Cancer Biomarkers - Volume 8, issue 2

Authors: Choschzick, Matthias | Heilenkötter, Uwe | Lebeau, Annette | Jaenicke, Fritz | Terracciano, Luigi | Bokemeyer, Carsten | Sauter, Guido | Simon, Ronald

Article Type: Research Article

Abstract: Background: MDM2 is overexpressed and amplified in a number of malignant tumors including breast carcinomas. Cell culture experiments showed a close connection between MDM2 expression and estrogen receptor status in breast cancer cell lines. Only little is known about the role of MDM2 amplifications in early-stage breast carcinomas with positive estrogen receptor status. Methods: 661 highly characterized node-negative breast carcinomas with positive estrogen receptor status (ER+ early-stage breast carcinomas) were analyzed on a tissue microarray. Molecular (HER2, CCND1, MDM2, MYC, 8q21), as well as estrogen receptor expression data used in this analysis, was available from previously published studies. …The primary endpoint of overall survival analysis was death after 10 years. Results: Gene amplifications were found in 194/661 (29%) ER+ early-stage breast carcinomas and 40 (7%) exhibited amplification of the MDM2 oncogene. MDM2 amplifications were significantly related to advanced tumor stage (p < 0.05) and high Ki67 expression levels (p < 0.05). There was no relationship between MDM2 copy number changes and tumor grade, estrogen receptor expression level and co-amplification of HER2, CCND1 and 8q. Tumor stage (pT1 vs pT2-pT4; HR 1.51; 95% CI 1.02–2.24; p=0.042), grading (G1-G2 vs G3; HR 2.27; 95% CI 1.51–3.43; p < 0.001), high Ki67 proliferation index (HR 2.03; 95% CI 1.31–3.15; p=0.0015), HER2 (HR 2.6; 95% CI 1.51 to 4.5; p < 0.001) and MDM2 amplification (HR 2.05, 95% CI 1.06–3.97, p =0.033) were statistically adverse prognostic risk factors in univariate Cox regression analysis. Patient age, estrogen receptor expression level, CCND1 and 8q amplification were not associated to overall survival. Multivariate Cox regression analysis of survival data included tumor stage, grading, Ki67 labeling index, HER2 and MDM2 amplification status. In this statistical model, only MDM2 amplification was an independent factor for overall patient survival in ER+ early- stage breast carcinomas (HR 2.64; 95% CI 1.32 to 5.28; p=0.006). Conclusion: The MDM2 oncogene is amplified in a substantial proportion of ER+ early-stage breast carcinomas and an independent parameter for poor patient outcome in this subgroup. The prognostic effect of MDM2 is closely connected to estrogen receptor expression of breast carcinomas. Show more

Keywords: MDM2, amplification, breast cancer, prognosis

DOI: 10.3233/DMA-2011-0806

Citation: Cancer Biomarkers, vol. 8, no. 2, pp. 53-60, 2011

Authors: Martinelli, Marcella | Ugolini, Giampaolo | Scapoli, Luca | Rivetti, Stefano | Lauriola, Mattia | Mattei, Gabriella | Rosati, Giancarlo | Montroni, Isacco | Manaresi, Alessio | Zattoni, Davide | Taffurelli, Mario | Solmi, Rossella

Article Type: Research Article

Abstract: Epidermal growth factor receptor (EGFR) family members (EGFR, HER2, HER3 and HER4) have been extensively investigated for its possible involvement in cancer development and progression. In colorectal cancer (CRC) EGFR family has been found frequently over-expressed, thus therapy targeting EGFR has been developed. Interestingly, it has been observed that genetic variants in these receptors may alter the therapeutic efficacy of EGFR inhibitors. Polymorphic variants in members of the EGFR family could influence different biologic activities, such as ligands affinity, dimerization efficiency, kinase activity, expression levels, with a consequent impact in signalling pathways and cell behaviour. This study aimed to verify …whether single nucleotide polymorphisms (SNPs) of EGFR family members could represent susceptibility factors able to influence the risk to develop CRC. Peripheral blood of 70 Italian colon cancer patients and 72 healthy controls was used as a source of genomic DNA to investigate EGFR, HER2 and HER3 common non-synonymous SNPs. Genetic association tests were performed to verify a possible relationship with CRC. Evidence of genotype association was found for the R521K EGFR polymorphism under a dominant mode of inheritance (Mid-P=0.031). Genotypes with the variant allele of EGFR R521K SNP confer a risk reduction to develop CRC. Show more

Keywords: Colorectal cancer, EGFR, HER2, HER3, polymorphisms

DOI: 10.3233/DMA-2011-0826

Citation: Cancer Biomarkers, vol. 8, no. 2, pp. 61-65, 2011

Authors: Liu, Jingeng | Hu, Yi | Hu, Wei | Xie, Xuan | ELA BELLA, Amos | Fu, Jianhua

