Standard Chemotherapeutic Drugs Research Articles

Ferrocene-functionalized anilines as potent anticancer and antidiabetic agents: Synthesis, spectroscopic elucidation, and DFT calculations

Ferrocene derivatives have attracted significant interest as potent biological agents and novel drug candidates. Hence, the present research work was focused on the synthesis and characterization of ferrocene-integrated anilines (FB1-FB4) through different spectroscopic techniques such as: FT-IR, multinuclear (1H and 13C) NMR, Raman, atomic absorption spectroscopy, elemental analysis, and single-crystal X-ray crystallography. The crystallographic analysis revealed a supramolecular structure mediated by secondary non-covalent interactions (π—H and π—π). These ferrocenyl derivatives demonstrated a quasi-reversible electrochemical behavior with one electron transfer from Fe(II) to Fe(III)., which was evident by the change in the peak potential at different scan rates. The theoretically calculated energies of the HOMO-LUMO orbitals and the Mulliken charge distribution for the optimized structures determined by the DFT/B3LYP method associated well with the electrochemically measured redox potentials. The computational measurements also led to a close agreement between the found and observed vibrational frequencies having a correlation coefficient of 0.999. The in silico docking analysis against α-amylase, urease, and tyrosine kinase revealed that the docking scores are in close agreement with the in vitro obtained IC50 values of inhibitors. The synthesized compounds exhibited notable free radical scavenging activity against DPPH with IC50 values between 15.9–31.5 µM. The anticancer activity data revealed modest cytotoxicity against the subjected cancer cell line (MCF-7) compared with standard chemotherapeutic drug (cisplatin). However, these ferrocenyl analogs had fewer toxic effects in normal cells (MCF-10A). The results of the antidiabetic assay established that FB1 (IC50 = 19.27 µg/mL) is potentially more active inhibitor of α-amylase, the theoretical docking studies stating that FB1 is more prone towards hydrophobic interactions with Leu165 and Trp 59, in the active pocket of alpha-amylase. Together, the current study reveals the anti-cancer, anti-diabetic and anti-oxidant potential of newly synthesized ferrocene-substituted anilines.

Co-Expression of CD34, CD90, OV-6 and Cell-Surface Vimentin Defines Cancer Stem Cells of Hepatoblastoma, Which Are Affected by Hsp90 Inhibitor 17-AAG.

Cancer stem cells (CSCs) are nowadays one of the major focuses in tumor research since this subpopulation was revealed to be a great obstacle for successful treatment. The identification of CSCs in pediatric solid tumors harbors major challenges because of the immature character of these tumors. Here, we present CD34, CD90, OV-6 and cell-surface vimentin (csVimentin) as reliable markers to identify CSCs in hepatoblastoma cell lines. We were able to identify CSC characteristics for the subset of CD34+CD90+OV-6+csVimentin+-co-expressing cells, such as pluripotency, self-renewal, increased expression of EMT markers and migration. Treatment with Cisplatin as the standard chemotherapeutic drug in hepatoblastoma therapy further revealed the chemo-resistance of this subset, which is a main characteristic of CSCs. When we treated the cells with the Hsp90 inhibitor 17-AAG, we observed a significant reduction in the CSC subset. With our study, we identified CSCs of hepatoblastoma using CD34, CD90, OV-6 and csVimentin. This set of markers could be helpful to estimate the success of novel therapeutic approaches, as resistant CSCs are responsible for tumor relapses.

Formulation, Characterization and Cytotoxicity Effects of Novel Thymoquinone-PLGA-PF68 Nanoparticles.

