PIVKA-II as a Potential New Biomarker for Hepatocellular Carcinoma - A Pilot Study.

AIM: The aim of this study was to evaluate the clinical contribution of protein induced by vitamin K absence (PIVKA-II) for hepatocellular carcinoma (HCC) diagnostics and compare it with alpha-foetoprotein (AFP), a routinely used tumour marker. MATERIALS AND METHODS: A total of 332 participants were enrolled in this study: 64 with HCC, 48 with liver metastases of colorectal cancer origin, 42 with liver cirrhosis and 178 healthy individuals. Serum levels of PIVKA-II were measured using the chemiluminescent assay of the Architect 1000i System (Abbott, USA) and AFP levels using the chemiluminescent assay by DxI 800 (Beckman Coulter, USA). RESULTS: PIVKA-II achieved better clinical sensitivity than AFP and the difference in this sensitivity was statistically significant. PIVKA-II achieved the best sensitivity (96.9%) in distinguishing between the HCC and control groups with the proposed cut-off value of 60 mAU/ml. CONCLUSION: Our recommendation is for addition of PIVKA-II to the routine panel of HCC tumour markers.

CEA, CA 15-3, and TPS as Prognostic Factors in the Follow-up Monitoring of Patients After Radical Surgery for Breast Cancer.

AIM: The aim of this study was to evaluate the ability of tissue polypeptide-specific antigen (TPS), carcinoembryonic antigen (CEA), and cancer antigen 15-3 (CA 15-3) to predict relapse in breast cancer patients, when the measurement of biomarkers is performed within 6 months after surgery. PATIENTS AND METHODS: Four hundred and seventy-two patients with breast cancer were evaluated. TPS, CEA, and CA 15-3 were measured in months 1, 3, and 6, after surgery. Disease recurrence was recorded between 7-12 months after surgery. Disease recurrence occurred in 60 patients, while 412 patients remained in recurrence-free status. RESULTS: TPS levels of the recurrence group differed statistically significantly in the first and sixth month after surgery compared to recurrence-free group (p=0.0339, AUC=0.6056; p<0.0001, AUC=0.7196). CEA and CA 15-3 measurements did not achieve a statistically significant difference for any month examined. CONCLUSION: TPS level in the sixth month after surgery is the best candidate biomarker to predict disease recurrence.

Evaluation of IL-2, IL-6, IL-8 and IL-10 in Malignant Melanoma Diagnostics.

AIM: The aim of the present study was to evaluate the usefulness of four interleukins (IL-2, IL-6, IL-8 and IL-10) for melanoma detection and correlate these interleukins with sentinel node metastasis positivity. PATIENTS AND METHODS: A group of 236 persons was assessed: 175 patients with melanomas and 61 healthy persons. Melanoma patients were divided to four groups according to Breslow score. We determined IL-2, IL-6, IL-8 and IL-10 in each plasma sample. Interleukin plasma levels were assayed using a Human Cytokine Milliplex Map kit. Measurements were performed using the Bio-Plex MAGPIX Multiplex Reader. Plasma samples were collected prior to surgery or any other form of treatment. All melanoma diagnoses were histologically verified. RESULTS: We compared interleukin plasma levels in the healthy group and plasma levels in each Breslow score stage. In the first Breslow score stage, IL-2 (p<0.0001), IL-6 (p=0.0004) and IL-10 (p<0.0001) were positive. In the second Breslow score, stage IL-2 (p<0.0001), IL-6 (p<0.0001), IL-8 (p=0.0017) and IL-10 (p<0.0001) were positive. By comparing the group of positive and negative sentinel node metastasis, we observed a statistically significant difference in two interleukins: The median of IL-2 levels in the negative group was 5.88 pg/ml compared to 32.57 pg/ml in the positive group (p=0.0005). The median of IL-6 levels in the negative group was 4.80 pg/ml compared to 32.02 pg/ml in the positive group (p=0.0048). CONCLUSION: Interleukins IL-2, IL-6 and IL-10 are promising biomarkers of early-stage melanoma. IL-2 and IL-6 appear to be prognostic biomarkers.