Article Type: Research Article

Abstract: Inhibitor of differentiation 1 (Id1) plays a role in cellular differentiation, proliferation, angiogenesis and tumor invasion. In current study, we investigated the expression of Id1 protein in 189 specimens of stage III ESCC by immunohistochemistry. The correlation between Id1 expression and clinicopathological parameters in terms of gender, age, tumor location, differentiation, pathological T stage and pathological N stage was also explored. Immunohistochemical staining showed that Id1 was expressed in all cases (100%). According to ROC curve, we selected 6.33 as the cutoff score. There were 95 cases in high expression group (> 6.33) and 94 cases in low expression group …(⩽ 6.33) respectively. The Id1 expression was associated negatively with differentiation, and positively with pathological N stage (P< 0.05, bothly); No significant correlation was observed between Id1 expression and gender, age, tumor location or pathological T stage (P> 0.05). As shown by the Kaplan-Meier curve, the overall survival rate of high expression group was significantly lower than that of low expression group (P< 0.001). The expression of Id1 protein has a close relationship with differentiation degree, pathological N stage and survival in ESCC patients. Id1 could be considered as a prognostic predictor for stage III ESCC patients. Show more

Keywords: Esophageal neoplasm, Id1, immunohistochemistry, lymph node metastasis, prognosis

DOI: 10.3233/DMA-2011-0831

Citation: Cancer Biomarkers, vol. 8, no. 2, pp. 67-72, 2011

Authors: Zhao, Zeng-Ren | Zhang, Zhi-Yong | He, Xin-Qi | Hu, Yue-Ming | Tian, Yan-Feng | Zhang, Li-Jing | Sun, Xiao-Feng

Article Type: Research Article

Abstract: Objectives: We measured nucleoporin 88 (Nup88) mRNA expression in primary colorectal cancers to investigate its relationship with clinicopathological features and p53. Methods: The primary cancer tissues, adjacent noncancerous tissues and the proximal and distant margins of normal mucosa were collected from 73 colorectal cancer patients during surgery. Nup88 mRNA expression was measured on these fresh specimens and on colon cell lines HCT-116 p 53 + / + and HCT-116 p 53 − / − by RT-PCR while p53 mRNA and β -actin as controls. Nup88 and …p53 protein expression were then immunohistochemistrically examined in other 25 colorectal cancers specimens paraffin embedded and formalin fixed. Results: Nup88 expression was higher in primary cancer tissues than in adjacent noncancerous tissues, and in the proximal and distant margins of normal mucosa. Overexpression of Nup88 mRNA was statistically associated with TNM stage (P=0.044), lymphatic metastasis (P=0.022), and cancer location (P=0.036), while not related to gender, age of patients and histological type, infiltration depth, and differentiation of cancers. The expression of Nup88 mRNA in the HCT-116 p 53 − / − cell line was not significantly different from expression in the HCT-116 p 53 + / + cell line. And there was no correlation between Nup88 and p53 protein expression (r=0.632, P=0.368). Conclusions: Nup88 mRNA was overexpressed in colorectal cancers and the overexpression was associated with cancer development and aggressiveness. Nup88 might be regard as essential contributor to nodal metastagenicity of colorectal cancer. Show more

Keywords: Nup88, p53, mRNA, RT-PCR

DOI: 10.3233/DMA-2011-0842

Citation: Cancer Biomarkers, vol. 8, no. 2, pp. 73-80, 2011

Authors: Sarkar, Debansu | Singh, S.K. | Mandal, Arup K. | Agarwal, Mayank M. | Mete, Uttam K. | Kumar, Santosh | Mavuduru, Ravimohan S. | Prasad, Rajendra

Article Type: Research Article

Abstract: Objective: Development of castrate resistant prostate cancer (CRPC) indicates progressive disease with poor survival. Docetaxel has variable response with a good proportion of nonresponders. Neuroendocrine differentiation, hypothesised as one of the mechanisms behind development of CRPC, can be assessed by plasma Chromogranin A (CgA). We evaluated the clinical importance of circulating CgA in CRPC patients receiving Docetaxel. Methods: Plasma CgA was assessed by ELISA in 14 patients with CRPC receiving 3-weekly docetaxel. Baseline PSA, baseline CgA, PSA response, CgA response and clinical response to chemotherapy were evaluated and analysed. Results: Increased plasma CgA was observed in …64.3% of patients. There was no correlation between baseline CgA and PSA. Two patients with PSA < 10 ng/ml had elevated CgA. Baseline CgA was not influenced by variables such as site of metastasis and time to develop CRPC status. Seven patients (50%) had PSA-response and 5 (36%) CgA-response. In 4 patients PSA response and CgA response were discordant. Compared to men with normal baseline CgA, a higher proportion of those with elevated baseline CgA had PSA response (55% vs 40%), symptomatic response (66% vs 40%) and radiological response (55% vs 20%). Two patients with symptomatic response had only CgA response. Three patients having disease progression despite PSA response had increasing CgA. Conclusions: CgA and PSA are complementary tumour markers in CRPC. CgA may help in predicting the response of docetaxel therapy. Rising CgA during therapy may be associated with bad prognosis whereas CgA response is likely to be associated with clinical response. Show more

