The significance of interleukin-8 in hormonally dependent early breast cancer – association with the established parameters ER/PR and HER2

B ackground: Interleukin-8 (IL-8) is a mulƟ funcƟ onal cytokine linked to cancer progression. Studies have confi rmed high IL-8 levels in HER2-enriched and basal-like (ER–) primary breast tumors. The aim of this study was to evaluate the relaƟ onship between intratumoral IL-8 protein levels and clinical outcome in hormone dependent (ER+) primary breast cancer paƟ ents. PaƟ ents and methods: The study included 65 early-stage breast cancer paƟ ents with detectable levels of hormone receptors (ER>0, PR>0), all of whom had not received any prior hormonal or chemotherapeuƟ c systemic therapy. The median follow-up was 144 months. Steroid hormone receptor status was determined by ligand-binding assay. HER2 status (absence or presence of gene amplifi caƟ on) was determined by chromogenic in situ hybridizaƟ on (CISH). IL-8 protein levels were determined in cytosol tumor extracts by quanƟ taƟ ve ELISA. ER level of 10 fmol/mg, PR level of 20 fmol/mg and the median IL-8 concentraƟ on level of 88.8 pg/mg, were used as cut-off values. Results: There was a signifi cant diff erence in relapse free survival (RFS) between IL8low and IL8high subgroups of paƟ ents (Log rank test, p=0.002). Considering subgroups of paƟ ents straƟ fi ed in diff erent phenotypes according to receptor status and the median IL-8 value, if IL-8 is highly expressed, the infl uence of ER is weaker and there was no signifi cant diff erence in RFS between subgroups with ERlowIL8high and ERhighIL8high phenotypes. The same is true for PR and HER2 and there was no signifi cant diff erence in RFS between subgroups with PRlowIL8high and PRhighIL8high phenotypes, neither between subgroups with HER2−IL8high and HER2+IL8high phenotypes. On the other hand, subgroup with ERhighIL8low phenotype had signifi cantly longer RFS compared to those with ERlowIL8high and ERhighIL8high phenotypes (p=0.02, p=0.04, respecƟ vely); subgroup with PRlowIL8low phenotype had signifi cantly longer RFS compared to those with PRlowIL8high and PRhighIL8high phenotypes (p=0.003, p=0.02, respecƟ vely); and subgroup with HER2–IL8low phenotype had signifi cantly longer RFS compared to those with HER2–IL8high and HER2+IL8high phenotypes (p=0.01, p=0.02, respecƟ vely). Conclusions: IL-8 is a potenƟ al biomarker of unfavorable prognosis in hormone dependent breast cancer that is associated with the established parameters ER/PR and HER2. Receptor-mediated signaling could act addiƟ vely with IL-8 signaling in progression of hormone dependent breast cancerThe first number of Oncology Insights includes Proceedings book of The Sixth Congress of the Serbian Association for Cancer Research with international participation (Oct 2-4, 2023, Belgrade

The prognostic significance of serum interferon-gamma (IFN-γ) in hormonally dependent breast cancer

Background: Interferon-γ (IFN-γ) is a pleiotropic immunomodulatory cytokine. Because of its contradictory and even dualistic roles in malignancies, its potential as a biomarker remains to be unraveled. Aim: To evaluate the prognostic significance of serum IFN-γ in hormonally treated breast cancer patients. Material and methods: The study included 72 premenopausal breast cancer patients with known clinicopathological characteristics. All patients received adjuvant hormonal therapy based on hormone receptor-positivity. The median follow-up period was 93 months. IFN-γ serum protein levels were determined by quantitative ELISA. Prognostic performance was evaluated by the receiver operating characteristic (ROC), Cox proportional hazards regression and Kaplan-Meier analyses. Classification of patients into IFN-γlow and IFN-γhigh subgroups was performed by the use of the outcome-oriented cut-off point categorization approach. Results: The best prognostic performance was achieved by IFN-γ (AUC = 0.24 and p = 0.01 for distant events, AUC = 0.29 and p = 0.01 for local and distant events combined). Age and IFN-γ were prognostically significant in instances of all types of outcomes and IFN-γ was the independent prognostic parameter (Cox regression). There was a significant difference between IFN-γ values of patients without any events and those with distant metastases (Mann-Whitney test, p = 0.007). IFN-γ levels correlated significantly with nodal status and tumor stage (Spearman's rank order, r = −0.283 and r = −0.238, respectively). Distant recurrence incidence was 4% for the IFN-γhigh subgroup and 33% for the IFN-γlow subgroup (Kaplan–Meier analysis). Conclusions: Raised serum IFN-γ levels associate independently with favorable disease outcome in hormonally dependent breast cancer

