IC50 Values Of Doxorubicin Research Articles

Endogenous Magnetic Lipid Droplet-Mediated Cascade-Targeted Sonodynamic Therapy as an Approach to Reversing Breast Cancer Multidrug Resistance.

Multidrug resistance (MDR) has emerged as a major barrier to effective breast cancer treatment, contributing to high rates of chemotherapy failure and disease recurrence. There is thus a pressing need to overcome MDR and to facilitate the efficient and precise treatment of breast cancer in a targeted manner. In this study, endogenous functional lipid droplets (IR780@LDs-Fe3O4/OA) were developed and used to effectively overcome the limited diffusion distance of reactive oxygen species owing to their amenability to cascade-targeted delivery, thereby facilitating precise and effective sonodynamic therapy (SDT) for MDR breast cancer. Initially, IR780@LDs-Fe3O4/OA was efficiently enriched within tumor sites in a static magnetic field, achieving the visualization of tumor treatment. Subsequently, the cascade-targeted SDT combined with the Fenton effect induced lysosome membrane permeabilization and relieved lysosomal sequestration, thus elevating drug concentration at the target site. This treatment approach also suppressed ATP production, thereby inhibiting P-glycoprotein-mediated chemotherapeutic drug efflux. This cascade-targeted SDT strategy significantly increased the sensitivity of MDR cells to doxorubicin, increasing the IC50 value of doxorubicin by approximately 10-fold. Moreover, the cascade-targeted SDT also altered the gene expression profiles of MDR cells and suppressed the expression of MDR-related genes. In light of these promising results, the combination of cascade-targeted SDT and conventional chemotherapy holds great clinical promise as an effective treatment modality with excellent biocompatibility that can improve MDR breast cancer patient outcomes.

OIP5-AS1 enhances the malignant characteristics and resistance to chemotherapy of pancreatic cancer cells by targeting miR-30d-5p/MARCH8

MARCH8, an E3 ubiquitin ligase, plays a crucial role in regulating various cellular processes such as protein degradation and signaling pathways and is implicated in the development and spread of pancreatic cancer. Analysis of pancreatic cancer tissues compared to adjacent normal tissues showed a decrease in miRNA-30d-5p levels and an increase in OIP5-AS1 and MARCH8 levels, as confirmed by qRT-PCR and Western blot analysis. The dual-luciferase reporter assay demonstrated a binding relationship between OIP5-AS1 and miRNA-30d-5p, as well as between miRNA-30d-5p and MARCH8 in PACN-1 cells, derived from a human pancreatic carcinoma specimen. Further investigations utilizing various assays revealed that OIP5-AS1 inhibited apoptosis and promoted cell proliferation, invasion, and migration in PACN-1 cells via the miRNA-30d-5p/MARCH8 axis in vitro. Tumor experiments in nude mice confirmed that OIP5-AS1 enhanced PACN-1 cell growth in vivo through the miRNA-30d-5p/MARCH8 axis. Additionally, OIP5-AS1 was found to activate downstream genes of the JAK-STAT pathway, namely IFNAR2, SOCS3, and JAK1, in PACN-1 cells. Furthermore, OIP5-AS1 increased the IC50 values for doxorubicin, gemcitabine, and cisplatin in PACN-1 cells, as determined by the Cell Counting Kit-8 assay. Overall, OIP5-AS1 was shown to promote aggressive traits and resistance to chemotherapy in PACN-1 cells through the miRNA-30d-5p/MARCH8 axis.

Discovery of new 13(17)-epoxymyrsinane diterpenes from aerial flowering parts of euphorbia spinidens Bornm. ex Prokh. with proapoptotic activities against human bladder cancer EJ138 cells