Thymoquinone has anti-cancer properties. However, its application for clinical use is limited due to its volatile characteristics. The current study aims to develop a polymeric nanoformulation with PLGA-PEG and Pluronics F68 as encapsulants to conserve thymoquinone’s (TQ) biological activity before reaching the target sites. Synthesis of nanoparticles was successfully completed by encapsulating TQ with polymeric poly (D, L-lactide-co-glycolide)-block-poly (ethylene glycol) and Pluronics F68 (TQ-PLGA-PF68) using an emulsion–solvent evaporation technique. The size and encapsulation efficiency of TQ-PLGA-PF68 nanoparticles were 76.92 ± 27.38 nm and 94%, respectively. TQ released from these encapsulants showed a biphasic released pattern. Cytotoxicity activity showed that tamoxifen-resistant (TamR) MCF-7 breast cancer cells required a higher concentration of TQ-PLGA-PF68 nanoparticles than the parental MCF-7 cells to achieve IC50 (p < 0.05). The other two resistant subtypes (TamR UACC732 inflammatory breast carcinoma and paclitaxel-resistant (PacR) MDA-MB 231 triple-negative breast cell line) required a lower concentration of TQ-PLGA-PF68 nanoparticles compared to their respective parental cell lines (p < 0.05). These findings suggest that TQ encapsulation with PLGA-PEG and Pluronics F68 is a promising anti-cancer agent in mitigating breast cancer resistance to chemotherapeutics. In future studies, the anti-cancer activity of TQ-PLGA-PF68 with the standard chemotherapeutic drugs used for breast cancer treatment is recommended.

Substrate Stiffness Modulates the Growth, Phenotype, and Chemoresistance of Ovarian Cancer Cells.

Mechanical factors in the tumor microenvironment play an important role in response to a variety of cellular activities in cancer cells. Here, we utilized polyacrylamide hydrogels with varying physical parameters simulating tumor and metastatic target tissues to investigate the effect of substrate stiffness on the growth, phenotype, and chemotherapeutic response of ovarian cancer cells (OCCs). We found that increasing the substrate stiffness promoted the proliferation of SKOV-3 cells, an OCC cell line. This proliferation coincided with the nuclear translocation of the oncogene Yes-associated protein. Additionally, we found that substrate softening promoted elements of epithelial-mesenchymal transition (EMT), including mesenchymal cell shape changes, increase in vimentin expression, and decrease in E-cadherin and β-catenin expression. Growing evidence demonstrates that apart from contributing to cancer initiation and progression, EMT can promote chemotherapy resistance in ovarian cancer cells. Furthermore, we evaluated tumor response to standard chemotherapeutic drugs (cisplatin and paclitaxel) and found antiproliferation effects to be directly proportional to the stiffness of the substrate. Nanomechanical studies based on atomic force microscopy (AFM) have revealed that chemosensitivity and chemoresistance are related to cellular mechanical properties. The results of cellular elastic modulus measurements determined by AFM demonstrated that Young’s modulus of SKOV-3 cells grown on soft substrates was less than that of cells grown on stiff substrates. Gene expression analysis of SKOV-3 cells showed that mRNA expression can be greatly affected by substrate stiffness. Finally, immunocytochemistry analyses revealed an increase in multidrug resistance proteins, namely, ATP binding cassette subfamily B member 1 and member 4 (ABCB1 and ABCB4), in the cells grown on the soft gel resulting in resistance to chemotherapeutic drugs. In conclusion, our study may help in identification of effective targets for cancer therapy and improve our understanding of the mechanisms of cancer progression and chemoresistance.

Extracellular Sphingosine-1-Phosphate Downstream of EGFR Increases Human Glioblastoma Cell Survival.

Sphingosine-1-phosphate (S1P) is a crucial mediator involved in the progression of different cancers, including glioblastoma multiforme (GBM), the most frequent and deadly human brain tumor, characterized by extensive invasiveness and rapid cell growth. Most of GBMs overexpress the epidermal growth factor receptor (EGFR), and we investigated the possible link between S1P and EGFR signaling pathways, focusing on its role in GBM survival, using the U87MG human cell line overexpressing EGFR (EGFR+). We previously demonstrated that EGFR+ cells have higher levels of extracellular S1P and increased sphingosine kinase-1 (SK1) activity than empty vector expressing cells. Notably, we demonstrated that EGFR+ cells are resistant to temozolomide (TMZ), the standard chemotherapeutic drug in GBM treatment, and the inhibition of SK1 or S1P receptors made EGFR+ cells sensitive to TMZ; moreover, exogenous S1P reverted this effect, thus involving extracellular S1P as a survival signal in TMZ resistance in GBM cells. In addition, both PI3K/AKT and MAPK inhibitors markedly reduced cell survival, suggesting that the enhanced resistance to TMZ of EGFR+ cells is dependent on the increased S1P secretion, downstream of the EGFR-ERK-SK1-S1P pathway. Altogether, our study provides evidence of a functional link between S1P and EGFR signaling pathways enhancing the survival properties of GBM cells.