Reference values of IGF1, IGFBP3 and IGF1/IGFBP3 ratio in adult population in the Czech Republic.

BACKGROUND: IGF1 is responsible for regulation of growth, metabolism and differentiation of human cells. IGFBP3 is the most abundant of the carrier proteins for IGF1 in the blood. IGF1/IGFBP3 molar ratio is an indicator of IGF1 bioavailability. We decided to create a file of reference ranges of IGF1, IGFBP3 and IGF1/IGFBPP3 ratio for the adult Czech population across the age spectrum. METHODS: We selected a group of 1022 subjects, 467 males and 555 females (ages 20-98 years), from several regions in the Czech Republic. The group consisted of blood donors and patients undergoing regular preventive examinations. Serum levels of IGF1 and IGFBP3 were measured using the following radioimmunoassay kits: IRMA IGF1 (Immunotech, Marseille, France) and IRMA IGFBP3 (Immunotech, Prague, Czech Republic). The IGF1/IGFBP3 ratio was also calculated. The following groups of patients were excluded: patients with diabetes, high blood glucose, high insulin levels, post-surgery patients, polymorbid patients, and subjects with oncological diseases. Subjects were divided into seven age-groups. Changes in the levels of observed analytes in each decade across the age spectrum were evaluated. All statistical analyses were performed by SAS 9.3 (Statistical Analysis Software release 9.3; SAS Institute Inc., Cary, NC, USA). RESULTS: All three parameters IGF1, IGFBP3 and IGF1/IGFBP3 decreased in parallel with decrease in age: p<0.0001, r=-0.64, -0.35 and -0.54, respectively. The dynamics of the decline was different between males and females. Linear regression models with age as independent variable fitted by gender are displayed in Fig. 1. Non-parametric reference interval curves (medians and 2.5th-97.5th percentiles) for IGF1, IGFBP3 and IGF1/IGFBP3 ratio as function of age by gender are displayed in Fig. 2(a,b,c). All medians and 2.5th-97.5th percentiles were plotted by cubic spline. For males, linear regression models were as follows: IGF1=291.34619-2.41211 × age, IGFBP3=2931.62778-6.11659 × age, IGF1/IGFBP3=0.02897-0.00021213 × age. For females, we plotted the following: IGF1=241.67406-1.98466 × age, IGFBP3=3688.60561-16.39560 × age, IGF1/IGFBP3=0.02029-0.00013233 × age. IGF1 was statistically significantly higher in males with p<0.0001 (Wilcoxon test) but decreased faster (p=0.0121). IGFBP3 was statistically significantly higher in females with p=0.0004 (Wilcoxon test) but decreased faster (p<0.0001). IGF1/IGFBP3 was statistically significantly higher in males with p<0.0001 (Wilcoxon test) but decreased faster (p<0.0001). CONCLUSION: Authors recommend using of a linear regression model based reference ranges for IGF1, IGFBP3 and IGF1/IGFBP3 ratio and using different reference ranges for genders.

Evaluation of IGF1 serum levels in malignant melanoma and healthy subjects.

BACKGROUND/AIM: There were two aims in the present study. The first was to evaluate the usefulness of insulin-like growth factor 1 (IGF1) for melanoma detection. The second was to correlate changes of serum levels of IGF1 with the Breslow score and sentinel node metastasis positivity. PATIENTS AND METHODS: We examined a group of 216 cases, 77 patients with melanomas and 139 healthy probands. We determined the serum IGF1 levels of each patient using an IRMA radioisotope IGF1 assay kit. Serum samples were collected prior to surgery or any other form of treatment. All melanoma diagnoses were histologically verified. RESULTS AND DISCUSSION: Based on the statistical evaluation between the melanoma group and group of healthy individuals, we observed statistically significant differences in IGF1 serum levels. The median IGF1 levels in the melanoma group was 154.1 ng/ml compared to 111.2 ng/ml in the group of healthy individuals (p=0.0036). The changes of the IGF1 levels related to the Breslow score categories were statistically significant (p=0.0027). Lastly, we compared the results between the positive and negative metastatic affection of the sentinel nodes. The median IGF1 levels in the negative group was 173.5 ng/ml compared to 205.8 ng/ml in the positive group. This difference was statistically significant (p=0.0407). CONCLUSION: Serum levels of IGF1 were significantly higher in patients diagnosed with melanoma compared to the healthy control group. The changes of the IGF1 levels related to the Breslow score categories were statistically significant. Serum levels of IGF1 were significantly higher in the group with the positive metastatic affection of the sentinel nodes than in negative patients.