Phytochemical analysis of aerial flowering parts of Euphorbia spinidens Bornm. ex Prokh. from Euphorbiaceae family (local name: Farfion-e-dandaneh-khari) led to the isolation of five diterpenes based on myrsinane backbone. Using HRESI-MS, 1D, and 2D NMR, they were identified as two previously unreported: 33,7,14,15(β)-tetraacetyl-5(α)-butanoyl-13α(17)epoxy-8,10(18)-myrsinadiene (1), 7,14,15(β)-triacetyl-3(β),5(α)-dibutanoyl-13α(17)epoxy-8,10(18)-myrsinadiene (2), and three known diterpenes: 3,7,14,15(β)-tetraacetyl-5(α)-propanoyl-13(17)-epoxy-8,10(18)-myrsinadiene (3), and 3,7,10,14,15(β)-Pentaacetyl-5(α)-butanoyl-13,17-epoxy-8-myrsinene (4), 3,7,10,14,15(β)-pentaacetyl-5(α)-propanoyl-13,17-epoxy-8-myrsinene (5). Compound 5 was previously reported in the roots of the same plant but without NMR data. Therefore, its mass pattern,1H-, and 13C-NMR data are reported. The cytotoxicity and proapoptotic properties of 1-3 were evaluated against EJ-138 bladder carcinoma cells through standard 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay for cytotoxicity screening and annexin V-FITC/PI apoptosis detection kit. In the cytotoxicity assay, the IC50 values found against EJ-138 were: (1) 41.6 ± 3.54 μM; (2) 38.4 ± 2.54 μM; (3) 57.3 ± 5.4 μM, whilst the IC50 value of doxorubicin was 1.7 ± 0.3 μM, respectively. In apoptosis assay, total apoptosis of compounds 1-3 at higher concentrations (100 μM) were 57.6 ± 3.54, 46.3 ± 2.82, and 57.2 ± 4.35%, respectively.

Enhancing Anticancer Potential: Investigating the Synergistic Impact of Doxorubicin and Curcumin on HeLa and Vero Cells in Vitro

(1) Background: Cervical cancer ranks as the fourth most prevalent cancer globally and in Asia, standing as the second most common in Indonesia. Despite its efficacy, doxorubicin chemotherapy is associated with significant side effects. To mitigate these adverse effects, a promising approach involves combining conventional drugs with curcumin. Both curcumin and doxorubicin have demonstrated cytotoxic effects against cervical cancer (HeLa). This research aims to determine the synergistic effect of the combination of doxorubicin and curcumin on HeLa and safety in normal cells (Vero cells). (2) Methods: This study adopted an experimental design utilizing doxorubicin and curcumin samples with HeLa and Vero cells. The investigation was initiated with cytotoxic and combination tests using the MTT method. The obtained results included IC50 values and combination indices, and the analysis involved a comparative examination of outcomes between HeLa and Vero cells. (3) Results: Cytotoxic tests revealed IC50 values for doxorubicin and curcumin on HeLa cells, measured in 2,17 ± 0,06 and 26,37 ± 2,00 µg/mL, and 17,32 ± 1,99 and 181,88 ± 30,48 µg/mL on Vero cells. Combination test results were represented by combination index .(4) Conclusions: The synergistic effect is observed at the combination of curcumin at a concentration of 9 μg/mL and doxorubicin at a concentration of 0,125 μg/mL, resulting in a combination index of 0,50. These findings suggest a promising avenue for enhancing the therapeutic potential of doxorubicin in cervical cancer treatment while minimizing adverse effects.

Combination therapy application of Abemaciclib with Doxorubicin in triple negative breast cancer cell line MDA-MB-231.

Due to lack of clinical biomarkers, Triple Negative Breast Cancer (TNBC) is more likely to have spread to other tissues at time of diagnosis and therapy planning generally involves use of cytotoxic chemotherapy agents, such as Doxorubicin. We aimed to investigate possible advantages of using combination strategy using Doxorubicin alongside Abemaciclib. After determining the IC50 values for Doxorubicin (DOX) and Abemaciclib (ABE); CompuSyn and ComBenefit software were used to reveal the effect resulting from the combination of two drugs. Following the determined effect, cell death was revealed by fluorescence microscopy and a colony forming assay was performed to see the potential of even a single cancer cell with adhesive character to survive over time and form a clone of itself. Detection of changing antioxidant activity following DOX, ABE and DOX+ABE combination therapy in MDAMB231 cells was determined by measuring MDA, SOD and GSH activities. The expression of Cleaved Caspase 3, PARP, Cleaved PARP, Cdk2 and Bax, which changed as a result of DOX, ABE and DOX+ABE application, was shown by Western Blotting.Cyclin-dependent kinase inhibitors appear as promising agents in therapy planning for breast cancer due to their prominent role in cell cycle regulation, where the number of studies interrogating its efficiency in the treatment of cancer such as TNBC is limited. For this reason, in this study, we aimed to determine the impact of the combined use of the CDK4/6 inhibitors ABE and DOX on the cytotoxicity, apoptotic homeostasis, alterations in antioxidative mechanisms, and the molecular pathways that they utilize. Our results showed that when used in combination, Doxorubicin and Abemaciclib showed a synergistic effect on TNBC cell line MDA-MB-231.

