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Frontiers in Chemistry 2023Despite modern sciences and advancements in new drugs or chemicals, the new era now rushes natural remedies for various illnesses and diseases that lead to end organ...
Despite modern sciences and advancements in new drugs or chemicals, the new era now rushes natural remedies for various illnesses and diseases that lead to end organ damage. In this study, we investigated ethanolic extract's effect against doxorubicin-induced cardiotoxicity and renal toxicity. To determine phytochemicals, a phytochemical screening was conducted. Various assays were used to measure the antioxidant activity, including the DPPH (2,2-diphenylpicrylhydrazyl), SOD (superoxide dismutase), NO (nitric oxide), and others. The antiproliferative effect of was assessed by MTT assay; morphological analysis was performed using an inverted and phase contrast microscope, ultra morphological analysis of apoptosis with acridine orange (AO)/propidium iodide (PI) staining. It was seen that doxorubicin caused elevated serum markers and abnormal changes in histological patterns. The significant reduction in cardiac and renal marker levels seen in groups given either 400 or 600 mg/kg of crude extract demonstrates that has a protective effect against doxorubicin-induced cardiotoxicity due to the presence of active phytoconstituents having antioxidant potential. There is a dose-dependent decrease in cell viability when using . Apoptosis was observed in the treated cells. In conclusion, our research lends credence to the idea that could be used for cancer management and have cardioprotective and nephroprotective effects.
PubMed: 38164252
DOI: 10.3389/fchem.2023.1283618 -
Cureus Nov 2023Increased colorectal carcinoma (CRC) and osteosarcoma prevalence, low survival rate, poor prognosis, and the limitations of existing anticancer therapies like side...
INTRODUCTION
Increased colorectal carcinoma (CRC) and osteosarcoma prevalence, low survival rate, poor prognosis, and the limitations of existing anticancer therapies like side effects of drugs, non-specificity, short half-life, etc., pose a need for novel anticancer drugs. Farnesol, an organic sesquiterpene compound, found in the essential oils of various plants has been shown to possess antioxidant, anti-inflammatory, and anticancer properties. However, the anticancer effect of farnesol against CRC and osteosarcoma has not yet been adequately elucidated.
AIM
The aim of the study was to analyze the anticancer effects of farnesol against human osteosarcoma and CRC cell lines.
MATERIALS AND METHODS
Human osteosarcoma (Saos-2) and colorectal carcinoma (HCT-116) cell lines were procured and cultured at 37C and 5% CO. The cells were treated with 10, 20, 40, 60, 80, and 100 µM/ml and 20, 40, 60, 80, 100, and 120 µM/ml of farnesol for 24 hours, respectively. 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide assay was performed to assess the cytotoxicity of farnesol on Saos-2 and HCT-116 cells. Acridine orange/ethidium bromide staining was carried out to analyze apoptosis. 4',6-diamidino-2-phenylindole staining was done to observe the nuclear changes. Dichloro-dihydro-fluorescein diacetate staining was performed to assess the farnesol-induced reactive oxygen species (ROS)-mediated cell death.
RESULTS
Farnesol reduced the viability and proliferation of Saos-2 and HCT-116 cells in a dose-dependent manner. Farnesol was able to alter the cellular and nuclear morphology of Saos-2 and HCT-116 cells, promoting cell death. Farnesol-induced apoptosis in human osteosarcoma and colorectal carcinoma cell lines. Early apoptosis was observed in farnesol-treated HCT-116 cells. Additionally, ROS-mediated apoptotic cell death was reported in Saos-2 cells.
CONCLUSION
Farnesol has the potential to induce cytotoxicity against human osteosarcoma and CRC cell lines.
PubMed: 38149135
DOI: 10.7759/cureus.49372 -
Cureus Nov 2023Aim The aim was to evaluate the anticancer potential of ethanolicleaf extract on MG-63 osteosarcoma cell lines. Materials and methods The anti-cancer properties of...
Aim The aim was to evaluate the anticancer potential of ethanolicleaf extract on MG-63 osteosarcoma cell lines. Materials and methods The anti-cancer properties of ethanolic leaf extract were evaluated on osteosarcoma cell lines using 3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and the morphological changes in MG-63 cells were assessed after 24 hours using microscopic observation. Additionally, fluorescence microscopy was employed to evaluate the apoptotic changes after acridine orange/ethidium bromide (AO/EtBr) dual staining. Results The MTT assay revealed a dose-dependent cell death. The cell viability decreased with increase in concentrations of the extract, The cell viability was 89.98 ± 4.89 percentage at 25 μg/ml and 15.64 ± 3.64 percentage at 200 μg/ml concentrations. A concentartion of 116.95 μg/ml showed 50% inhibition (IC). The morphological and dual staining studies also showed the extract's effectiveness in inducing apoptosis. Conclusion The ethanolic leaf extract of could impart good antiproliferative activity in MG-63 cell lines. The extract could also induce apoptosis and hence, it may be considered as a potential anticancer agent for the development of drug formulation for the treatment of osteosarcoma.
