Cancer
Permanent URI for this collectionhttps://hdl.handle.net/20.500.12503/21648
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Browsing Cancer by Author "Basha, Riyaz"
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Item Anti-cancer activity of biogenic silver nanoparticles against Neuroblastoma cells(2018-03-14) Umesh, Sankpal; Ravikumar, Nulakachandanam; Bharathkumar, Bukkapatnam; Basha, Riyaz; Rajasekhar, MaramABSTRACT Purpose: Neuroblastoma (NB) is one of the solid tumors diagnosed in young children. Due to severe side effects associated with the current therapeutic options, it is important to identify less toxic therapies for treating NB patients. Nanoparticles (NPs) are widely used in various medical applications; however, the particle size and preparation methods play critical roles in their activity. Recently use of plant extracts as stabilizing or reducing agents is gaining significance due to higher stability and activity. Biogenic silver nanoparticles (BSNPs) have been tested for their activity in wound healing mechanism and preventing microbial diseases. The objective of this investigation was to prepare BSNPs using plant extracts and silver nanoparticles and evaluate their anti-cancer activity against neuroblastoma (NB) cell lines. BSNPs were prepared using two different plant extracts and characterized. SHSY5Y and LA155n cells were treated with increasing concentrations of BSNPs for 48 h and dose curves obtained. The effect of BSNPs on apoptosis and cell cycle arrest was evaluated to understand the underlying mechanisms. Method: BSNPs were synthesized using silver NPs and herbal reducing agents. These particles were characterized by Atomic Force Microscope and Transmission Electron Microscope. Fourier-transform infrared spectroscopy was used to identify the active herbal compounds along with silver nanoparticles. The cell viability of NB cells was measured using Cell Titer-Glo kit. Apoptotic cells distribution were determined by Flow cytometry using annexin V staining The expression of cleaved Poly (ADP-ribose) Polymerase (cPARP) was evaluated by western blot. Results:. The characterization of BSNPs revealed alkynes, amines and alkylhalides as reducing agents and particles were ranged 20-100 nm in size. BSNPs caused significantly more cell growth inhibition when compared to silver NPs which is accompanied by an increase in apoptotic markers, c-PARP expression and annexin-v staining. Conclusion: These preliminary data using different reducing agents suggest the potential anti- proliferative effect of BSNPs against NB cells.Item Combination of Mithramycin and Standard Chemotherapeutic Agents Induces Anti-proliferative activity in Ewing Sarcoma cell lines(2018-03-14) Hunter, Abigail; Lout, Holly; Dunlap, Elissa; Sankpal, Umesh; Bowman, W. Paul; Basha, Riyaz; Ray, Anish; Albeer, LinaBackground/Hypothesis: Ewing sarcoma (ES) is a small, round, blue cell tumor found primarily in bones of adolescents. The EWS-FLI1 transcription factor is associated with proliferation of cancer cells and is over-expressed in [greater than] 85% of Ewing sarcoma cases. Mithramycin (MIT) is an antibiotic with antineoplastic properties and has been shown to inhibit EWS-FLI1. A recent trial of MIT treatment in ES patients found that hepatotoxicity precluded the administration of MIT at a dose required to inhibit EWS-FLI1 ([greater than]50nmol/L). We hypothesize that the efficacy of adjunct treatment can be enhanced if MIT is used along with standard chemotherapeutic agents such as Vincristine (VIN) and Etoposide (ETO). Combination treatment will reduce the effective dose of both Mithramycin and the standard agent thereby decreasing the therapeutic dose range and side effects. Methods: ES cells, CHLA10 and TC205 were cultured in the presence of vehicle or MIT or VIN or ETO or in combinations (MIT+VIN or MIT+ETO). After 2 days, cell viability was measured using The CellTiter-Glo® Luminescent Cell Viability Assay kit. The apoptosis induced by each of the above-mentioned treatments on the ES cells was measured by Flow cytometry using Annexin V Apoptosis Detection Kit. The expression of cleaved-Poly (ADP-ribose) polymerase (c-PARP), a marker for apoptosis was determined by Western blot analysis. Results: While all treatments showed ES cell growth inhibition, the combination treatment of MIT+ETO was more effective (significant at p Conclusion: The combination MIT+ETO caused more cell growth