The Effects of Oleuropein on Different Clinically Types of Human Neuroblastoma Cells

Neuroblastoma is an embryonic tumor originating from the neural crest. It accounts for 8–10% of all childhood cancers. Although Cisplatin is used in neuroblastoma treatment, it has many side effects, such as ototoxicity, nephrotoxicity, and neurotoxicity. One herbal agent that has attracted attention in recent years is oleuropein (OLE), the active component of olive leaf. This component belongs to the polyphenol group and it has antioxidant, anti-microbial, anti-inflammatory, anti-hypertensive and anti-carcinogenic effects. It has beneficial effects against neurodegeneration in both culture cells and model organisms. Oleuropein has been shown to be increased apoptosis in SH-SY5Y neuroblastoma cell line in one study. Cisplatin (cis-diaminedichloroplatinum II (CDDP) is a widely used agent for the treatment of many different human cancers in childhood and adults with antimitotic and antineoplastic properties. CDDP is the most effective chemotherapeutic agent in specially treatment of neuroblastoma. Purpose of this study was to determine whether oleuropein and CDDP have possible anti-proliferative activity in different types of human neuroblastoma cells as representing different clinical features (bone marrow metastatic LAN-5 cells and treated with chemotherapy and beam therapy CHP-134 cells representing late relapse) investigated. Human bone marrow metastatic LAN-5 and treated with chemotherapy and beam therapy CHP-134 neuroblastoma cells representing late relapse were used in this study. The effects of OLE and CDDP on LAN-5 and CHP-134 neuroblastoma cell proliferation and apoptotic cell death was investigated using WST-1 cell proliferation and Annexin-V/PI flow cytometric assays. Oleuropein and CDDP have been shown to inhibit proliferation of LAN-5 and CHP-134 neuroblastoma cells. In further studies, it is planned to investigate different cell death mechanisms by using combination of oleuropein and cisplatin in different kind of human neuroblastoma cells

“Don’t Eat Me” Signals of Neuroblastoma by CD47 for Immune Escape: A Novel Prognostic Biomarker

Recent studies have shown that cancer cells can deceive phagocytosing macrophage cells through the CD47 protein which gives the message “don’t eat me” or “don’t kill me” to immune components. The efficacy of anti-CD47 treatment approach was shown in cancers such as, non-small cell lung cancer, non-Hodgkin lymphoma, ovarian cancer, and breast cancer. The studies on the immunobiology of neuroblastoma has increased as monoclonal antibody based immunotherapy has shown to be effective in high-risk patients such as anti disialoganglioside. Therefore, the aim of this study was to evaluate the levels of CD47 protein expression among neuroblastoma patients with different risk groups and genetic alterations. This study included paraffin-embedded tumor tissues of 66 neuroblastoma patients (28 girls, 38 boys) with an age range of 0.5 to 108 months with a mean value of 24.9 (±23.5). According to risk classifications 21 were at low risk (31, 8%), 24 were at intermediate risk (36, 4%) and 19 were at high-risk (28, 8%) groups. These samples were evaluated for MYCN amplification, 1p36 LOH, 11q23 deletion and 17q25 gain by real-time PCR. In addition, CD47 expression status (positive or negative) was detected by immunohistochemical analysis. All data was analyzed with Chi-Square and Mann-Whitney U non-parametric tests within SPSS program, version 22.0. p value lower than 0, 5 was found statistically significant. According to the results, patients at low risk did not express CD47, while patients at high-risk group were mostly expressing CD47 (p = 0.049). MYCN amplification positive patients were expressing CD47 protein (p = 0.046). Patients without 17q25 gain were found to be expressing CD47 protein (p = 0.006). In addition, CD47 expression was increasing as age was getting higher in terms of months (p = 0.018). The findings of this study suggest that positive expression pattern of CD47 may be a poor prognostic biomarker especially in high risk 17q gain negative neuroblastoma patients

Molecular Heterogeneity in Neuroblastoma and Its Clinical Significance

Objective: Tumor heterogeneity describes the differences between cancer cells in the same tumor sample. Neuroblastoma (NB) is a type of cancer where tumor heterogeneity complicates its treatment. This study aims to explore the role of molecular heterogeneity detected by routine molecular tests in NB. Method: Seventy-one patients were included in the study. NB samples were chosen among 1,300 NB samples that were evaluated using molecular tests between 2012-2020 according to the guidelines of Turkish Pediatric Oncology Group Protocol. Molecular investigations were performed (total 142 samples) obtained from two different areas of the tumor (synchronous) or at two different times (metachronous). Heterogeneity was questioned for five tests: MYCN amplification, 1p36LOH, 11q2 3 deletion and 17q25 gain (identified with real-time polymerase chain reaction) and DNA ploidy (identified with flow cytometry). Results: Heterogeneity was observed for MYCN in 22.53%, for 1p36LOH in 36.62%, for 11q23del in 29.58%, and for 17q25 gain in 40.85% of cases, while DNA ploidy was heterogeneous in 36.4% of cases. Molecular heterogeneity did not show statistical difference among metachronous and synchronous cases. High-risk cases more frequently displayed molecular heterogeneity without any statistically significant difference between both groups. Conclusions: Our findings support the fact that molecular heterogeneity either exists in different areas of a tumor or seen in the same tumor at different times. It will be beneficial to perform more than one molecular analysis on the tumor tissue specimens. In addition, recurrences or re-biopsy specimens from metachronous metastases shall be re-evaluated using molecular tests in cases of NB