PubMed: 38143601
DOI: 10.7759/cureus.49276 -
FEBS Open Bio Mar 2024Exposure to time-varying electromagnetic fields (EMF) has the capacity to influence biological systems. Our results demonstrate that exposure to time-varying EMF modeled...
Exposure to time-varying electromagnetic fields (EMF) has the capacity to influence biological systems. Our results demonstrate that exposure to time-varying EMF modeled after the physiological firing frequency of intercellular calcium waves can inhibit proliferation and induce apoptosis in malignant cells. Single exposure of B16-BL6 cells to a Ca EMF for 40 min reduced the number of viable cells by 50.3%. Cell imaging with acridine orange and ethidium bromide dye revealed substantial cellular apoptosis, preapoptotic cells, nuclear fragmentation, and large spacing between cells in the Ca EMF condition when compared to the control condition. The ability of Ca EMF to influence the proliferation and survival of malignant cells suggests that exposure to specific EMF may function as a potential anticancer therapy.
Topics: Animals; Humans; Calcium Signaling; Electromagnetic Fields; Apoptosis; Time Factors; Melanoma, Experimental
PubMed: 38143305
DOI: 10.1002/2211-5463.13760 -
Medicina (Kaunas, Lithuania) Dec 2023: Cervical cancer is one of the most common types of frequently found cancers in Thailand. One of the causative agents is the infection of the high-risk human...
: Cervical cancer is one of the most common types of frequently found cancers in Thailand. One of the causative agents is the infection of the high-risk human papillomavirus (HPV) type 16 and 18. Traditional medicines are rich sources of bioactive compounds which are a valuable source for the development of novel cancer therapies. In this study, the therapeutic effects of 3 traditional medicines, Kerra, KS, and Minoza, were studied on HeLa and CaSki cells. : The effects of Kerra, KS, and Minoza on cancer cells were evaluated through cytotoxicity and cell death assays. The infection assay using HPV-16 pseudovirus was also carried out. : All traditional medicines efficiently suppressed cell growths of HeLa and CaSki, with Kerra being the most potent anticancer agent followed by KS and Minoza. Kerra at 158 µg/mL and 261 µg/mL significantly increases the percentage inhibition of the HPV-16 pseudovirus infection in a pre-attachment step in a dose-dependent manner, while KS at 261 µg/mL efficiently inhibited viral infection in both pre-attachment and adsorption steps. However, Kerra, KS, and Minoza at subtoxic concentrations could not reduce the viral E6 mRNA expressions of HPV-16 and HPV-18. Cell death assay by acridine orange/ethidium bromide showed that Kerra increased population of dead cells in dose-dependent manner in both CaSki and HeLa. The percentage of secondary necrosis in Kerra-treated CaSki was higher than that of HeLa cells, while the percentage of late apoptotic cells in HeLa was higher than that of CaSki, indicating that HeLa was more susceptible to Kerra than CaSki. For KS and Minoza, these extracts at 250 µg/mL promoted autophagy over cell death. At 500 µg/mL, the percentage of dead cells in Kerra was higher than that of KS and Minoza. : Kerra is a potent traditional medicine for promoting cancer cell death. Kerra is possibly useful in the prevention and treatment of cervical cancer. Further investigation will be carried out to gain a better understanding of the biochemical mechanism and the pharmacological activity underlying this effect.
Topics: Female; Humans; HeLa Cells; Papillomavirus Infections; Oncogene Proteins, Viral; Uterine Cervical Neoplasms; Antineoplastic Agents; Cell Line, Tumor; Apoptosis
PubMed: 38138272
DOI: 10.3390/medicina59122169 -
Frontiers in Oncology 2023Early-life osteosarcoma is associated with severe morbidity and mortality, particularly affecting young children and adults. The present cancer treatment regimen is...