inhibition when compared to individual treatments in the TC205 and CHLA10 cell lines. These results demonstrate that MIT in combination with standard chemotherapeutic agents potentially increases therapeutic efficacy in ES. However, these results are limited to in vitro studies and need to be tested in an animal model to determine reproducibility and assess the toxicity.Item Copper Tolfenamic acid induces anti-proliferative activity effective against Medulloblastoma cells(2018-03-14) Sankpal, Umesh; Bowman, W. Paul; Basha, Riyaz; Arechiga, BiancaPurpose: Medulloblastoma (MB) is the most common pediatric malignant brain tumor, comprising 20% of all childhood brain tumors. Between 250-500 children per year are diagnosed in the US alone. Standard therapies result in severe long-term morbidities. Therefore, there is an urgent need for inventing novel effective treatment strategies with lower side-effects. Our laboratory showed anti-cancer activity of Tolfenamic acid (TA) in pre-clinical model for MB. Recent studies showed higher pharmacological effect of TA when synthesized as a complex with copper (Copper-TA, Cu-TA). Our aim was to investigate the anti-cancer activity of Cu-TA against MB cell lines. We hypothesize that Cu-TA presents higher anti-cancer activity and is more effective than TA to induce cytotoxicity against MB cells. Methods: DAOY and D283 cells were obtained from ATCC and grown following standard cell culture conditions. Cells were treated with TA or Cu-TA and the cell viability was measured at 24 and 48 h post-treatment using a CellTiter-Glo kit. The induction of apoptosis was investigated by studying caspase activation using the Caspase 3/7-Glo kit. In addition, reactive oxygen species (ROS) involvement was measured by flow cytometry. Results: Both Cu-TA and TA treatment resulted in decreased cell viability. However, when compared to TA, Cu-TA was more effective at inducing anti-proliferative activity in MB cells. Cu-TA induces increased production of ROS. The anti-proliferative activity of Cu-TA was accompanied by an increase in Caspase 3/7 activity, suggesting the induction of apoptosis. Conclusions: Cu-TA was more effective than TA. Therefore, it has potential as an effective anti-cancer agent for inhibiting MB cell growth. Further studies are needed to better understand Cu-TA’s mechanism of action.Item Evaluation of Metformin as an anti-cancer agent in Medulloblastoma(2018-03-14) Basha, Riyaz; Bowman, W. Paul; Sankpal, Umesh; Payne, KristenEvaluation of Metformin as an anti-cancer agent in Medulloblastoma Purpose: Medulloblastoma (MB) is the most common malignant brain tumor in children under 16 years of age. Standard treatment, including surgery, chemotherapy, and radiation, is successful for most; however, survivors often suffer from long-term neurocognitive and growth potential related sequelae. Therefore, there is a need to understand the molecular processes regulating MB growth to find less toxic therapies. Survivin is a protein in the Inhibitor of Apoptosis Protein (IAP) family that inhibits caspase activity. Survivin is highly expressed in MB and associated with a poor prognosis. Specificity protein 1 (Sp1) is a transcription factor regulating survivin expression and is overexpressed in many cancers. Interestingly, the use of Metformin (MET), an anti-diabetic drug, correlated with decreased occurrence of several cancers. Previous studies have demonstrated its anti-cancer activity in breast cancer cells as well. The objective of this study is to test the effect of MET on MB cells in vitro. Hypothesis: We hypothesize that MET treatment decreases the growth of MB cells in a dose and time-dependent manner, possibly inhibiting the expression of survivin via downregulating Sp1. Methods: DAOY (MB cell line from American Type Culture Collection) cells were treated with increasing concentrations of MET (0, 1, 5, 10, and 20 mM). Cell viability was assessed at 24 and 48 hours post-treatment using the CellTiter-Glo cell viability assay. Survivin and Sp1 expression in MET treated cells was determined by Western blot analysis. Potential mechanism of cell proliferation inhibition was investigated by measuring the induction of reactive oxygen species (ROS) through Flowcytometry. Results: MET treatment resulted in decreased cell viability in a dose and time dependent manner. MET treatment also decreased Sp1 and survivin expression indicating that the effect of MET is mediated via