Glioblastoma multiform (GBM) is one of the most severe tumor types. It is highly invasive and characterized as a grade IV neoplastic cancer. Its resistance to chemotherapy-temozolomide (TMZ treatment)-in combination with tumor treating fields (TTFields), limits the cure of GBM. Therefore researchers are searching for new treatment options to increase the length of recurrence time and improve overall survival for GBM patients. Several cell lines have been established and are in use to understand the molecular basis of GBM and to test the developed drugs. On one hand, it is highly advantageous to utilize multiple cell lines with different genetic backgrounds to gain more insight into the characterization and treatment of the disease. However, on the other hand, characteristics of these cell lines such as proliferation rate, invasion, and colony formation capacity differ greatly among these cells. Hence, a detailed comparison concerning molecular and cellular features of commonly used cell lines is essential. In this study, cell proliferation and apoptosis rate, cell migration capacity, and gene expression profile of U87, Ln229, and SvGp12 cells have been investigated and compared.
The regenerative capacity varies significantly among the animal kingdom. Successful regeneration program in some animals results in the functional restoration of tissues and lost structures. Among the highly regenerative animals, axolotl provides multiple experimental advantages with its many extraordinary characteristics. It has been positioned as a regeneration model organism due to its exceptional renewal capacity, including the internal organs, central nervous system, and appendages, in a scar-free manner. In addition to this unique regeneration ability, the observed low cancer incidence, its resistance to carcinogens, and the reversing effect of its cell extract on neoplasms strongly suggest its usability in cancer research. Axolotl's longevity and efficient utilization of several anti-aging mechanisms underline its potential to be employed in aging studies.
Background: Grade IV neoplasm of the central nervous system, GBM, is associated with poor prognosis and short overall survival. Objectives: This study aimed to investigate the effects of conditioned mediums of GBM cell lines on each other. Methods: Conditioned mediums of GBM cell lines were harvested at the 6th, 12th, 24th, and 48th h time points. The cellular and molecular effects of conditioned mediums were evaluated using gold standard techniques such as MTT assay, colony formation assay, wound healing assay, EdU labeling-based flow cytometry, and qRT-PCR. Results: Our study demonstrated that conditioned medium harvested from U87 or LN229 cells at the 48th h time point exhibited an anti-growth activity on each other by altering the gene expression pattern. Furthermore, the conditioned medium of LN229 decreased the migration capacity of U87 cells, and the conditioned medium of U87 cells significantly suppressed the LN229 proliferation. Conclusions: Growth-limiting activity achieved by conditioned mediums collected at the 48th h time point positioned them as promising candidates to further investigate their content. It was argued that this initial work provided new insights to expand our current understanding of the roles of GBM cells' secretome.
Breast invasive cancer (BIC) is one of the most commonly observed and the deadliest cancer among women. Despite the progress that has been made in improving breast cancer outcomes by the development of advanced treatment options, due to the heterogeneity and complexity of the disease, more studies are required to explore underlying molecular mechanisms of breast cancer which may provide useful insights to overcome the constraints related to current therapeutic options. The goal of this study was to reveal the crucial roles of m6A regulatory proteins in BIC development using various publicly available datasets and databases. We first conducted a comprehensive analysis to depict the mutation frequency and types for m6A regulatory genes in sub-types of BIC for the evaluation of the genetic alterations landscape of breast cancer. Changes in expression levels of m6A regulatory factors were identified as the key genetic alteration in BIC. Implementation of Kaplan-Meier tool to assess the predictive value of m6A pathway components in BIC validated the use of VIRMA, METTL14, RBM15B, EIF3B, YTHDF1, and YTHDF3 as prognostic biomarkers of breast cancer. We then examined the enriched gene ontology (GO) terms and KEGG pathways for the tumor samples with genetic alterations in the m6A pathway. Dysregulation of m6A regulatory factors in BIC was associated with cell division and survival-related pathways such as nuclear division, and chromosome segregation via the upregulation of the genes functioning in these biological processes, and the gained overactivity of these pathways may account for poor prognosis of the disease. The performed analyses highlighted m6A pathway genes as potential regulators of BIC growth and as a valuable set to be utilized as clinical biomarkers in BIC disease.
Grade IV neoplasm of the central nervous system, GBM, is associated with poor prognosis and relatively short overall survival. Due to the current limitations in treatment methods, GBM is characterized as an incurable disease, and research to advance therapeutic options is required. Conditioned medium is commonly used in in-vitro studies complementary to animal experiments to simulate tumor microenvironment and has the potential to challenge and expand our current understanding of secretome effect on tumor characteristics. This study aimed to investigate the effects of conditioned mediums of GBM cell lines on each other. Conditioned mediums' cellular and molecular effects were evaluated using commonly employed techniques such as MTT assay, colony formation assay, wound healing assay, EdU labeling-based flow cytometry, and qRT-PCR. Our study demonstrated that conditioned medium harvested from U87 or LN229 cells at 48th h exhibited an anti-growth activity on each other by changing the gene expression pattern. Furthermore, the conditioned medium of LN229 decreased the migration capacity of U87 cells, and the conditioned medium of U87 cells significantly suppressed the LN229 proliferation. We believe that this initial work provides new insights for a better understanding of GBM cell lines' secretome roles and highlights the necessity of further studies to unveil the secretome content.
Background: Glioblastoma multiforme (GBM) is a lethal form of central nervous system cancer with a lack of efficient therapy options. Aggressiveness and invasiveness of the GBM result in poor prognosis and low overall survival. Therefore, the necessity to develop new anti-carcinogenic agents in GBM treatment is still a priority for researchers. Ion channels are one of the primary regulators of physiological homeostasis with additional critical roles in many essential biological processes related to cancer, such as invasion and metastasis. A multi-channel blocker, hydroquinidine (HQ), is currently in use to treat short-QT and Brugada arrhythmia syndromes. Objective: The objective of the study was to examine the alterations in survival, clonogenicity, migration, tumorigenicity, proliferation, apoptosis, and gene expression profile of temozolomide (TMZ)-sensitive and TMZ-resistant GBM cells upon HQ treatment. Methods: The possible anti-neoplastic activity of HQ on GBM cells was investigated by several widely applied cell culture methods. The IC50 values were determined using the MTT assay. Upon HQ treatment, the clonogenicity and migration capacity of cells were evaluated via colony-formation and wound healing assay, respectively. For anti-proliferative and apoptotic effects, EdU and CFSE, and Annexin-V labeling were applied. Tumorigenicity level was depicted by employing soft agar assay. The expression level of multiple genes functioning in the cell cycle and apoptosis-related processes was checked utilizing qPCR. Results: A significant anti-carcinogenic effect of HQ on TMZ-sensitive and -resistant GBM cells characterized by the increased apoptosis and decreased proliferation rate was revealed due to the altered gene expression profile related to cell cycle and cell death. Conclusion: In this study, the anti-carcinogenic effect of HQ has been demonstrated for the first time. Our data suggest the possible utilization of HQ to suppress the growth of GBM cells. Further studies on GBM-bearing animal models are required to assess its therapeutic potential in GBM treatment.
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