Effect of Trisindolina-5 Compound on Cancer Stem Cell (CSC) Proliferation in-Vitro

Cancer stem cells (CSCs) are a subset of cancer cells that have the abilities of normal stem cells. CSCs are cancer cell pioneers with self-renewal abilities that can cause CSCs to differentiate into several cancer cells. Because CSCs are resistant to conventional therapies, killing CSCs necessitates the use of a compound with powerful anticancer properties. Trisindoline has been shown to have powerful anticancer properties. Trisindoline has been synthesized into several modifications, the most recent of which is Trisindoline-5. The goal of this study is to find out what the IC50 value of Trisindoline-5 is. The cytotoxicity assay using Microculture Tetrazolium Technique Assay (MTT Assay) is used to determine IC50. The IC50 value of the Trisindoline-5 compound is 24.683 μM at 24 hours incubation, which classifies it as a medium cytotoxic compound, 17.067 μM at 48 hours incubation, which classifies it as a highly toxic compound, and 6497 μM at 72 hours incubation, which classifies it as a compound with no toxicity. While the IC50 value of doxorubicin is 1.611 μM after 24 hours, 2.334 μM after 48 hours, and 5.324 μM after 72 hours, it is classified as a compound with highly toxic activity.

In Vitro Evaluation of Cytotoxic and Antitumor Activities of The Tamoxifen and Doxorubicin Combination on MCF-7 and BT-474 Breast Cancer Cell Lines

The combination therapy of breast cancer has preferred for the patients to minimize possible side effects, drug resistance, recurrence and toxic effects. In this study, we aim to investigate the cytotoxic and antitumor activities the tamoxifen and doxorubicin combination in breast cancer cell lines, MCF-7 and BT-474. Tamoxifen (Tam) and doxorubicin (Dox) and their combination with different concentrations (0.625–20 μM Tam; 0.0625–2 μM Dox and 5 μM Tam+ 0.5/1.0/1.5 μM Dox combination were applied to MCF-7 and BT-474 cells for 48 hours. Afterthat, their cytotoxic activities were analyzed with MTT assay. Bcl-2, Mapt and Mrp1 are genes that induce cell proliferation, inhibit apoptosis and play role in drug resistance in cancer cells. To evaluate the antitumor activities of these genes in combination treatment, mRNA levels were analyzed by quantitative PCR. According to the MTT assay, it was determined that IC50 values as 17.26 μM and 16.65 μM for tamoxifen on MCF-7 and BT-474 breast cancer cell lines. IC50 values of doxorubicin in MCF-7 and BT-474 cells were 1.65 μM and 1.57 μM, respectively. It was found that the application of the combination drugs (15 μM tamoxifen and 1.5 μM doxorubicin) in MCF-7 and BT-474 cells have the lowest combination index values as 1.09 and 1.26, respectively. Therefore, the combination of 15 μM tamoxifen and 1.5 μM doxorubicin was selected and applied to both breast cancer cell lines for gene expression analysis. It was found that while Mrp1 and Mapt genes expressions were significantly upregulated, Bcl-2 gene expression was downregulated in MCF-7 cells. However, Bcl-2, Mrp1 and Mapt genes expressions in BT-474 cells were not significantly regulated. Altogether, these findings suggest that the combination of these two drugs may have a potential antagonistic interaction according combination index values.

A fluorescent system based on graphene quantum dots-capped magnetic hydroxyapatite-MIL-100 metal-organic frameworks for pH-sensitive and controlled release of DOX

