorectal cancer at any stage a critical step in further reducing colorectal cancer mortality. Availability of a blood-based test for colorectal cancer is expected to improve screening compliance in the general population. Through DNA methylation-sensitive, restriction enzyme-based biomarker discovery, we identified a region of the Septin 9 gene that is methylated in over 90% of colorectal cancer tissues with little or no methylation seen in normal colon tissue and other controls. Specific detection of colorectal cancer DNA using the Septin 9 methylation biomarker ((m)SEPT9) was demonstrated in multiple studies of plasma from colorectal cancer patients and colonoscopy-verified negative controls. A prospective, population-based trial to determine the clinical performance of (m)SEPT9 in colorectal cancer screening guideline-eligible individuals has recently been completed, with the results to be published in the near future. The potential pitfalls and lessons learned in the multiyear process of developing the (m)SEPT9 biomarker from initial discovery to commercialization are described in this article.

PMID: 22121975 [PubMed - indexed for MEDLINE]

Specific hypomethylated CpGs at the IGF2 locus act as an epigenetic biomarker for familial adenomatous polyposis colorectal cancer.

Epigenomics. 2010 Jun;2(3):365-75

Authors: Miroglio A, Jammes H, Tost J, Ponger L, Gut IG, El Abdalaoui H, Coste J, Chaussade S, Arimondo PB, Lamarque D, Dandolo L

Abstract
AIMS: The identification of specific biomarkers for colorectal cancer is of primary importance for early diagnosis. The aim of this study was to evaluate if methylation changes at the IGF2/H19 locus could be predictive for individuals at high risk for developing sporadic or hereditary colorectal cancer.
MATERIALS &#38; METHODS: Quantitative methylation analysis using pyrosequencing was performed on three differentially methylated regions (DMRs): IGF2 DMR0 and DMR2 and the H19 DMR in DNA samples from sporadic colorectal cancer (n = 26), familial adenomatous polyposis (n = 35) and hereditary nonpolyposis colorectal cancer (n = 19) patients.
RESULTS: We report in this article for the first time, that in sporadic colorectal cancer tumor DNA both the IGF2 DMR0 and DMR2 are hypomethylated, while the H19 DMR retains its monoallelic methylation pattern. In lymphocyte DNA, a striking hypomethylation of nine contiguous correlated CpGs was found in the IGF2 DMR2 but only in familial adenomatous polyposis patients.
CONCLUSION: Methylation alterations at the IGF2 locus are more extensive than previously reported and DMR2 hypomethylation in lymphocyte DNA might be a specific epigenetic biomarker for familial adenomatous polyposis patients.

PMID: 22121898 [PubMed - indexed for MEDLINE]

Intensive radiographic and biomarker surveillance in stage II and III colorectal cancer.

Oncology. 2012;82(1):41-7

Authors: Fora A, Patta A, Attwood K, Wilding G, Fakih M

Abstract
OBJECTIVE: The objective of this study was to determine the rate of salvage resections in patients with stage II and III colorectal cancer following intensive surveillance in a comprehensive cancer center.
METHODS: Patients with stage II and III colorectal cancer with a minimum follow-up of 3 years were included. Carcinoembryonic antigen was obtained every 3 months for 2 years and then every 6 months for years 3-5. CT scans of the chest, abdomen and pelvis were performed every 6 months for 2 years and then yearly for years 3-5. Colonoscopy was performed at year 1 and then every 3 years.
RESULTS: One hundred and seventy-seven patients were followed for a median of 60 months; 44 patients were diagnosed with recurrent disease. CT was the first sign of recurrence in 68% of patients. Carcinoembryonic antigen test was normal in 20 patients (45%) at the time of disease recurrence. Twenty-five patients (57%) with recurrent disease underwent curative-intent resection, 12 of whom are still cancer free at a median follow-up of 81 months.
CONCLUSIONS: In this retrospective study, intensive radiographic screening was associated with a high salvage resection rate, which led to favorable clinical outcomes. Randomized clinical trials are urgently needed to define the optimal duration and schedule of radiographic screening in stage II and III colorectal cancer.

PMID: 22286543 [PubMed - indexed for MEDLINE]

Dietary intake of PUFAs and colorectal polyp risk.