CA125 and HE4 levels in a Czech female population diagnosed with endometrial cancer in preoperative management.

AIM: The aim of the present study was to compare the use of cancer antigen 125 (CA125) and human epididymis protein 4 (HE4) biomarkers in patients with endometrial cancer for preoperative management and to particularly focus on relationship between CA125 and HE4 and disease stage in predicting myometrial invasion or intrauterine tumor spread. PATIENTS AND METHODS: Thirty-four patients diagnosed with endometrial cancer and 32 healthy controls were enrolled into the pilot study in the period between May 2012 and March 2013. Blood from all the females was collected and examined for CA125 and HE4. Based on standardized ultrasound examination, including gynecological examination, the clinical disease stage was determined. RESULTS: We found a significant difference (p<0.0001) for means of serum levels of HE4: females with endometrial cancer, 75.5 pmol/l, versus healthy females, 40.0 pmol/l. A non-significant statistical difference was found for mean serum CA125 levels (p=0.4442): females with endometrial cancer 19.0 IU/l, versus healthy females, 15 IU/l. A significant correlation with histopathological disease stage was found for both biomarkers (Spearman correlation). Sensitivity and specificity, and the related cut-off for HE4 suggest that HE4 would be a more appropriate biomarker for differential diagnosis between benign and malignant states. CONCLUSION: Based on our pilot study, we found that parallel examination of HE4 and CA125 may support endometrial ultrasound finding verification prior to biopsy. This study is ongoing and we expect that results on a larger population may enable HE4 measurement to be implemented in routine practice.

Conference scene: ISOBM 2013 in Israel: part 1: biomarkers - where we are now?

The 40th annual Congress of the International Society of Oncology and Biomarkers took place in Jerusalem, Israel. The subtitle of this year's conference was: 'Innovations in oncological treatments and tumor markers'. Many issues on the potential use of markers were discussed, such as the potential use of markers for therapy response prediction in personalized medicine and for the proposal of algorithms for clinical practice use. A series of lectures was presented related to the discovery and preclinical and clinical validation of new biomarkers. The presentations were also split into keynote lectures, oral presentation in sessions and poster presentations. The conference was attended by approximately 186 participants and was attended by some of today's leading laboratory researchers in the field of oncology research. When comparing this conference with other meetings from previous years, it was evident that the biomarkers that are gaining in significance are DNA changes in association with the cancer process and epigenetic changes, which is seen to be methodically moving the field of immunoassays into molecular biology methods.

Conference scene: ISOBM 2013 in Israel: part 2: biomarkers - a challenging issue for futher investigation.

This is the second article dedicated to The Annual 40th Congress of the International Society of Oncology and Biomarkers, which took place in Jerusalem, Israel. This second part of this article will be dedicated to the special presentations from individual sessions. Sessions were organised according to the cancer type or etiopathogenesis, while some sessions were dedicated to the relationship between diagnostics and therapy choice.

Growth factors and breast tumors, comparison of selected growth factors with traditional tumor markers.