Since metal-organic frameworks (MOFs) have shown great potential as emerging candidates in biomedical applications, it is worthwhile to construct an MOF-based drug delivery system. Hence, in this work, magnetic hydroxyapatite-MIL-100 metal-organic frameworks (SiO2@Fe3O4-HA-MIL-100) with antioxidant properties were synthesized as a novel drug delivery system through in-situ self-assembly of MIL-100 MOFs around pre-synthesized magnetic hydroxyapatite nanoparticles. Then, SiO2@Fe3O4-HA-MIL-100 nanocomposites were capped with fluorescent graphene quantum dots (SiO2@Fe3O4-HA-MIL-100-GQDs) for loading and delivering of doxorubicin (DOX). The SiO2@Fe3O4-HA-MIL-100, GQDs, and SiO2@Fe3O4-HA-MIL-100-GQDs encapsulation efficiency was 80.2, 42.2, and 95.8 %, respectively. The in vitro DOX release amount after 72 h from SiO2@Fe3O4-HA-MIL-100, GQDs, and SiO2@Fe3O4-HA-MIL-100-GQDs at pH 5 was 75.8 %, 86.2 %, and 67.2 %, while at pH 7.4 was 37.3 %, 50.1 %, and 29 %, respectively. These results confirmed a pH-responsive controlled release of DOX from nanocomposites. The release mechanism of DOX from the prepared nanocomposites was also well fitted to the Korsmeyer-Peppas kinetic, Fickian diffusion, and diffusion-controlled release. In addition, the MTT assays of nanocomposites against MCF-7 breast cancer cell lines clearly illustrated that the prepared nanocomposites had no significant cytotoxicity, while DOX-loaded nanocomposites had notable toxicity to MCF-7 cells. The IC50 values of DOX, SiO2@Fe3O4-HA-MIL-100-DOX, GQDs-DOX, and SiO2@Fe3O4-HA-MIL-100-GQDs-DOX against MCF-7 cells calculated at about 4.2 μg/mL, 1.8 μg/mL, 3 μg/mL, and 1.2 μg/mL, respectively. The DPPH test results also showed that the nanocomposites have excellent antioxidant activity. In addition, the in vitro hemolysis results displayed neglectable hemolysis activity (lower than 5 %) of SiO2@Fe3O4-HA-MIL-100-GQDs even at a high dose. Therefore, these new drug delivery systems can be used safely as potential drug carriers in targeted cancer therapy.

Synergistic combination of doxorubicin with hydralazine, and disulfiram against MCF-7 breast cancer cell line.

Disulfiram and hydralazine have recently been reported to have anti-cancer action, and repositioned to be used as adjuvant in cancer therapy. Chemotherapy combined with other medications, such as those that affect the immune system or epigenetic cell profile, can overcome resistance with fewer adverse effects compared to chemotherapy alone. In the present study, a combination of doxorubicin (DOX) with hydrazine (Hyd) and disulfiram (Dis), as a triple treatment, was evaluated against wild-type and DOX-resistant MCF-7 breast cancer cell line. Both wild-type MCF-7 cell line (MCF-7_WT) and DOX-resistant MCF-7 cell line (MCF-7_DoxR) were treated with different combination ratios of DOX, Dis, and Hyd followed by measuring the cell viability using the MTT assay. Synergism was determined using a combination index, isobologram analysis, and dose-reducing index. The anti-proliferation activity and mechanism of the triple combination were investigated by apoptosis analysis. The results showed a reduction in the IC50 values of DOX in MCF-7_WT cells (from 0.24 μM to 0.012 μM) and MCF-7_DoxR cells (from 1.13 μM to 0.44 μM) when treated with Dis (0.03μM), and Hyd (20μM) combination. Moreover, The triple combination DOX/Hyd/Dis induced significant apoptosis in both MCF-7_WT and MCF-7_DoxR cells compared to DOX alone. The triple combination of DOX, Dis, and Hyd showed a synergistic drugs combination to decrease the DOX dose needed to kill both MCF-7_WT and MCF-7_DoxR cancer cells and enhanced chemosensitivity to DOX.

Synthesis of Novel Chromene/[1,2,3]triazole Hybrid Derivatives: Cytotoxicity/Molecular Docking Studies

An easy and convenient approach has been adopted to synthesize a new series of chromene/[1,2,3]triazole hybrid derivatives involving Cu(I) catalyzed alkyne-azide 1,3-dipolar cycloaddition and a single step multicomponent reaction. The novel synthesized compounds (8a-l) were screened for cytotoxicity against three tumour cell lines i.e. MCF-7, PC-3 and HeLa using reference drug doxorubicin. Compound 8j (m-acetyl) showed an outstanding activity against all the three cell lines with IC50 values of 2.67 ± 0.03, 3.13 ± 0.03 and 3.05 ± 0.05 μM respectively. Compound 8k (p-acetyl) also exhibited good activity with IC50 values of 3.16 ± 0.05, 4.68 ± 0.03 and 3.81 ± 0.02 μM and the values are closer to doxorubicin IC50 values. The rest of the synthesized compounds have displayed good to moderate activity compared to reference drug. Molecular docking simulations of compounds 8b, 8f and 8j exhibited an excellent binding interactions against the crystal structure of epidermal growth factor receptor (EGFR).