The significance of HOXB7 and IL17RB serum levels in prognosis of hormonally dependent breast cancer: A pilot study

Purpose: Improved prognostication of a patient's outcome could allow for personalized treatment decisions in breast cancer. Homeobox B7 (HOXB7) and interleukin 17 receptor B (IL17RB) are proteins reportedly involved in the development of hormonal therapy resistance. Their prognostic value was previously investigated in tumor tissue but recent mass spectrometric detection of HOXB7 and IL17RB proteins in serum has prompted us to perform the first prognostic evaluation of their serum levels. Patients and methods: The study included 81 premenopausal breast cancer patients that received adjuvant hormonal therapy. The median follow-up period was 61 months. HOXB7 and IL17RB serum protein levels were measured by quantitative sandwich ELISA and prognostically evaluated by Cox proportional hazards regression analysis. Results: HOXB7 protein was detected in 96.3% and IL17RB in 33.3% of serum samples. Higher levels of serum HOXB7 significantly associated with favorable disease outcome by prognosticating distant (by HR ​= ​0.04; P ​= ​0.001) and local recurrence (by HR ​= ​0.03, P ​= ​0.001). The recurrence rates in the HOXB7high and HOXB7low subgroups of patients (cut-off 81.5 ​pg/mL) were 0% and 17%, respectively. Serum IL17RB levels did not significantly associate with either local or distant events. The multivariate analysis highlighted estrogen receptor, histological grade, nodal status and HOXB7 as independent prognostic parameters. Conclusions: Our findings validate the previous mass-spectrometry data by showing that HOXB7 and IL17RB cellular proteins are detectable in serum by a standard ELISA assay. Furthermore, we show that HOXB7 serum levels are the relevant prognosticator of response to hormonal therapy

The importance of simultaneous determination of breast cancer biomarkers

It is shown that steroid hormone receptors by themselves are not sufficiently strong prognostic factors in management of breast cancer. For that reason, simultaneous consideration of different biomarkers seems to be more appropriate for clinical use, i.e. selections of patients with high/inter-mediate/low risk of disease outcome. However, the amount of tumor material available from breast carcinoma can preclude determination of estrogen- regulated biomarkers together with estrogen receptor and progesterone receptor. The aim of this study was to assess the possibility of estrogen receptor and progesterone receptor determination by a single-point instead of five-point biochemical method. Our results demonstrated that the correlation between measurements of estrogen and progesterone receptor contents obtained by the five-point and single-point assay in the total population was very high. Consequently, we could use the single-point assay instead of five-point assay for estrogen receptor and progesterone receptor determination, thus making possible determination of other molecular biomarkers from the same breast carcinoma

Can granulysin provide prognostic value in primary breast cancer?