Am J Clin Nutr. 2012 Mar;95(3):703-12

Authors: Murff HJ, Shrubsole MJ, Cai Q, Smalley WE, Dai Q, Milne GL, Ness RM, Zheng W

Abstract
BACKGROUND: Marine-derived n-3 (omega-3) PUFAs may reduce risk of developing colorectal cancer; however, few studies have investigated the association of n-3 PUFA intakes on colorectal polyp risk.
OBJECTIVE: The objective of this study was to examine the associations of dietary PUFA intake on risk of colorectal adenomatous and hyperplastic polyps.
DESIGN: This was a colonoscopy-based case-control study that included 3166 polyp-free control subjects, 1597 adenomatous polyp cases, and 544 hyperplastic polyp cases. Dietary PUFA intake was calculated from food-frequency questionnaires and tested for association by using unconditional logistic regression. The urinary prostaglandin E(2) metabolite, which is a biomarker of prostaglandin E(2) production, was measured in 896 participants by using liquid chromatography and tandem mass spectrometry.
RESULTS: n-6 PUFAs were not associated with adenomatous or hyperplastic polyps in either men or women. Marine-derived n-3 PUFAs were associated with reduced risk of colorectal adenomas in women only, with an adjusted OR of 0.67 (95% CI: 0.47, 0.97) for the highest quintile of intake compared with the lowest quintile of intake (P-trend = 0.01). Dietary intake of α-linolenic acid was associated with an increased risk of hyperplastic polyps in men (P-trend = 0.03), which was not seen in women. In women, but not in men, dietary intake of marine-derived n-3 PUFAs was negatively correlated with urinary prostaglandin E(2) production (r = -0.18; P = 0.002).
CONCLUSION: Higher intakes of marine-derived n-3 PUFAs are associated with lower risk of adenomatous polyps in women, and the association may be mediated in part through a reduction in the production of prostaglandin E(2). This trial was registered at clinicaltrials.gov as NCT00625066.

PMID: 22277551 [PubMed - indexed for MEDLINE]

Analysis of the RUNX3 gene methylation in serum DNA from esophagus squamous cell carcinoma, gastric and colorectal adenocarcinoma patients.

Hepatogastroenterology. 2011 Nov-Dec;58(112):2007-11

Authors: Zheng Y, Zhang Y, Huang X, Chen L

Abstract
BACKGROUND/AIMS: Human runt-related transcription factor 3 (RUNX3) gene was a new candidate tumor suppressor gene involved in the TGF-β signaling pathway. We evaluated the diagnostic role of RUNX3 gene methylation in serum DNA of esophageal squamous cell carcinoma (ESCC), gastric cancer (GC) and colorectal cancer (CRC) patients.
METHODOLOGY: Methylation-specific polymerase chain reaction (MSP) was used to determine the promoter methylation status of RUNX3 gene in serum DNA. The combination of RUNX3 hypermethylation and conventional tumor markers was further analyzed.
RESULTS: Hypermethylation of RUNX3 was detected in 51.4% (36/70) of ESCC, 45.2% (28/62) of GC and 41.5% (27/65) of CRC patients, which was significantly higher than that of benign gastrointestinal diseases and healthy donors (p<0.001). Interestingly, aberrant RUNX3 methylation in serum DNA was associated with advanced stage (p=0.027) and lymph metastasis (p=0.032) in ESCC, but not in GC and CRC. Furthermore, the combinational analysis of CEA, CA19-9 and RUNX3 methylation showed a higher sensitivity and no reduced diagnostic specificity than CEA and CA19-9 combination in the three cancers.
CONCLUSIONS: The serum RUNX3 promoter hypermethylation may be a promising biomarker for the early diagnosis of ESCC, GC and CRC, which was further confirmed by combining with CEA and CA19-9.

PMID: 22234069 [PubMed - indexed for MEDLINE]

The proteomics of colorectal cancer: identification of a protein signature associated with prognosis.