RSK Inhibition Induces Metaphase Arrest and Apoptosis in Acute Myeloid Leukemia Cells

The 90 kDa Ribosomal S6 Kinase (RSK) drives cell proliferation and survival in several cancers, although its oncogenic mechanism has not been well characterized. To examine the role of RSK in acute myeloid leukemia (AML), we analyzed apoptosis, DNA-damage, and the cell cycle profile following treatment with BI-D1870, a potent inhibitor of RSK. BI-D1870 treatment (5 µM, 12 h) of HL-60 and KG-1 cells increased the G2/M population and induced apoptosis. Previously, RSK has been shown to activate CDC2 for G2/M progression by dephosphorylating the negative regulatory Tyr14 through activation of CDC25 or inhibition of MYT1. Phosphorylation of CDC2 at Tyr14 was increased transiently during G2 phase following treatment with BI-D1870, while phosphorylation of the positive regulatory Ser198 of CDC25C was inhibited in G2 and mitotic populations. However, BI-D1870 treatment did not decrease CDC2/Cyclin B kinase activity, suggesting that inhibition of CDC2 is not a critical regulatory mechanism in BI-D1870-induced mitotic arrest. The mitotic population was further characterized by evaluation of expression levels of Cyclin A and Cyclin B with flow cytometry. Approximately 70% of mitotic cells accumulated in Cyclin A-negative/Cyclin B-high metaphase at 12 hours after 5 µM BI-D1870 treatment. Metaphase arrest was confirmed using a thymidine/nocodazole block to achieve synchronization at metaphase. Our results show that the metaphase/anaphase transition is significantly stalled by RSK inhibition as assessed by degradation of Cyclin B and Securin following release from mitotic arrest (Metaphase cells in mitotic population (%), control vs. BI-D1870, 0 h post-release: 80.53 ± 0.09% vs. 88.03 ± 0.67%; 2 h post-release: 56.27 ± 1.42% vs. 83.37 ± 0.32%; 4 h post-release: 23.53 ± 0.52% vs. 62.90 ± 0.62%, mean ± SEM, n=3, p< 0.01). We also assessed the efficacy of combination treatment of BI-D1870 with standard AML chemotherapeutic drugs cytarabine and daunorubicin. The combination of BI-D1870 with cytarabine or daunorubicin in HL-60 cells showed an additive effect. Furthermore, BI-D1870 induced apoptosis and metaphase arrest in primary AML cells and significantly suppressed the CD34+ subpopulation containing leukemia stem cells. Mitotic progression is controlled by the sequential degradation of cell cycle regulators including Cyclin A, Cyclin B, and Securin by the anaphase-promoting complex/cyclosome (APC/C) E3 ubiquitin ligase. Since proper association of CDC20 with APC/C is critical to destroy Cyclin B and Securin for metaphase/anaphase progression, we examined the effect of BI-D1870 on the complex formation of CDC20 with APC/C. Co-IP experiments showed that treatment of BI-D1870 impeded the association of activator CDC20 with APC/C, but increased binding of inhibitor MAD2 to APC/C, preventing metaphase/anaphase progression. These data show that BI-D1870 inhibits metaphase/anaphase progression by inactivating APC/C, resulting in apoptosis of AML cells. We are currently studying the effects of RSK knockdown on AML cells. In summary, our study demonstrates that RSK plays an important role in maintaining AML cell survival and proliferation, and RSK inhibitors could be used therapeutically to treat AML by inducing apoptosis and cell cycle arrest through the inhibition of APC/C in AML cells. DisclosuresNo relevant conflicts of interest to declare.

Dynamic cellular biomechanics in responses to chemotherapeutic drug in hypoxia probed by atomic force spectroscopy.

The changes in cellular structure play an important role in cancer cell development, progression, and metastasis. By exploiting single-cell, force spectroscopy methods, we probed biophysical and biomechanical kinetics (stiffness, morphology, roughness, adhesion) of brain, breast, prostate, and pancreatic cancer cells with standard chemotherapeutic drugs in normoxia and hypoxia over 12–24 hours. After exposure to the drugs, we found that brain, breast, and pancreatic cancer cells became approximately 55–75% less stiff, while prostate cancer cells became more stiff, due to either drug-induced disruption or reinforcement of cytoskeletal structure. However, the rate of the stiffness change decreased up to 2-folds in hypoxia, suggesting a correlation between cellular stiffness and drug resistance of cancer cells in hypoxic tumor microenvironment. Also, we observed significant changes in the cell body height, surface roughness, and cytoadhesion of cancer cells after exposure to drugs, which followed the trend of stiffness. Our results show that a degree of chemotherapeutic drug effects on biomechanical and biophysical properties of cancer cells is distinguishable in normoxia and hypoxia, which are correlated with alteration of cytoskeletal structure and integrity during drug-induced apoptotic process.