Synergistic effect of the combination of Chrysanthemum cinerariifolium (Trev.) and doxorubicin in inhibiting PI3K and Cyclin D in oral squamous cell carcinoma: in vitro study

Background: The most common type of lips and oral cavity cancer is oral squamous cell carcinoma (OSCC). Doxorubicin (DX) is commonly used as a chemotherapy drug, but its use is limited due to risk factors and drug resistance. Chrysanthemum cinerariifolium (Trev.) (CC) has potential as an anticancer agent. Combining the plant extract and chemotherapy drug might prevent OSCC proliferation by inhibiting PI3K and cyclin D protein. Therefore, the present study aimed to determine the synergistic effect of the combination of C. cinerariifolium (Trev.) and doxorubicin in inhibiting PI3K and Cyclin D protein. Methods: Human oral squamous carcinoma cell lines SCC-9 were used in this study. A cytotoxicity assay was performed to obtain the IC50 value of CC ethanol extract and DX on the SCC-9 cell line. Synergism evaluation of the combination CC and DX was analyzed using CompuSyn software. ELISA and the immunofluorescent assay were performed to determine the level of PI3K and cyclin D in the SCC-9 cell line after being treated with IC50 value of CC, IC50 value of DX and three combinations of CC and DX [7/8 IC50 CC + 1/8 IC50 DX (dose 1), 6/8 IC50 CC + 2/8 IC50 DX (dose 2), and 4/8 IC50 CC + 4/8 IC50 DX (dose 3). Results: CC stem ethanol extract and DX inhibited the proliferation of SCC-9 cell lines with the IC50 value of 133.4 µg/mL and 288.3 nM, respectively. The combination of CC and DX at dose 2 (6/8 IC50 CC + 2/8 IC50 DX) exhibited a high decrease in PI3K and cyclin D expression. Conclusions: The combination of C. cinerariifolium and doxorubicin synergistically declined OSCC proliferation by inhibiting PI3K and cyclin D expression.

ZEB1 potentiates chemoresistance in breast cancer stem cells by evading apoptosis

Chemoresistance renders a challenge to the clinics to treat breast cancer patients. Current treatment strategies are effective in mitigating tumor growth but remain largely ineffective against cancer-initiating cells or breast Cancer Stem Cells (CSCs). Epithelial-to-mesenchymal-transition (EMT) regulates breast CSC physiology. Zinc finger E-box binding homeobox 1 (ZEB1) is a key EMT-transcription factor that regulates breast CSC - differentiation and metastasis. However, its potential role in modulating tumor chemoresistance has not yet been fully understood. In-silico analysis revealed a higher ZEB1 expression in breast cancer patients that leads to decreased overall and relapse-free survival. We generated sorted breast CSC with stable ZEB1 overexpression (CD24−/CD44+GFP-ZEB1) and/or silencing (CD24−/CD44+ZEB1 shRNA) as well as breast cancer cells with stable ZEB1 overexpression (CD24+GFP-ZEB1) and/or silencing (CD24+ZEB1 shRNA). An increased colony-forming efficiency and doxorubicin accumulation correlated with decreased promoter activity and expression profile of ABCC1 drug-efflux ABC transporter in CD24−/CD44+GFP-ZEB1. Additionally, CD24−/CD44+GFP-ZEB1 demonstrated doxorubicin-induced higher anti-apoptotic and lower pro-apoptotic protein expressions in the mitochondrial and cytosolic fractions. Chemoresistant CD24−/CD44+GFP-ZEB1 cells depicted 1000-fold higher IC-50 values of doxorubicin and decreased activation of JNK-p38 stress kinase molecular signaling-dependent mammosphere forming efficiency to evade apoptosis. Thus, ZEB1 and its downstream effectors are plausible therapeutic targets for the mitigation of breast cancer chemoresistance in patients.

