Cancer
Permanent URI for this collectionhttps://hdl.handle.net/20.500.12503/32539
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Browsing Cancer by Author "Basha, Riyaz"
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Item EXPLORING A COMBINATION USING CHEMOTHERAPY AND TOLFENAMIC ACID TO INDUCE ANTI-PROLIFERATIVE RESPONSE IN MEDULLOBLASTOMA CELL LINES(2024-03-21) Chen, Liling; Sankpal, Umesh; Basha, RiyazBackground: Medulloblastoma (MB) is the most common malignant brain tumor in children, with a peak incidence between the ages of 5-9 years old. Originating in the cerebellum, it often metastasizes throughout the CNS via the CSF, making it very difficult to treat. Current treatment options are limited, and includes surgical resection followed by radiation and chemotherapeutic agents. However, these agents are associated with numerous toxicities and long-term neurocognitive deficits in survivors. Our laboratory is interested in identifying drug resistance cell markers and combination therapies that target them to help increase efficacy of the chemotherapeutic agents. We have previously demonstrated that combination therapy of chemotherapeutic agents with Tolfenamic acid (TA), decreased the number of viable cancer cells when compared to the chemotherapeutic agents alone. TA is a non-steroidal anti-inflammatory drug, and its anti-cancer activity can be attributed to its ability to downregulate Specificity Protein 1 (Sp1), a transcription factor responsible for the upregulation of the anti-apoptotic protein, Survivin. Numerous cancerous tumors have been known to express high levels of Sp1 and SurvivIn, however these markers have not been well established in MB. Purpose: The aim of the project is to elucidate the association of Survivin expression in MB cells with the likelihood of patient survival and to test combination treatments of chemotherapeutic agents with the potential Survivin inhibitor, TA. The goal is to reduce the effective dosage of the chemotherapeutic agent Cisplatin, thereby reducing their side effect profiles. Methods: A R2 genomics visualization platform was accessed to obtain data regarding patient survival rates and Survivin expression in MB cells. A Kaplan-Meier curve was then generated to analyze the relationship. MB cell lines, DAOY, and D283 cells were obtained through the ATCC and cultured following standard cell culture conditions. Cells were then treated with vehicle (DMSO) or optimized doses of a chemotherapeutic agents (Vincristine or Cisplatin) with TA. Cell viability was measured at 24h and 48h post-treatment using Cell-TiterGlo kit (Promega) following manufacturer’s instructions. Results: The Kaplan-Meier curve showed that the overexpression of Survivin resulted in a poor prognosis and low survival rates among MB patients. Compared to results of MB cell inhibition with individual agents (Vincristine and Cisplatin) from our previous studies, the combination of TA and Vincristine or TA and Cisplatin showed decreased MB cell growth and downregulation of Survivin. Differential effects of Vincristine and Cisplatin were noted against DAOY and D283 cell lines. Conclusion: These preliminary observations suggest that Survivin expression may be associated with poor prognosis in MB patients and that inhibiting Survivin with TA is inducing the anti-proliferative effects of chemotherapeutic agents in MB cells. Further research involving other chemotherapies is required to understand TA’s role in Survivin inhibition. Understanding the specific effect of Cisplatin can help to design the therapies based on the molecular sub-group of MB.Item Specificity Protein 1 Expression and Cancer Patients Survival: Impact on Breast Cancer Patients Focusing on Race, Cancer Subclass, and Menopause(2024-03-21) Stock, Lauren; Dudhia, Ashni; Basha, RiyazPurpose: Transcription factors Specificity protein (Sp) 1 and 3 are known to regulate Baculoviral IAP Repeat-Containing 5 (BIRC5), an inhibitor of apoptosis protein, which is involved with poor prognosis and drug resistance in cancer therapy. The objective of this study is to determine the association of Sp1 expression with the prognosis of breast cancer patients using data from The Cancer Genome Atlas (TCGA) and determine the patient survival in relation to race, cancer subclass and menopause. Methods: Using TCGA data, Sp1 levels were monitored in tumor (n=1032) versus normal (non-tumor) breast tissue (n=114). Patient survival data was used to generate Kaplan Meier plots. Sp1 expression in non-tumor and tumor tissues and the association of Sp1 levels with the survival of patients with an emphasis on race, cancer subclass, menopause status and other factors were evaluated. Results: The analysis