Mitochondrial ATP fuels ABC transporter-mediated drug efflux in cancer chemoresistance

Chemotherapy remains the standard of care for most cancers worldwide, however development of chemoresistance due to the presence of the drug-effluxing ATP binding cassette (ABC) transporters remains a significant problem. The development of safe and effective means to overcome chemoresistance is critical for achieving durable remissions in many cancer patients. We have investigated the energetic demands of ABC transporters in the context of the metabolic adaptations of chemoresistant cancer cells. Here we show that ABC transporters use mitochondrial-derived ATP as a source of energy to efflux drugs out of cancer cells. We further demonstrate that the loss of methylation-controlled J protein (MCJ) (also named DnaJC15), an endogenous negative regulator of mitochondrial respiration, in chemoresistant cancer cells boosts their ability to produce ATP from mitochondria and fuel ABC transporters. We have developed MCJ mimetics that can attenuate mitochondrial respiration and safely overcome chemoresistance in vitro and in vivo. Administration of MCJ mimetics in combination with standard chemotherapeutic drugs could therefore become an alternative strategy for treatment of multiple cancers.

Clinically-Relevant ABC Transporter for Anti-Cancer Drug Resistance

Multiple drug resistance (MDR), referring to the resistance of cancer cells to a broad spectrum of structurally and mechanistically unrelated drugs across membranes, severely impairs the response to chemotherapy and leads to chemotherapy failure. Overexpression of ATP binding cassette (ABC) transporters is a major contributing factor resulting in MDR, which can recognize and mediate the efflux of diverse drugs from cancer cells, thereby decreasing intracellular drug concentration. Therefore, modulators of ABC transporter could be used in combination with standard chemotherapeutic anticancer drugs to augment the therapeutic efficacy. This review summarizes the recent advances of important cancer-related ABC transporters, focusing on their physiological functions, structures, and the development of new compounds as ABC transporter inhibitors.

Abstract PO-011: Targeting colorectal cancer stem/progenitor cells with a combination of ionizing radiation and thymoquinone

Abstract Introduction: Radiotherapy is a standard neo-adjuvant therapy for cancer treatment, the success of which is limited by the resistance of cancer stem cells (CSCs) to therapy in addition to the deleterious effects of ionizing radiation (IR) to surrounding normal tissues. Despite the anticancer efficacy of combining IR with standard chemotherapeutic drugs, such treatments have shown great toxicity to normal cells. Thus, recent therapeutic strategies have aimed to combine IR with natural compounds that could radiosensitize and target cancer cells while sparing healthy tissues. The plant-derived compound Thymoquinone (TQ) has limited toxicity and is known to exert a radio-potentiating role in breast cancer and head and neck squamous cell carcinomas. Yet, no studies have investigated the antitumor effect of TQ in combination with IR on CSCs. Here, we studied the effect of TQ alone or in combination with IR on CSCs derived from different human colorectal cancer cell lines. Methods: MTT and trypan blue exclusion assays were used to study the effect of TQ and IR on proliferation and viability of HCT116 p53+/+, HCT116 p53−/−, and HT-29 p53+/− human colorectal cancer cell lines. The effect of TQ and IR on the long-term survival of different CRC cell lines was investigated by colony formation assay. Three-dimensional (3D) spheroid models derived from radiation resistant cells were used to enrich for stem/progenitor cells and study effects on CSCs self-renewal capacity. Results: IR doses of 1 or 2 Gray (Gy) significantly reduced the proliferation of HCT116 p53+/+ cells at 48 hours post IR. The antiproliferative effect of IR (2 Gy) on HCT116 p53−/− and HCT116 p53+/+ cells was significantly enhanced at 48 hours when combined with TQ at doses as low as 20 µM and 40 µM, respectively. While treatment of HCT116 p53+/+, HCT116 p53−/−, and HT-29 cells with 2 Gy IR or 10 µM TQ alone showed no significant inhibition on cell viability at 48 hours, combining the same doses of TQ and IR showed an enhanced and significant effect on cell viability when compared to TQ alone. Interestingly, combination treatment induced a more pronounced reduction in colony formation ability than either treatment alone at doses as low as 10 µM TQ. Of relevance to 3D systems, the combination of 3 µM TQ and 2 Gy IR significantly inhibited the self-renewal capacity of radiation resistant colonospheres derived from HT-29 cells at generation 1 (G1). Conclusion: Our study shows that the combination of IR with TQ appears to be a promising therapeutic strategy for eradicating radioresistant colorectal cancer cells and stem/progenitor cells. Citation Format: Samar Al Bitar, Farah Ballout, Wassim Abou-Kheir, Hala Gali-Muhtasib. Targeting colorectal cancer stem/progenitor cells with a combination of ionizing radiation and thymoquinone [abstract]. In: Proceedings of the AACR Virtual Special Conference on Radiation Science and Medicine; 2021 Mar 2-3. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(8_Suppl):Abstract nr PO-011.