Increased efficacy of biologics following inhibition of autophagy in A549 lung cancer cells in bimodal treatment of doxorubicin and SAR405-loaded chitosan nanoparticles

Conventional anti-cancer drugs, including doxorubicin, are associated with high toxicity and non-specific distribution in the body which results in a cascade of side effects. Cancer cells can utilize autophagy to promote tolerance to the stress induced by anti-cancer agents; this may be the major cause of drug resistance in advanced tumors. Owing to the molecular dynamism of cancer cells, targeting the pro-apoptotic survival pathways such as autophagy may increase the effectiveness of existing therapeutic agents at lower concentrations, thereby reducing the side effects of such agents. SAR405 is an inhibitor of autophagy activity due to its molecular interactions within the ATP binding site; however, it suffers from extracellular premature degradation and poor cellular uptake. Conversely, chitosan nanoparticles are considered biologically degradable, non-toxic, and biocompatible drug delivery agents that can be used to minimize the side effects of conventional therapeutic agents whilst increasing their intracellular uptake. In this study, a binary therapeutic strategy for the enhancement of the efficacy of doxorubicin while simultaneously inhibiting autophagy via a nano-mediated delivery system is reported. Autophagic inhibition was achieved via the improvement of SAR405 cellular uptake using SAR405-loaded chitosan nanoparticles. The synthesized nanoparticles were subsequently characterized for parameters of hydrodynamic diameter and polydispersity, while encapsulation and drug loading efficiencies were determined. Morphological characterization of the nanoparticles was elucidated using electron microscopy, and the cytotoxicity of the SAR405-loaded chitosan nanoparticles in combination treatments with doxorubicin was assessed through MTT and Annexin-v apoptosis assays. Autophagy progression through autophagosome formation was also evaluated using CYTO-ID staining. Following encapsulation, the size of the SAR405-loaded chitosan nanoparticles significantly increased from 54 nm to 161 nm at 10 µM SAR405 concentration, while the polydispersity index increased from 0.11 to 0.31 denoting presence of both encapsulated and unencapsulated moieties. When A549 lung cancer cells were treated with the IC50 values of doxorubicin in combination with SAR405-encapsulated CNP, an approximately 47% more reduction in cell viability was observed via the Annexin V-FITC/PI assay compared to using doxorubicin alone. Inhibition of autophagy was also detected in cells treated with SAR405 delivered using the nanoparticle system and was thought to be the primary reason towards a decrease in the resistance of the cancer cells to doxorubicin and thus increased its efficacy at lower concentrations. Therefore, this study has demonstrated a potential way of targeting cancer cell survival pathways that can be considered an effective way of increasing the efficacy of chemotherapeutic drugs.

Hyaluronan-Cyclodextrin Conjugates as Doxorubicin Delivery Systems.

In the last years, nanoparticles based on cyclodextrins have been widely investigated for the delivery of anticancer drugs. In this work, we synthesized nanoparticles with a hyaluronic acid backbone functionalized with cyclodextrins under green conditions. We functionalized hyaluronic acid with two different molecular weights (about 11 kDa and 45 kDa) to compare their behavior as doxorubicin delivery systems. We found that the new hyaluronan-cyclodextrin conjugates increased the water solubility of doxorubicin. Moreover, we tested the antiproliferative activity of doxorubicin in the presence of the new cyclodextrin polymers in SK-N-SH and SK-N-SH-PMA (over-expressing CD44 receptor) cancer cells. We found that hyaluronan-cyclodextrin conjugates improved the uptake and antiproliferative activity of doxorubicin in the SK-N-SH-PMA compared to the SK-N-SH cell line at the ratio 8/1 doxorubicin/polymer. Notably, the system based on hyaluronan (45 kDa) was more effective as a drug carrier and significantly reduced the IC50 value of doxorubicin by about 56%. We also found that hyaluronic acid polymers determined an improved antiproliferative activity of doxorubicin (IC50 values are on average reduced by about 70% of free DOXO) in both cell lines at the ratio 16/1 doxorubicin/polymer.