Granulysin (GNLY) is a cytolytic and proinflammatory molecule which also acts as an immune alarmin. The multifunctional nature of this molecule has made it challenging to define its full potential as a biomarker in breast cancer.AimTo evaluate the prognostic value of intratumoral GNLY in primary breast cancer patients and its association with established clinicopathological parameters.Patients and methodsThe study included 69 node-negative breast cancer patients with known clinicopathological parameters, all of whom had not received any prior hormonal or chemotherapeutic systemic therapy that would interfere with the course of disease. The median follow-up period was 144 months. Steroid hormone receptor status was determined by ligand-binding assay and HER2 status by chromogenic in situ hybridisation (CISH). Intratumoral GNLY mRNA levels were determined by RT-qPCR. Prognostic performance was evaluated by the receiver operating characteristic (ROC), Cox proportional hazards regression and Kaplan-Meier analysis. Classification of patients into GNLYlow and GNLYhigh subgroups was performed by the use of the outcome-oriented cut-off point categorisation approach.ResultsThere was a significant difference between GNLY values of patients without any recurrences and those with local or distant recurrences (Mann-Whitney test, p = 0.05 and p = 0.02, respectively). None of the tested parameters showed prognostic significance for local and distant recurrences when combined. When distant metastases and local recurrences were separated as events, the best prognostic performance was observed for GNLY as compared with any clinicopathological parameter (AUC=0.24 and p = 0.04 for local events; AUC=0.71 and p = 0.03 for distant events). Local recurrence incidence was 0% for the GNLYhigh subgroup and 19% for the GNLYlow subgroup; however distant recurrence incidence was 24% for the GNLYhigh subgroup but only 3% for the GNLYlow subgroup (Kaplan–Meier analysis). A significant positive correlation was found between intratumoral ER and GNLY levels, and a significant negative correlation between tumour grade and GNLY levels.ConclusionHigh levels of granulysin prognosticate low risk of local recurrence but a high risk of distant metastasis in primary, untreated, breast cancer patients

Vascular endothelial growth factor (VEGF) -A, -C and VE-cadherin as potential biomarkers in early breast cancer patients

Background: Vascular endothelial growth factor (VEGF) -A and -C act as multifunctional molecules and growth factors, while VE-cadherin (cadherin 5, CDH5) is the endothelial junction protein. Aim: To assess the relationship between intratumoral VEGF -A, -C and CDH5 levels and clinical outcome, in primary, early-stage, breast cancer patients. Patients and methods: The study included 69 node-negative (N0) breast cancer patients, all of whom had not received any prior hormonal or chemotherapeutic systemic therapy that would affect the course of disease. The median follow-up period was 144 months. Intratumoral mRNA levels of VEGF -A, -C and CDH5 were determined by RT-qPCR. Prognostic performance was evaluated by Cox proportional hazards regression, Kaplan-Meier analysis, as well as by the multivariable approach based on the least absolute shrinkage and selection operator (LASSO) logit regression. Classification of patients into the low and high subgroups was performed using the outcome-oriented cut-off point categorization approach. Results: Of the measured mRNAs, only CDH5 mRNA (t = −2.17; p = 0.04) and VEGF-C mRNA (t = −2.41; p = 0.03) showed significant differences between values in patient subgroups with distant metastasis and those without recurrences, respectively. These t-test results were in agreement with the Cox regression by which CDH5 mRNA reached the most pronounced hazard ratio (HR=2.07; p = 0.05), followed by VEGF-C mRNA (HR=1.59; p = 0.005). HR values above 1.0 indicate that high levels of either CDH5 or VEGF-C mRNAs associated with a higher risk of poor clinical outcome. Distant recurrence incidence was 26% for the CDH5high and 3% for the CDH5low subgroup (Kaplan–Meier analysis). Distant recurrence incidence was 23% for the VEGF-Chigh and 0% for VEGF-Clow subgroup. The independent prognostic value of VEGF-C mRNA was confirmed by LASSO regression. Conclusion: Intratumoral VEGF-A levels did not associate with disease outcome in primary, early-stage, breast cancer patients, whilst raised levels of either CDH5 or VEGF-C prognosticated a high risk of distant metastasis

TGF-beta1 in breast cancer-estrogen regulation

TGF-beta1 is a pluripotent cytokine with diverse effects in the normal development of mammary glands, and in the development of malignant tumors of the breast. The aim of the study was to determine the levels of TGF-beta1 in the group of advanced breast cancer, in which increased TGF-beta1 levels were most likely to be expected. TGF-beta1 levels were also compared with estradiol levels. Our results suggested that TGF-beta1 synthesis may be regulated by estrogen or anti-estrogen through ER. Finding of increased TGF-beta1 levels, due to its possible role in predicting invasive phenotype in later phases of tumor progression, may indicate the tendency of tumor tissue towards autonomy