Effects of Chemotherapy for Metastatic Colorectal Cancer on the TGF-β Signaling and Related miRNAs hsa-miR-17-5p, hsa-miR-21-5p and hsa-miR-93-5p.

Metastatic colorectal cancer (mCRC) patients are treated with standard chemotherapeutic drugs in the form of FOLFOX and FOLFIRI regimens. There are no reliable markers that could predict response to chemotherapy for mCRC. TGF-β signaling which interacts with microRNA (miRNA) network has important roles in tumor progression and chemotherapy resistance, thus the interplay between TGF-β signaling and miRNAs could be crucial for treatment response. The aim of this study was to analyze the effect of chemotherapy for mCRC on TGF-β signaling and related miRNAs. Hsa-miR-17-5p, hsa-miR-21-5p and hsa-miR-93-5p were selected out of 316 miRNAs with multiple targets within the TGF-β signaling by in silico analysis. SW620 cells were treated with chemotherapeutic drugs for mCRC for 1, 3 and 6 days and expression of selected miRNAs, PAI-1, CDH1 and VIM was measured. Expression of TGF-β signaling-related hsa-miR-17-5p, hsa-miR-21-5p and hsa-miR-93-5p was time-dependently altered in SW620 cells treated with chemotherapeutics for mCRC. The expression of hsa-miR-93-5p remained downregulated after 6 days under combined treatments FOX and FIRI as well as the hsa-miR-17-5p expression under FIRI. Chemotherapy regimens for mCRC increased expression of a major TGF-β signaling target gene PAI-1, independently of the selected miRNAs expression. These treatments also increased the expression of epithelial-mesenchymal transition (EMT) markers CDH1 and VIM on day 3 resulting in decrease of mesenchymal-like characteristics. However, their expression returned close to basal level on day 6. In conclusion, after initial response to chemotherapeutic drugs SW620 cells start to return close to the basal mesenchymal state while the long-term downregulated expression pattern of hsa-miR-93-5p and hsa-miR-17-5p makes them candidates worth testing as biomarkers for monitoring combined chemotherapeutic treatments therapy response in mCRC patients.

Bioinformatics analysis identified CDC20 as a potential drug target for cholangiocarcinoma.