of TCGA data sets identified several cancers showing poor prognosis associated with higher expression of Sp transcription factors and BIRC5. The top cancers showing such association include adrenocortical carcinoma, brain lower grade glioma, chromophobe renal cell carcinoma, clear cell renal cell carcinoma, papillary renal cell carcinoma, hepatocellular carcinoma, pancreatic adenocarcinoma, rectal adenocarcinoma, sarcoma, and uterine corpus endometrial carcinoma. Breast cancer data showed poor survival rates with high expression of Sp1 and HER 2+ cancer type (p=0.0034). The expression of Sp1 showed significant variability amongst the Caucasian population in BRCA patient survival (p=0.046). While Sp1 expression is associated with poor survival rates in pre-, peri- and postmenopausal patients (p=0.02), the outcomes are worst among postmenopausal patients. Conclusions: The analysis of TCGA data sets showed a poor prognosis of cancer patients with high levels of Sp1, Sp3 and BIRC5. Overall survival is decreased with higher expression of these markers in multiple cancers including breast cancer. Furthermore, our findings suggest potential variations in prognosis based on factors such as race, cancer subclass, and menopausal status amongst breast cancer patients. Further research is needed to confirm the specific role of Sp transcription factors and BIRC5 in cancer patients’ survival focusing on critical variables such as age, sex, race and tumor stage.Item Targeting Sp1 in Ewing Sarcoma: A multi-approach method for the utilization of Mithramycin(2024-03-21) Lambring, Christoffer; Sankpal, Umesh; Behera, Santosh; Basha, RiyazPurpose: Ewing Sarcoma (ES) is a bone and soft tissue cancer affecting young adults and children. ES mostly occurs in the bones or soft tissue of the arms, legs, and pelvis. Localized ES presents with 5-year survival rate of 70%, but metastatic 5-year survival rate is between 15% and 30%. Our laboratory is interested in combination treatments using less toxic agents to induce sensitization to chemotherapy in ES. The anti-cancer activity of an antineoplastic antibiotic, Mithramycin, against ES cells has been shown. Mithramycin inhibits Specificity protein 1 (Sp1) a marker associated with aggressive cancer cell growth and resistance to chemo/radiation therapies. However, its mechanistic effects on other oncogenic proteins have yet to be elucidated in ES. The purpose of this study is to evaluate the effectiveness of Mithramycin and various combinations with other chemotherapeutic agents, Etoposide and Vincristine, to inhibit ES cell growth and assess the effect on key cancer related proteins regulated by Sp1. Future studies will include expanding upon Mithramycin’s mechanism of action in Ewing Sarcoma utilizing proteomics and various computational methods. Methods: Cell lines were obtained from Children’s Oncology Group (COG). Anti-proliferative activity of Mithramycin and/or Vincristine and Etoposide against ES cell lines, TC205 and CHLA10, was evaluated using CellTiterGlo kit. Dose curves were plotted and IC50 values were determined by Sigma-Plot software. The expression of Sp1 was determined by Western blot analysis. The specific type of effect (additive/antagonistic/ synergistic) of the combination treatments were determined by analyzing the combination index obtained via Calcusyn software. Nude mice were injected with TC205 cells and treated over two weeks with either Mithramycin (1mg/kg per week) and/or Etoposide (5mg/kg per week) and tumor volume was compared. Protein models were obtained from UnitProtKB and PDB and molecular dynamics simulations were run in the Schrödinger platform. Results: Mithramycin, etoposide, and vincristine decreased ES cell line viability in TC205 and CHLA10 cells as monotherapies, but more effectively in combination. Tumor volume was greatly attenuated upon Mithramycin and/or etoposide introduction, but more significantly when used in combination. Decreases in viability upon chemotherapeutic and Mithramycin introduction were drastically increased when used in combination and this effect was mirrored in further decreases in Sp1 expression. Synergistic drug responses were shown in the combination of Mithramycin with both Vincristine and Etoposide (CI <1). Sp1, Sp3, and survivin protein models were established and binding scoring and identification of key residues in Mithramycin protein interactions were identified. Conclusions: Mithramycin may effectively sensitize ES cells and improve the response of chemotherapy while lowering necessary effective dosages. Studies to understand the mechanism of action of Mithramycin on Sp1, Sp3, survivin, and other proteins involved in Ewing Sarcoma are underway.