Cancer
Permanent URI for this collectionhttps://hdl.handle.net/20.500.12503/21704
Browse
Browsing Cancer by Author "Basha, Riyaz"
Now showing 1 - 3 of 3
- Results Per Page
- Sort Options
Item In Vitro Study to Identify a Novel Combination Treatment for Ewing Sarcoma(2016-03-23) Sankpal, Umesh; Bowman, Paul; Ray, Anish; Conjeevaram Nagarajan, Bhavani Saranya; Basha, Riyaz; Gotlib, DanielIntroduction: Ewing sarcoma (ES) is an aggressive tumor that predominantly occurs in young adolescent populations. Chemotherapy used to treat ES is associated with long-term morbidity. The objective of this study was to identify an effective combination treatment option for treating ES. A fusion protein EWS-FLI1 is associated with [greater than] 85% of ES incidences and mithramycin (Mit) is known to target this fusion protein and is currently in clinical trials. We investigated the combination of Mit to induce the efficacy of Doxorubicin (Dox), a widely used chemotherapeutic agent for treating ES patients. Methods: ES cell lines CHLA9, CHLA10, and TC71 were cultured in laboratory and treated with increasing concentration of Mit or Dox. Cell viability was assessed using CellTiter-Glo luminescent assay at 48 hour post-treatment. Optimized doses for Mit and Dox were used for combination treatment. Western blot analysis was used to evaluate the apoptotic markers, caspase 3, cleaved PARP and Bcl2. Real-Time PCR was used to evaluate the expression of EWS-FL1 downstream targets. RNA was extracted using Trizol reagent (Invitrogen) and subjected to cDNA synthesis using Superscript III reverse transcriptase. PCR was carried out with cDNA using TaqMan primer-probes specific for ID2, LDB2, NROB1 and NCOR1. Results: The results of current study illustrated that Mit or Dox treatment resulted in a dose and time dependent decrease in ES cell proliferation. Mit down-regulated the EWS-FLI1 downstream targets (ID2, LDB2, NROB1 and RCOR1) in TC71 cells. Combination of Mit+Dox enhancesd the anti-proliferative effect in ES cell lines which was accompanied by modulation of apoptotic markers. Conclusion: This investigation provides evidence for the use of Mithramycin for enhancing the efficacy of chemotherapeutic agents. In vivo assays are underway to confirm in vitro results. These pre-clinical studies will have clinical implications for treating Ewing Sarcoma patients. The proposed combination to induce chemotherapy response is highly helpful since it can be tested to reducing chemotherapy dose(s) and addressing the issues related to side-effects.Item Tolfenamic Acid and Curcumin Analogs Effectively Inhibits Pancreatic Cancer Cell Growth(2016-03-23) Basha, Riyaz; Sankpal, Umesh; Hurtado, MyrnaTolfenamic Acid and Curcumin Analogs Effectively Inhibits Pancreatic Cancer Cell Growth Background: Curcumin (Cur) is a natural phenol of the plant Curcuma longa that has been tested for anti-cancer activity in pre-clinical and clinical studies. The therapeutic efficacy of Cur is greatly impacted by its low bioavailability. Synthetic analogs of Cur are under testing to overcome this limitation. Cur analogs, EF31 and UBS109 have shown anti-cancer activity in colon and pancreatic cancers. Small molecule Tolfenamic acid (TA) is known to inhibit human cancer cells and tumor growth in mouse models for some human cancers. TA is known to target Specificity protein 1 (Sp1) transcription factor that mediates the expression of several genes associated with cancer including survivin, a key member of inhibitor of apoptosis proteins family. Purpose: Combination treatments have been tested to address the issues related to efficacy and drug resistance. In this project, the anti-cancer activity of EF31 and UBS109 was tested in combination with TA. Method: Human pancreatic cancer cells, L3.6pl and MIA PaCa-2 were grown in DMEM media with 5% fetal bovine serum. Cells were treated with DMSO (control) or EF31 or UBS109 or TA or EF31+TA or UBS109+TA and cell viability was measured at 48 hour post treatment using CellTiter-Glo (Promega) kit. The effect on apoptosis was assessed by determining the activity of caspase 3/7 (Caspase-Glo kit) and the expression of cleaved PARP (Western blot). The effect of individual and combination treatment(s) on the expression of Sp1 and survivin was evaluated by Western blot analysis. Results: TA in combination with EF31 or UBS109 resulted in an induction of cell growth inhibition which is accompanied by increased caspase 3/7 activity and upregulation of PARP cleavage. The combination treatment(s) also showed a modulation in the expression of Sp1 and survivin. Conclusions: TA combination with EF31 or UBS109 increases pancreatic cancer cell growth inhibition potentially by inducing the apoptotic pathways. When compared to TA, TA and Cur analog(s) is more effective in pancreatic cancer cells. These new combinations of anti-cancer molecules provide preliminary evidence for the therapeutic efficacy in treating pancreatic cancer.Item Tolfenamic Acid Sensitizes Ewing Sarcoma Family Tumor Cells to Chemotherapy(2016-03-23) Sankpal, Umesh; Mike-Mayer, Austin; Tabor-Simecka, Leslie; Bowman, W. Paul; Ray, Anish; Basha, Riyaz; Shelake, SagarEwing sarcoma family tumor (EFT) is the second most prevalent bone and soft tissue tumor observed in children and young adolescents. Patients with metastatic disease have poor outcome with 5-year overall survival rate less than 30%. Current chemotherapeutic options are causing limited progress in the management of EFT. Our aim was to identify novel targets and less toxic agents to improve the efficacy of standard care offered to EFT patients. Specificity protein 1 (Sp1) transcription factor is known to be upregulated in various cancers and is frequently linked to poor prognosis. In this study, Tolfenamic acid, an inhibitor of Sp1 was tested to sensitize EFT cells to commonly used chemotherapeutic agent Vincristine (Vin). The effect of TA or Vin or TA+Vin on cell viability was evaluated by CellTiter-Glo assay and caspase 3/7 activity was measured by Caspase3/7 Glo assay. Western blot analysis was performed to determine the expression of Sp1, survivin, and cleaved-PARP. Apoptotic cell population was measured by Annexin V staining. Results showed a dose and time dependent decrease in cell viability with both agents while the combination of TA+Vin caused a significantly higher response as compared to individual treatments. This inhibition of cell viability was accompanied by the inhibition of Sp1 and survivin expression and an increase in the apoptotic markers i.e. Annexin-V positive cells, caspase 3/7 activity and the levels of c-PARP. Our results suggest that TA+Vin combination treatment provides an effective therapeutic strategy for the treatment of EFT.