SYNTHESIS OF SILVER NANOPARTICLES USING ETHANOLIC EXTRACT OF Nelumbo nucifera Gaertn. LEAF AND ITS CYTOTOXIC ACTIVITY AGAINST T47D AND 4T1 CELL LINES

The cytotoxic activitiy of silver nanoparticles bio-functionalized by Nelumbo nucifera leaf extracts were examined in the current work. The nanoparticle (AgNPs) was characterized by scanning electron microscopy, UV–visible spectroscopy, and a particle size analyzer. Cytotoxic activity of silver nanoparticles against T47D and 4T1 cells using the MTT method using concentrations of 100 µg/mL, 50 µg/mL, and 25 µg/mL. For comparison, doxorubicin was used at the same dose. The size of the spherical nanoparticle was 66.4±0.93 nm and the polydispersity index was 0.23±0.05. Silver nanoparticles have anti-cancer properties against T47D and 4T1 cell lines based on inhibitory concentration. The inhibitory concentration (IC50) of AgNPs on T47D and 4T1 cells were obtained at 12.10±0.08 𝜇g/mL and 98.77±1.27 𝜇g/mL, respectively. Meanwhile, the IC50 values of doxorubicin in T47D and 4T1 cells were 4.45±0.03 𝜇g/mL and 36.77±1.15 𝜇g/mL, respectively. Overall, the results revealed that the green-synthetized silver nanoparticle had cytotoxic effects on breast cancer cells in comparison to doxorubicin.

Comparison of Characterization in Two-Dimensional and Three-Dimensional Canine Mammary Gland Tumor Cell Models.

Canine mammary gland tumors can be used as predictive models for human breast cancer. There are several types of microRNAs common in human breast cancer and canine mammary gland tumors. The functions of microRNAs in canine mammary gland tumors are not well understood. We compared the characterization of microRNA expression in two-dimensional and three-dimensional canine mammary gland tumor cell models. We evaluated the differences between two- and three-dimensional cultured canine mammary gland tumor SNP cells by assessing microRNA expression levels, morphology, drug sensitivity, and hypoxia. The expression of microRNA-210 in the three-dimensional-SNP cells was 10.19 times higher than that in the two-dimensional-SNP cells. The intracellular concentrations of doxorubicin in the two- and three-dimensional-SNP cells were 0.330 ± 0.013 and 0.290 ± 0.048 nM/mg protein, respectively. The IC50 values of doxorubicin for the two- and three-dimensional-SNP cells were 5.2 and 1.6 μM, respectively. Fluorescence of the hypoxia probe, LOX-1, was observed inside the sphere of three-dimensional-SNP cells without echinomycin but not in two-dimensional-SNP cells. The three-dimensional-SNP cells treated with echinomycin showed weak LOX-1 fluorescence. The present study showed a clear difference in microRNA expression levels in cells cultured in a two-dimensional adherent versus a three-dimensional spheroid model.

A simple, robust and scalable route to prepare sub-50 nm soft PDMS nanoparticles for intracellular delivery of anticancer drugs

Soft nanoparticles (NPs) have recently emerged as a promising material for intracellular drug delivery. In this regard, NPs derived from polydimethylsiloxane (PDMS), an FDA approved polymer can be a suitable alternative to conventional soft NPs due to their intrinsic organelle targeting ability. However, the available synthesis methods of PDMS NPs are complicated or require inorganic fillers, forming composite NPs and compromising their native softness. Herein, for the first time, we present a simple, robust and scalable strategy for preparation of virgin sub-50 nm PDMS NPs at room temperature. The NPs are soft in nature, hydrophobic and about 30 nm in size. They are stable in physiological medium for two months and biocompatible. The NPs have been successful in delivering anticancer drug doxorubicin to mitochondria and nucleus of cervical and breast cancer cells with more than four-fold decrease in IC50 value of doxorubicin as compared to its free form. Furthermore, evaluation of cytotoxicity in reactive oxygen species detection, DNA fragmentation, apoptosis-associated gene expression and tumor spheroid growth inhibition demonstrate the PDMS NPs to be an excellent candidate for delivery of anticancer drugs in mitochondria and nucleus of cancer cells.