BackgroundCholangiocarcinoma (CCA) is a malignancy that originates from bile duct cells. The incidence and mortality of CCA are very high especially in Southeast Asian countries. Moreover, most CCA patients have a very poor outcome. Presently, there are still no effective treatment regimens for CCA. The resistance to several standard chemotherapy drugs occurs frequently; thus, searching for a novel effective treatment for CCA is urgently needed.MethodsIn this study, comprehensive bioinformatics analyses for identification of novel target genes for CCA therapy based on three microarray gene expression profiles (GSE26566, GSE32225 and GSE76297) from the Gene Expression Omnibus (GEO) database were performed. Based on differentially expressed genes (DEGs), gene ontology and pathway enrichment analyses were performed. Protein-protein interactions (PPI) and hub gene identifications were analyzed using STRING and Cytoscape software. Then, the expression of candidate genes from bioinformatics analysis was measured in CCA cell lines using real time PCR. Finally, the anti-tumor activity of specific inhibitor against candidate genes were investigated in CCA cell lines cultured under 2-dimensional and 3-dimensional cell culture models.ResultsThe three microarray datasets exhibited an intersection consisting of 226 DEGs (124 up-regulated and 102 down-regulated genes) in CCA. DEGs were significantly enriched in cell cycle, hemostasis and metabolism pathways according to Reactome pathway analysis. In addition, 20 potential hub genes in CCA were identified using the protein-protein interaction (PPI) network and sub-PPI network analysis. Subsequently, CDC20 was identified as a potential novel targeted drug for CCA based on a drug prioritizing program. In addition, the anti-tumor activity of a potential CDC20 inhibitor, namely dinaciclib, was investigated in CCA cell lines. Dinaciclib demonstrated huge anti-tumor activity better than gemcitabine, the standard chemotherapeutic drug for CCA.ConclusionUsing integrated bioinformatics analysis, CDC20 was identified as a novel candidate therapeutic target for CCA.

Functional reconstruction of human AML reveals stem cell origin and vulnerability of treatment-resistant MLL-rearranged leukemia.

Chemoresistance remains the major challenge for successful treatment of acute myeloid leukemia (AML). Although recent mouse studies suggest that treatment response of genetically and immunophenotypically indistinguishable AML can be influenced by their different cells of origin, corresponding evidence in human disease is still largely lacking. By combining prospective disease modeling using highly purified human hematopoietic stem or progenitor cells with retrospective deconvolution study of leukemia stem cells (LSCs) from primary patient samples, we identified human hematopoietic stem cells (HSCs) and common myeloid progenitors (CMPs) as two distinctive origins of human AML driven by Mixed Lineage Leukemia (MLL) gene fusions (MLL-AML). Despite LSCs from either MLL-rearranged HSCs or MLL-rearranged CMPs having a mature CD34-/lo/CD38+ immunophenotype in both a humanized mouse model and primary patient samples, the resulting AML cells exhibited contrasting responses to chemotherapy. HSC-derived MLL-AML was highly resistant to chemotherapy and expressed elevated amounts of the multispecific anion transporter ABCC3. Inhibition of ABCC3 by shRNA-mediated knockdown or with small-molecule inhibitor fidaxomicin, currently used for diarrhea associated with Clostridium difficile infection, effectively resensitized HSC-derived MLL-AML toward standard chemotherapeutic drugs. This study not only functionally established two distinctive origins of human LSCs for MLL-AML and their role in mediating chemoresistance but also identified a potential therapeutic avenue for stem cell-associated treatment resistance by repurposing a well-tolerated antidiarrhea drug already used in the clinic.

In-Silico Analysis of Chromatin Modifiers and Profiling of Histone Deacetylases (HDAC's) in Human Oral Cancer.

Histone modifications have been demonstrated to play a significant role in oral squamous cell carcinoma (OSCC) epigenetic regulation. An in-silico analysis of The Cancer Genome Atlas (TCGA) of various histone acetyl transferases (HATs) and histone deacetylases (HDACs) suggested that HATs do not differ between normal and tumor samples whereas HDAC2 and HDAC1 change maximally and marginally respectively between normal and tumor patients with no change being noted in HDAC6 expression. Hence, this investigation was carried out to validate the expression states of HDAC 1, 2 and 6 mRNAs in buccal mucosa and tongue SCC samples in an Indian cohort. Buccal mucosa and tongue squamous cell carcinoma tissues with intact histopathology were processed for RNA isolation followed by cDNA synthesis which was then subjected to q-PCR for HDACs. The average RNA yield of the tongue tissue sample was ∼2 µg/mg of tissue and the A260/280 ratios were between 2.03 and 2.06. The average RNA yield of buccal mucosa tissue sample was ∼1 µg/mg of tissue and the A260/280 ratio were between 2.00 and 2.08. We have demonstrated that HDAC2 was overexpressed in tongue and buccal mucosa samples. Over-expression of HDAC2 imply potential use of HDACi along with standard chemotherapeutic drug in oral cancer treatment.