BACKGROUND: The first aim of this project was to study new possibilities for distinguishing benign from malignant tumors using growth factors and to compare them with the traditional tumor markers Carcinoembryonic antigen (CEA) and Cancer antigen 15-3 (CA15-3) for breast tumors. The second aim was to make a comparison of CEA, CA 15-3, Insulin-like growth factor I (IGF1), Insulin-like growth factor-binding protein 3 (IGFBP3), Hepatocyte growth factor (HGF) and Epidermal growth factor (EGF) for individual stages of cancer. PATIENTS AND METHODS: Our group of patients consisted of 110 females, 89 with breast cancer and 21 with benign breast tumors (fibroadenomas). Serum levels of CEA and CA 15-3 were measured using a DxI instrument. Serum levels of IGF1 and IGFBP3 were measured using IRMA radioisotope assay kits. HGF and EGF were measured using an xMAP Luminex multiplex panel. Serum samples were collected prior to surgery and those of the two groups of patients were compared (malign vs. benign). Patients with diabetes mellitus were excluded from this project. RESULTS AND DISCUSSION: Comparing the individual parameters of serum levels between the two groups of patients (malignant vs. benign) only HGF was found to show a statistically significant difference. The mean of HGF in patients with malignant diseases prior to surgery was 3370 pg/ml compared to 1799 pg/ml in benign tumors with p=0.0016. We found significantly lower serum values of IGF1 at stage III in comparison to stages I and II: mean values: at stage I=181 ng/ml, at stage II=182 ng/ml and at stage III=70 ng/ml; stage III vs. stage II, p=0.0167. CONCLUSION: Tumor markers are currently used for therapy monitoring in cancer patients as one of the indicators of successful therapy. Our findings correspond to existing literature. IGF1 and its binding protein IGFBP3 cannot be used to distinguish between malignant and benign tumor. HGF is considered to be a marker of progression and of the aggressiveness of breast cancer; our data fully corresponds to this. Based on our data, this marker could potentially be used as an additional tool for the differentiation between benign and malignant tumor.

Vitamin D in colorectal, breast, prostate and lung cancer: a pilot study.

BACKGROUND: Many studies have demonstrated the relationship between vitamin D and cancer of many different sites, including of the breast, colorectum, prostate and lung. Most epidemiological studies have assessed the effects of dietary intake only, although endogenous production after sun exposure is the main source of vitamin D. The aim of our pilot study was to study serum levels of vitamin D in general population and in patients with different type of cancer. PATIENTS AND METHODS: The control group consisted of 214 healthy individuals. Pathological groups of patients included 170 patients with different cancer types (28 patients with prostate cancer, 43 patients with breast cancer, 49 patients with colorectal cancer and 50 patients with lung cancer). All of the patients were enrolled in the early clinical stage of cancer up to clinical stage III. Advanced stages were not included into the study. Vitamin D serum levels were measured using ECLIA Roche method. RESULTS: All the results for serum vitamin D from pathological groups were significantly lower compared to the levels of the control group. All the cancer types had a high incidence rate of very low serum levels of vitamin D. Lung cancer had the highest incidence rate of very low vitamin D serum levels. CONCLUSION: We found a high incidence of hypovitaminosis D in cancer patients compared to a healthy control group among a Czech population. This incidence rate is higher in comparison to data found in literature from the other parts of the world. Based on the data from this study, a large epidemiological study monitoring vitamin D serum levels in the healthy population and in cancer patients in the Czech Republic has been already proposed.

The role of MonoTotal in the primary diagnosis, prognosis and follow-up of patients with non-small cell lung cancer (NSCLC).

BACKGROUND: A new cytokeratin tumor marker, MonoTotal was studied in lung cancer, the most common cause of cancer mortality worldwide. In non-small cell lung cancer (NSCLC) patients MonoTotal serum levels and their relationship to the tumor stage, histological subtype, early relapse and cancer-related death were evaluated. PATIENTS AND METHODS: MonoTotal serum levels were studied, using immunoradiometric assay in a group of 93 patients with newly diagnosed NSCLC undergoing radical surgery, and were compared to those with benign lung diseases. RESULTS: A diagnostic power of MonoTotal in distinguishing patients with NSCLC from benign lung diseases was demonstrated. Higher levels of MonoTotal were associated with advanced stages of squamous cell carcinoma and there was a positive correlation of marker with tumor size. Marker levels showed significant relation to disease-free survival and overall survival. CONCLUSION: MonoTotal seems to be a potentially very interesting serum marker that, in conjunction with other clinical data, might be used for monitoring of patients with NSCLC.