Sp1 transcription factor. We also observed MET induced cellular ROS formation, which could be a potential anti-cancer mechanism. Conclusion: Our data demonstrates that MET can inhibit MB cell growth, possibly via targeting Sp1 to down-regulate survivin and inducing ROS. We conclude that MET has the potential to be used in the treatment of MB. Due to limitations of using Metformin alone as an anti-cancer agent, additional experiments are underway to determine its use in conjunction with MB specific chemotherapeutic agents.Item Evaluation of Stability and Anti-cancer activity of Copper(II) Tolfenamic Acid with an emphasis on Pancreatic(2018-03-14) Sankpal, Umesh; Patel, Rafid; Chhabra, Jaya; Brown, Deondra; Gurung, Raj; Holder, Alvin; Rajasekhar, Maram; Basha, Riyaz; Hurtado, MyrnaPurpose: Tolfenamic acid (TA) acts as an anti-cancer agent in several cancer models via down-regulating transcription factors Sp1 and Sp3, and an inhibitor of apoptotic protein, survivin. Copper (Cu) is an important element with multiple biological functions and has gained interest in medical applications. Recently, Cu-TA has been synthesized and tested for enhanced therapeutic activity. In this study, Cu-TA was investigated for its stability and anti-cancer activity using several cancer cell lines and mouse model for pancreatic cancer (PC). Method: Cu-TA was synthesized and characterized by UV visible spectroscopy and Fourier-transform infrared spectroscopy (FTIR). Anti-proliferative activity was evaluated against twelve cell lines representing six (breast, colon, glioblastoma, medulloblastoma, pancreatic and prostate) cancers using the CellTiter-Glo kit and compared with TA. Further studies were performed using PC cells. The expression of Sp1, Sp3 and survivin was determined by Western blot and qPCR. The stability of Cu-TA was determined using 8-12 month-old powder and six-month-old stock solution. Cardiomyocytes (H9C2) were used to test the cytotoxicity in non-malignant cells. Athymic mice were injected with PC cells and treated with vehicle (control) or 25 or 50 mg/kg of Cu-TA 3 times/week and the effect on tumor growth was monitored for 4 weeks Animals body weight changes were also observed to determine overt toxicity. Results: Cu-TA significantly more effective than TA against all tested cancer cells. The IC50 values of Cu-TA were 30 to 80% less when compared with TA. Comparison of the twelve-month-old powder and six-month-old stock solution using the Panc1 cells showed similar IC50values ( Conclusion: These in vitro and in vivo studies demonstrate that Cu-TA is more effective than TA and potentially useful as an effective anti-cancer agent.Item Screening the ability of BNS-22 against chemotherapy-induced cytotoxicity in Cardiomyocytes(2018-03-14) Basha, Riyaz; Sankpal, Umesh; Hurtado, Myrna; Eskildsen, DaneHypothesis: Cardiac toxicity is one of the leading contraindications to many chemotherapeutic agents including anthracyclines (e.g. Doxorubicin). It has been demonstrated that knocking out the beta isozyme of topoisomerase II in mice results in amelioration of the cardiotoxic effects of Doxorubicin. The purpose of this study is to evaluate whether or not the inhibiton of the Topoisomerase II beta isozyme by the drug BNS-22 in cardiomyocytes can alleviate the cardiotoxic effects of doxorubicin. Methods/Materials: Cardiomyocyte cells (H9C2) were used to evaluate the cytotoxicity of BNS-22. Additionally, these cardiomyocytes were used to determine the rate of cardiac cell death in cells treated with Doxorubicin and BNS-22 concurrently compared to cells treated with Doxorubicin alone. Cell viability was measured by luminescence assay using the CellTiter-Glo kit. Cell viability was measured 72 hours after the administration of Vehicle (control) or BNS-22 or doxorubicin or doxorubicin and BNS-22. Results: Cardiomyocytes (H9C2) were grown following standard cell culture conditions. Cells which were treated with both Doxorubicin and BNS-22 together and the cells treated with only BNS-22 suffered considerably less cell loss than the cells treated with Doxorubicin alone. Conclusions: These preliminary results suggest that BNS-22 helps to alleviate the cardiotoxic effects of Doxorubicin. This experiment provides some evidence for the use of Topoisomerase inhibitors in the treatment of doxorubicin induced cardiotoxicity. Further cell viability assays using this drug will be performed to substantiate current findings.