PLoS One. 2011;6(11):e27718

Authors: O'Dwyer D, Ralton LD, O'Shea A, Murray GI

Abstract
Colorectal cancer is one of the commonest types of cancer and there is requirement for the identification of prognostic biomarkers. In this study protein expression profiles have been established for colorectal cancer and normal colonic mucosa by proteomics using a combination of two dimensional gel electrophoresis with fresh frozen sections of paired Dukes B colorectal cancer and normal colorectal mucosa (n = 28), gel image analysis and high performance liquid chromatography-tandem mass spectrometry. Hierarchical cluster analysis and principal components analysis showed that the protein expression profiles of colorectal cancer and normal colonic mucosa clustered into distinct patterns of protein expression. Forty-five proteins were identified as showing at least 1.5 times increased expression in colorectal cancer and the identity of these proteins was confirmed by liquid chromatography-tandem mass spectrometry. Fifteen proteins that showed increased expression were validated by immunohistochemistry using a well characterised colorectal cancer tissue microarray containing 515 primary colorectal cancer, 224 lymph node metastasis and 50 normal colonic mucosal samples. The proteins that showed the greatest degree of overexpression in primary colorectal cancer compared with normal colonic mucosa were heat shock protein 60 (p<0.001), S100A9 (p<0.001) and translationally controlled tumour protein (p<0.001). Analysis of proteins individually identified 14-3-3β as a prognostic biomarker (χ² = 6.218, p = 0.013, HR = 0.639, 95%CI 0.448-0.913). Hierarchical cluster analysis identified distinct phenotypes associated with survival and a two-protein signature consisting of 14-3-3β and aldehyde dehydrogenase 1 was identified as showing prognostic significance (χ² = 7.306, p = 0.007, HR = 0.504, 95%CI 0.303-0.838) and that remained independently prognostic (p = 0.01, HR = 0.416, 95%CI 0.208-0.829) in a multivariate model.

PMID: 22125622 [PubMed - indexed for MEDLINE]

A quantitative proteomic approach of the different stages of colorectal cancer establishes OLFM4 as a new nonmetastatic tumor marker.

Mol Cell Proteomics. 2011 Dec;10(12):M111.009712

Authors: Besson D, Pavageau AH, Valo I, Bourreau A, Bélanger A, Eymerit-Morin C, Moulière A, Chassevent A, Boisdron-Celle M, Morel A, Solassol J, Campone M, Gamelin E, Barré B, Coqueret O, Guette C

Abstract
Expression profiles represent new molecular tools that are useful to characterize the successive steps of tumor progression and the prediction of recurrence or chemotherapy response. In this study, we have used quantitative proteomic analysis to compare different stages of colorectal cancer. A combination of laser microdissection, OFFGEL separation, iTRAQ labeling, and MALDI-TOF/TOF MS was used to explore the proteome of 28 colorectal cancer tissues. Two software packages were used for identification and quantification of differentially expressed proteins: Protein Pilot and iQuantitator. Based on ∼1,190,702 MS/MS spectra, a total of 3138 proteins were identified, which represents the largest database of colorectal cancer realized to date and demonstrates the value of our quantitative proteomic approach. In this way, individual protein expression and variation have been identified for each patient and for each colorectal dysplasia and cancer stage (stages I-IV). A total of 555 proteins presenting a significant fold change were quantified in the different stages, and this differential expression correlated with immunohistochemistry results reported in the Human Protein Atlas database. To identify a candidate biomarker of the early stages of colorectal cancer, we focused our study on secreted proteins. In this way, we identified olfactomedin-4, which was overexpressed in adenomas and in early stages of colorectal tumors. This early stage overexpression was confirmed by immunohistochemistry in 126 paraffin-embedded tissues. Our results also indicate that OLFM4 is regulated by the Ras-NF-κB2 pathway, one of the main oncogenic pathways deregulated in colorectal tumors.

PMID: 21986994 [PubMed - indexed for MEDLINE]

Investigation of the prevalence and number of aberrant crypt foci associated with human colorectal neoplasm.

Cancer Epidemiol Biomarkers Prev. 2011 Sep;20(9):1918-24

Authors: Sakai E, Takahashi H, Kato S, Uchiyama T, Hosono K, Endo H, Maeda S, Yoneda M, Taguri M, Nakajima A