Design, synthesis and molecular docking of new fused 1H-pyrroles, pyrrolo[3,2-d]pyrimidines and pyrrolo[3,2-e][1, 4]diazepine derivatives as potent EGFR/CDK2 inhibitors

A new series of 1H-pyrrole (6a–c, 8a–c), pyrrolo[3,2-d]pyrimidines (9a–c) and pyrrolo[3,2-e][1, 4]diazepines (11a–c) were designed and synthesised. These compounds were designed to have the essential pharmacophoric features of EGFR Inhibitors, they have shown anticancer activities against HCT116, MCF-7 and Hep3B cancer cells with IC50 values ranging from 0.009 to 2.195 µM. IC50 value of doxorubicin is 0.008 µM, compounds 9a and 9c showed IC50 values of 0.011 and 0.009 µM respectively against HCT-116 cells. Compound 8b exerted broad-spectrum activity against all tested cell lines with an IC50 value less than 0.05 µM. Compound 8b was evaluated against a panel of kinases. This compound potently inhibited CDK2/Cyclin A1, DYRK3 and GSK3 alpha kinases with 10–23% compared to imatinib (1–10%). It has also arrested the cell cycle of MCF-7 cells at the S phase. Its antiproliferative activity was further augmented by molecular docking into the active sites of EGFR and CDK2 cyclin A1.

Down-regulation of circITCH promotes osteosarcoma development and resistance to doxorubicin via the miR-524/RASSF6 axis.

Osteosarcoma (OS) is a malignant bone cancer, in which circular RNAs (circRNAs) act as important modulators. The present study aimed to explore the functional role of circRNA itchy E3 ubiquitin protein ligase (circITCH) in the development and doxorubicin (DXR) resistance of OS and the possible mechanistic pathway. A quantitative real-time polymerase chain reaction or western blot assays were exploited to analyze the expression of circITCH, miR-524 and Ras association domain family member 6 (RASSF6). Cell viability and half-maximal inhibitory concentration (IC50 ) value of DXR were monitored using a cell counting kit-8 assay. Cell migration, invasion and apoptosis were determined via a transwell assay and flow cytometry. The target interaction among circITCH, miR-524 and RASSF6 was validated by dual-luciferase reporter and RNA immunoprecipitation assays. A xenograft model of MG-63/DXR cells stably expressing circITCH in nude mice was established for assessing the role of circITCH in vivo. Down-regulation of circITCH and RASSF6, as well as the up-regulation of miR-524, was revealed in OS by investigating 40 paired OS tissue and normal tissue samples. Overexpression of circITCH lowered the cell viability, IC50 value of DXR, migration and invasion, whereas it facilitated apoptosis of OS cells. circITCH sponged miR-524 to up-regulate RASSF6, causing OS progression inhibition and DXR resistance reduction. Additionally, circITCH up-regulation reduced tumor growth in vivo. Transduction with circITCH represses OS progression and promotes DXR sensitivity by the miR-524/RASSF6 axis, providing a new perspective for therapeutic intervention.

De novo synthesis of pH-responsive, self-assembled, and targeted polypeptide nano-micelles for enhanced delivery of doxorubicin

Doxorubicin (DOX) is a commonly used anticancer drug, but it is inefficient as a therapeutic due to a lack of targeting. Peptide-tuned self-assembly of DOX offers a strategy to improve targeting for greater efficacy. In this work, we designed and prepared an amphiphilic tumor cell-targeting peptide, P14 (AAAAFFFHHHGRGD), able to encapsulate DOX by self-assembly to form tumor cell-targeting and pH-sensitive nano-micelles. The results showed a critical P14-micelle concentration of 1.758 mg l−1 and an average particle size of micelles of 121.64 nm, with entrapment and drug-loading efficiencies of 28.02% ± 1.35% and 12.06% ± 0.59%, respectively. The prepared micelles can release 73.52 ± 1.27% DOX within 24 h in pH 4.5 medium, and the drug cumulative release profile of micelles can be described by the first-order model. Compared with free DOX, the micelles exhibited an increased ability to inhibit tumor cell growth and cause tumor apoptosis in vitro, with IC50 values of DOX and P14-DOX micelles against human breast cancer cells (MCF-7) of 0.91 ± 0.07 and 0.75 ± 0.06 μg ml−1, respectively, and cellular apoptotic rates of DOX and P14-DOX micelles of 70.3% and 42.4%, respectively. Cellular uptake experiments revealed high concentrations of micelles around and inside MCF-7 cells, demonstrating that micelles can target tumor cells. These results indicate the excellent potential for the application of this amphiphilic peptide as a carrier for small-molecule drugs and suggest a strategy for the design of effective anti-tumor drugs.