Keywords: Castrate resistant prostate cancer, docetaxel, Chromogranin-A, Neuroendocrine differentiation

DOI: 10.3233/CBM-2011-0198

Citation: Cancer Biomarkers, vol. 8, no. 2, pp. 81-87, 2011

Authors: Efrati, Edna | Elkin, Hela | Peerless, Yehudit | Sabo, Edmond | Ben-Izhak, Ofer | Hershkovitz, Dov

Article Type: Research Article

Abstract: Introduction: KRAS mutations in colon carcinomas are associated with lack of response to anti-EGFR monoclonal antibody treatment. Therefore, patients must undergo genetic testing to be eligible for treatment. Several methods for KRAS mutation analysis exist, but many are not sensitive enough to detect a mutation in samples with low fraction of malignant cells. In the present study, we developed a KRAS mutations detection method that is both simple and sensitive. Methods: Using a locked nucleic acid (LNA) containing oligonucleotide, we developed a PCR clamping method that preferentially amplifies the mutated over wild type KRAS. We evaluated the sensitivity …of this method using serial dilutions of plasmids containing wild-type and mutated KRAS fragments. Additionally, KRAS mutation status was evaluated on 60 archived tissue samples of colon carcinoma, and compared to direct sequencing and high resolution melting (HRM) methods. Results: The PCR clamping method could detect as little as 1% mutated DNA in the sample analyzed. Of the 29 KRAS mutations identified by the PCR clamping method, only 23 (79%) were identified by standard direct sequencing. The results of PCR clamping correlated with HRM results. Conclusions: LNA based PCR clamping method is a simple and highly sensitive method for the detection of KRAS mutations. Show more

Keywords: KRAS, colon carcinoma, locked nucleic acid (LNA), PCR clamp

DOI: 10.3233/CBM-2011-0203

Citation: Cancer Biomarkers, vol. 8, no. 2, pp. 89-94, 2011

Authors: Vidotto, Alessandra | Henrique, Tiago | Raposo, Luiz Sérgio | Maniglia, José Victor | Tajara, Eloiza H.

Article Type: Research Article

Abstract: Several body fluids have been evaluated as new sources for cancer biomarker discovery. In this context, salivary and serum proteomics seem promising diagnostic and predictive tools for head and neck diseases. In the present study, we performed a proteomic analysis of saliva and serum from patients presenting head and neck squamous cell carcinoma (HNSCC) and compared the results before and after therapy. In saliva of cancer patients, we observed an altered protein profile, including over-expression of PLUNC and zinc-alpha-2-glycoprotein. Both proteins may contribute to control tumor growth and, therefore, represent targets for new analysis. We also detected serotransferrin and a …modified transthyretin form with altered levels in serum from patients. Comparing preoperative and postreatment samples, the results showed that the protein profile after treatment reverted to a pattern closer to those observed for controls. These results add information on the role of secreted proteins in the cancer process and emphasize the potential of saliva and serum analysis for diagnosis and monitoring of HNSCC. Show more

Keywords: Head and neck carcinoma, saliva, serum, proteomics, radiotherapy

DOI: 10.3233/CBM-2011-0205

Citation: Cancer Biomarkers, vol. 8, no. 2, pp. 95-107, 2011

Authors: Ruffin, Mack T. | Normolle, Daniel P. | Evelegh, Michael J. | Baron, John A. | Bresalier, Robert S. | Marcon, Norman E. | Syngal, Sapna | Turgeon, D. Kim | Tuck, Melissa K. | Brenner, Dean E.

Article Type: Research Article

Abstract: The galactose oxidase-Schiff (GOS) reaction detects D-galactose-β-[1,3]-N-acetyl-D-galactosamine. This is a T-antigen expressed in mucus from malignant cells and colonic mucosa adjacent to cancer but not in normal mucosa. Previous studies using a qualitative GOS assay proved to be of limited value for the detection of colorectal neoplasia. We used a newly developed quantitative GOS assay to determine its potential as an early detection biomarker for colorectal cancer. We completed a multi-center, prospective, cross-sectional cohort validation study consisting of 70 normal controls, 23 high-risk normal patients (polyp history or family history of colorectal cancer (CRC) with currently normal colonoscopy), 137 patients …with adenomatous polyps, and 69 with colorectal cancers. Prior to colonoscopy, two samples of stool were collected via a rectal exam: one for FOBT, and one for GOS. The area under the ROC curve (AUC) for detecting colonic adenomas and cancer for normal colons, computed with logistic regression was 0.69 for GOS, 0.62 for FOBT, and 0.73 for GOS combined with FOBT. Adding GOS to FOBT did not significantly change the ROC of FOBT alone. GOS does not appear to be a suitable marker of colorectal neoplasia. Show more

Keywords: Colorectal Neoplasms/diagnosis, Colorectal Neoplasms/prevention & control, Carcinoma/diagnosis, Galactose Oxidase, Predictive Value of Tests

DOI: 10.3233/CBM-2011-0206

Citation: Cancer Biomarkers, vol. 8, no. 2, pp. 109-112, 2011