Evaluation of anti-cancer activity of Tinospora cordifolia on experimentally induced spontaneous fibrosarcoma in Swiss albino mice

Tinospora cordifolia, an Indian medicinal plant, was used to evaluate anti-cancer activity against spontaneous fibrosarcoma in Swiss albino mice. The qualitative and quantitative phytochemical analysis was done in hydroethanolic extract using physical tests and GC-MS study, respectively. Hundred Swiss albino female mice of eight week age were divided into 5 groups containing 20 mice each. Group A acted as normal control without tumor. In four groups B-E, fibrosarcoma was induced by tissue grafting method. Group B was kept without any treatment and served as tumor control. Group C was treated with hydroethanolic extract of T. cordifolia stem (at the rate 400mg/kg) for 30 days. Group D was treated with standard chemotherapeutic drug Paclitaxel (at the rate 5 mg/kg) alternate days. Group E was treated with both hydroethanolic extract of T. cordifolia stem (at the rate 400mg/ kg) for 30 days and Paclitaxel drug (5 mg/kg)in alternate days. We assessed the mean tumor volume, tumor doubling time, sur-vivability and percent increase in lifespan. These parameters revealed the anti-tumor activity of T. cordifolia against fibrosarcoma. In treatment groups, the histological alterations in neoplastic tissues were alternate patches of multiplying and necrotic tissues, vacuolation in tissues, and apoptotic changes. In T. cordifolia and combined treatment changes were more marked better than paclitaxel treatment. Post observational period upto 90 days revealed a good survival rate in combined treatment followed by T. cordifolia and paclitaxel treatment. The present data strongly suggested that the Tinosporacordifolia extract has anti-tumor potential and the combination therapy can be a good strategy to combat fibrosarcoma.

In Silico Examination of Peptides Containing Selenium and Ebselen Backbone to Assess Their Tumoricidal Potential

Cancer has been one of the highest causes of morbidity and mortality in the world for decades. More than 100 types of cancer have been identified with a wide range of treatment protocols comprising of chemotherapy, radiation therapy, hormone therapy, etc. In an attempt to curb the serious deleterious effects caused by the chemotherapeutic drugs, numerous peptide molecules are currently popular as alternatives to the standard chemotherapeutic drugs. In this study, we have carried out in silico investigations to ascertain the anti-proliferative potential of novel peptides based on selenium and ebselen, i.e. Eb-Trp-Asp, 13, Eb-Trp-Glu, 14, and Eb-Trp-Lys, 15. Analysis of protein-ligand interactions, resulting in protein-ligand complex formation, has been carried out using the AutoDockVina in PyRx aided molecular docking technique, which may be an essential indication of druggability of the test peptides. Findings suggested that novel peptide molecule Eb-Trp-Glu, 14 may be a potent anticancer agent.

Targeting the TGFβ pathway in uterine carcinosarcoma.

Uterine carcinosarcoma (UCS) is a relatively infrequent, but extremely aggressive endometrial malignancy. Although surgery and chemotherapy have improved outcomes, overall survival (OS) remains dismal due to the lack of targeted therapy and biphasic (epithelial and mesenchymal) nature that renders the tumor aggressive and difficult to manage. Here we report a role of transforming growth factor-β (TGFβ) in maintaining epithelial to mesenchymal transition (EMT) phenotype and aggressiveness in UCS. Using a 3D-culture system, we evaluated the efficacy of the transforming growth factor-β receptor-I (TGFβR1) kinase inhibitor Galunisertib (GLT), alone and in combination with standard chemotherapeutic drugs used for the management of UCS. We demonstrate that GLT by inhibiting canonical and non-canonical signaling emanating from transforming growth factor-β1 (TGFβ1) reduces cellular viability, invasion, clonal growth and differentiation. Interestingly, GLT sensitizes UCS cells to chemotherapy both in vitro and in in vivo preclinical tumor model. Hence, targeting TGFβ signaling, in combination with standard chemotherapy, may be exploited as an important strategy to manage the clinically challenging UCS.

Inhibition of Bruton's Tyrosine Kinase Suppresses Cancer Stemness and Promotes Carboplatin-induced Cytotoxicity Against Bladder Cancer Cells.