Parameters of biological activity in colorectal cancer.

BACKGROUND: The aim of this study was to measure several parameters in patients with early-stage colorectal cancer (CRC) and to evaluate them for their utility in routine clinical practice. PATIENTS AND METHODS: Pre-operative serum levels of the following parameters were measured in 174 patients with CRC (clinical stage I-III): carcinoembryonic antigen (CEA), carbohydrate antigen CA 19-9, proliferative marker thymidine kinase (TK), tissue polypeptide antigen (TPA), tissue polypeptide-specific antigen (TPS), interleukin-6 (IL-6), interleukin-10 (IL-10), matrix metalloproteinase-9 (MMP-9), tissue inhibitor of metalloproteinase-1 (TIMP-1), intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), vascular endothelial growth factor (VEGF), C-peptide, insulin, adiponectin and leptin. The control group consisted of 50 patients who were undergoing a complete preventive medical examination and in these patients at the time of blood collection there was no evidence of any cancer disease. RESULTS: Significant increase of the following parameters was found in patients with CRC: CEA, CA 19-9, TPA, IL-6, IL-10, TIMP-1, C-peptide, insulin and adiponectin. Only two of these, CA 19-9 and adiponectin, represent highly unfavorable prognostic factors. If elevated, they affect both progression-free interval and overall survival. CONCLUSION: Based on our results, we can conclude that none of the measured parameters fulfills the criteria for use for screening nor for primary diagnosis of CRC. Some of the parameters are important for prognosis estimate: Elevated CA 19-9 is related to an unfavorable prognosis, in terms of cancer recurrence and mortality rate. Angiogenetic factor VEGF represents a prognostic factor important for OS. CEA represents a parameter which is related to disease progression. Interleukins seem to be prospective complementary tumor markers. Adiponectin may be used for estimation of advanced stage of cancer and for estimate of risk of cancer recurrence.

Tumor markers in patients with relapse of colorectal carcinoma.

AIM: To evaluate CEA and CA19-9 in a long-term follow-up after radical surgery for colorectal cancer. PATIENTS AND METHODS: A total of 1,090 patients were operated on for colorectal cancer, 716 patients underwent R0 resection, 631 patients were under further surveillance, relapse was diagnosed in 122 patients (20%), 74 patients were indicated for reoperation The resectability of the relapse was 35%. An AxSYM instrument (Abbott) was used for analysis. RESULTS: At the time of relapse both markers were normal in 31% of the patients. When relapse was diagnosed, in patients with normal preoperative levels, CEA and CA19-9 were below cut-off in 48% and 79%, respectively, and in those with primary elevation, they were again elevated in 78% and 64%, respectively. CONCLUSION: The surveillance based only on CEA and/or CA19-9 was cost-effective, but failed to disclose 1/3 of patients suffering from relapse; these markers must be combined with liver and chest imaging methods and colonoscopy.

Multiorgan resections for advanced colorectal cancer.

BACKGROUND: A retrospective review is presented of a single institution's experience with multivisceral resections for locally-advanced colorectal cancer. MATERIALS AND METHODS: Twenty-eight patients, who had undergone RO multiorgan resection, were identified from the database of a total of 1150 patients operated on for colorectal carcinoma in the years 1995-2005 at a single center. There were twelve total pelvic exenterations and 16 patients had undergone en bloc primary tumor resection with adherent organs, such as the spleen, diaphragm, pancreas, stomach, kidney, etc. The patients were followed-up according to a standard protocol. RESULTS: The post-operative mortality was 7%, the average follow-up 21.6 months and the 5-year survival 45%. CONCLUSION: Our results confirmed that, in the case of invasion of colorectal cancer to the adjacent intra-abdominal organs or structures, multiorgan resection 'offers the only chance of potentially-curative treatment.