Abstract
BACKGROUND: Aberrant crypt foci (ACF) are considered to be useful as surrogate biomarker for colorectal cancer (CRC), but the biological significance of ACF remains controversial. We attempted to investigate the relationship between the presence of ACF and human colorectal carcinogenesis using a relatively large sample size.
METHODS: We carried out high-magnification chromoscopic colonoscopy to identify ACFs in 861 subjects undergoing a diagnostic endoscopy at the Yokohama City University Hospital. The present study compared the prevalence and number of ACFs in three subject groups (normal subjects, adenoma cases, and CRC cases). The correlations between the demographic and behavioral characteristics of the subjects and the prevalence of ACFs were also assessed.
RESULTS: The prevalence of ACF was 64%, 88%, and 95%, and the mean number of ACF was 3.6, 6.2, and 10.1, in normal subjects, adenoma cases, and CRC cases, respectively. When differences in the prevalence and number of ACFs among age- and sex-stratified subject groups were examined, significant stepwise increments from normal subjects to adenoma cases to CRC cases were apparent (P < 0.001). Moreover, an age- and sex-adjusted multiple logistic regression analysis revealed that smoking and alcohol habits had a synergistic effect, increasing the prevalence of ACFs as well as the risk of CRC (P < 0.001).
CONCLUSIONS: These results suggested that ACF may serve as a reliable surrogate biomarker for human colorectal carcinogenesis.
IMPACT: The use of ACF as an endpoint may enable the size, duration, and cost of CRC chemoprevention studies to be reduced.

PMID: 21750169 [PubMed - indexed for MEDLINE]

No association of the eNOS gene polymorphisms with survival in patients with colorectal cancer.

Med Oncol. 2011 Dec;28(4):1075-9

Authors: Kim YJ, Lee SJ, Kim JG, Sohn SK, Chae YS, Moon JH, Kang BW, Park JY, Park JS, Choi GS

Abstract
Endothelial nitric oxide synthase (eNOS)-derived nitric oxide (NO) is involved in numerous physiologic and pathophysiologic process including tumor angiogenesis and apoptosis. Accordingly, the present study analyzed polymorphisms of eNOS gene and their impact on the prognosis for patients with colorectal cancer. Four hundred and forty-four consecutive patients with surgically resected colorectal adenocarcinoma were enrolled in the present study. The genomic DNA was extracted from fresh colorectal tissue and 2 polymorphisms of eNOS gene (eNOS T786C and eNOS G894T) determined using a real-time PCR genotyping assay. The 2 eNOS gene polymorphisms were successfully amplified, and the frequencies of each genotype are as follows [T786C: TT (82.2%), TC (16.9%), CC (0.9%); G894T: GG (82.0%), GT (17.3%), TT (0.7%)]. Multivariate survival analysis including stage, age, site of disease, and CEA level showed that these polymorphisms were not associated with survival. For the clinicopathologic parameters, CEA level and TNM stage were significant prognostic factors in a Cox model for survival. The eNOS gene polymorphisms investigated in this study were not found to be an independent prognostic marker for Korean patients with surgically resected colorectal cancer. However, further studies are warranted to clarify the role of the eNOS gene polymorphisms as a prognostic biomarker for colorectal patients with cancer.

PMID: 20706876 [PubMed - indexed for MEDLINE]

No association of the hypoxia-inducible factor-1α gene polymorphisms with survival in patients with colorectal cancer.

Med Oncol. 2011 Dec;28(4):1032-7

Authors: Lee SJ, Kim JG, Sohn SK, Chae YS, Moon JH, Kang BW, Park JS, Park JY, Choi GS

Abstract
Hypoxia-inducible factor-1 (HIF-1) is the key regulator of cellular response to hypoxia and presumably plays a central role in the control of tumor growth. The present study analyzed polymorphisms of HIF-1α gene and their impact on the prognosis for patients with colorectal cancer. Four hundred and forty-five consecutive patients with surgically resected colorectal adenocarcinoma were enrolled in the present study. The genomic DNA was extracted from fresh colorectal tissue, and 2 polymorphisms of HIF-1α gene (HIF-1α C1772T and HIF-1α G1790A) determined using a real-time PCR genotyping assay. The 2 HIF-1α gene polymorphisms were successfully amplified, and the frequencies of each genotype are as follows: [C1772T: CC (92.1%), CT (7.9%); G1790A: GG (93.0%), GA (7.0%)]. Survival analysis including stage, age, site of disease, and CEA level showed that these polymorphisms were not associated with survival. For the clinicopathologic parameters, CEA level and TNM stage were significant prognostic factors in a Cox model for survival. HIF-1α gene polymorphisms investigated in this study were not found to be an independent prognostic marker for Korean patients with surgically resected colorectal cancer. However, further studies are warranted to clarify the role of HIF-1α gene polymorphisms as a prognostic biomarker for colorectal cancer patients.

PMID: 20635169 [PubMed - indexed for MEDLINE]