Bruton's tyrosine kinase (BTK) has been discovered to serve a critical role in the survival and infiltration of B-cell lymphoma. Recently, it was reported that BTK inhibitors exerted potential beneficial effects against numerous types of solid tumor, including glioblastoma multiforme and breast cancer; however, whether BTK is crucial for the progression of bladder cancer (BLCA) remains unclear. The present study investigated the in vitro function of BTK in stemness properties of BLCA cells. Furthermore, the therapeutic effects of a standard chemotherapeutic drug, carboplatin in combination with the BTK inhibitor, ibrutinib were also investigated. The association between BTK and BLCA progression was evaluated using free databases. The in vitro stemness and metastatic properties of BLCA cells were also investigated. Finally, the cytotoxicity of carboplatin in combination with ibrutinib was determined. The meta-survival analysis of the association between BTK and BLCA progression revealed that the expression levels of BTK were associated with a higher risk of BLCA progression. The CD133+-side population of BLCA cells formed spheroids when cultured in serum-free conditioned medium. In addition, expression levels of BTK and activated mTOR signaling in side population cells was up-regulated compared with the parental BLCA cells. Furthermore, the transfection of short hairpin RNA targeting BTK into BLCA cells markedly reduced cell migratory ability. More importantly, in advanced BLCA cells, which were more resistant to carboplatin, it was discovered that the cell viability was significantly reduced in the presence of ibrutinib (p<0.05). The findings of the present study suggested that BTK may have a critical role in the progression of BLCA; however, the underlying mechanisms and potential therapeutic strategies involved require further investigations.

Π electron-stabilized polymeric micelles potentiate docetaxel therapy in advanced-stage gastrointestinal cancer

Gastrointestinal (GI) cancers are among the most lethal malignancies. The treatment of advanced-stage GI cancer involves standard chemotherapeutic drugs, such as docetaxel, as well as targeted therapeutics and immunomodulatory agents, all of which are only moderately effective. We here show that Π electron-stabilized polymeric micelles based on PEG-b-p(HPMAm-Bz) can be loaded highly efficiently with docetaxel (loading capacity up to 23 wt%) and potentiate chemotherapy responses in multiple advanced-stage GI cancer mouse models. Complete cures and full tumor regression were achieved upon intravenously administering micellar docetaxel in subcutaneous gastric cancer cell line-derived xenografts (CDX), as well as in CDX models with intraperitoneal and lung metastases. Nanoformulated docetaxel also outperformed conventional docetaxel in a patient-derived xenograft (PDX) model, doubling the extent of tumor growth inhibition. Furthermore, micellar docetaxel modulated the tumor immune microenvironment in CDX and PDX tumors, increasing the ratio between M1-and M2-like macrophages, and toxicologically, it was found to be very well-tolerated. These findings demonstrate that Π electron-stabilized polymeric micelles loaded with docetaxel hold significant potential for the treatment of advanced-stage GI cancers.

Immunogenic chemotherapy in two mouse colon cancer models.

Aside from the induction of cell death, some anticancer chemotherapeutic drugs can modulate antitumor immune responses. In this study, we examined the anticancer effects of 5‐fluorouracil (5‐FU) and oxaliplatin (L‐OHP), which are standard chemotherapeutic drugs for colon cancer, combined with cyclophosphamide (CP) in two mouse colon cancer models (CT26 and MC38 colon adenocarcinoma models). In the CT26 model, two injections of 5‐FU/L‐OHP and CP significantly suppressed the growth of subcutaneously established CT26 tumors compared with either 5‐FU/L‐OHP or CP, without a significant loss of body weight. The anticancer effect was weakened in nude mice. Cured mice acquired protective immunity against CT26, and CT26‐specific cytotoxic T cells (CTLs) were induced from their spleen cells. Analysis of tumor‐infiltrating immune cells revealed that 5‐FU/L‐OHP treatment with or without CP increased the proportion of CD8+ T cells at tumor sites. The 5‐FU/L‐OHP treatment decreased the proportion of granulocytic myeloid‐derived suppressor cells (MDSCs) and increased monocytic MDSCs in tumor sites, whereas the addition of CP treatment reversed these changes. In the MC38 model, although significant anticancer effects of the triple combination therapy were seen, additional treatment with anti‐PD‐1 antibody increased the number of cured mice. These mice exhibited protective immunity against MC38, and MC38‐specific CTLs were generated from their spleen cells. Together, these results indicate that the antitumor effects of the combination of 5‐FU/L‐OHP and CP mainly depend on host T cells; moreover, the therapeutic efficacy can be effectively boosted by immune checkpoint blockade.