INTRODUCTION
Early-life osteosarcoma is associated with severe morbidity and mortality, particularly affecting young children and adults. The present cancer treatment regimen is exceedingly costly, and medications like ifosfamide, doxorubicin, and cisplatin have unneeded negative effects on the body. With the introduction of hyphenated technology to create medications based on plant molecules, the application of ayurvedic medicine as a new dimension (formulation, active ingredients, and nanoparticles) in the modern period is rapidly growing. The primary source of lead compounds for the development of medications for avariety of ailments is plants and their products. Traditionally, (cumin) has been used as medication to treat a variety of illnesses and conditions.
METHODS
The cumin seed was successfully extracted with solvents Hexane, Chloroform, Methanol, Ethanol and Acetone. Following the solvent extraction, the extract residue was assayed in MG63 cells for their anti-proliferative properties.
RESULTS
First, we used the [3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide] (MTT) assay to test the extracted residue's cytotoxicity. The results show that hexane extract Half-maximal inhibitory concentration (IC50 86 µG/mL) effciently inhibits cells by causing programmed cell death. Furthermore, using the Acridine orange/ethidium bromide (AO/EB) staining method, the lactate dehydrogenase assay, and the reactive oxygen species assay using the Dichloro-dihydro-fluorescein diacetate (DCHFDA) staining method, we have demonstrated that the hexane extract causes apoptosis in MG63 cells. Furthermore, flow cytometry research revealed that the hexane extract stops the cell cycle in the S phase. In addition, the hexane extract limits colony formation and the migration potential as shown by the scratch wound healing assay. Furthermore, the extract from cumin seeds exhibits remarkable bactericidal properties against infections that are resistant to drugs. Gas chromatography analysis was used to quantitatively determine the hexane and methanolic extract based on the experimental data. The primary chemical components of the extract are revealed by the study, and these help the malignant cells heal. The present study finds that there is scientific validity in using cumin seeds as a novel method of anticancer therapy after undergoing both intrinsic and extrinsic research.
PubMed: 38125945
DOI: 10.3389/fonc.2023.1322875 -
Cytotherapy Feb 2024Accurate assessment of cell viability is crucial in cellular product manufacturing, yet selecting the appropriate viability assay presents challenges due to various...
BACKGROUND AIMS
Accurate assessment of cell viability is crucial in cellular product manufacturing, yet selecting the appropriate viability assay presents challenges due to various factors. This study compares and evaluates different viability assays on fresh and cryopreserved cellular products, including peripheral blood stem cell (PBSC) and peripheral blood mononuclear cell (PBMC) apheresis products, purified PBMCs and cultured chimeric antigen receptor and T-cell receptor-engineered T-cell products.
METHODS
Viability assays, including manual Trypan Blue exclusion, flow cytometry-based assays using 7-aminoactinomycin D (7-AAD) or propidium iodide (PI) direct staining or cell surface marker staining in conjunction with 7-AAD, Cellometer (Nexcelom Bioscience LLC, Lawrence, MA, USA) Acridine Orange/PI staining and Vi-CELL BLU Cell Viability Analyzer (Beckman Coulter, Inc, Brea, CA, USA), were evaluated. A viability standard was established using live and dead cell mixtures to assess the accuracy of these assays. Furthermore, precision assessment was conducted to determine the reproducibility of the viability assays. Additionally, the viability of individual cell populations from cryopreserved PBSC and PBMC apheresis products was examined.
RESULTS
All methods provided accurate viability measurements and generated consistent and reproducible viability data. The assessed viability assays were demonstrated to be reliable alternatives when evaluating the viability of fresh cellular products. However, cryopreserved products exhibited variability among the tested assays. Additionally, analyzing the viability of each subset of the cryopreserved PBSC and PBMC apheresis products revealed that T cells and granulocytes were more susceptible to the freeze-thaw process, showing decreased viability.
CONCLUSIONS
The study demonstrates the importance of careful assay selection, validation and standardization, particularly for assessing the viability of cryopreserved products. Given the complexity of cellular products, choosing a fit-for-purpose viability assay is essential.
Topics: Leukocytes, Mononuclear; Reproducibility of Results; Cell Survival; Trypan Blue; Cryopreservation; Flow Cytometry
PubMed: 38085197
DOI: 10.1016/j.jcyt.2023.11.008 -
Molecules (Basel, Switzerland) Nov 2023The objective of this study was to examine the preparation process of DSPE-PEG-C60/NCTD micelles and assess the impact of fullerenol (C60)-modified micelles on the...
Norcantharidin-Encapsulated C60-Modified Nanomicelles: A Potential Approach to Mitigate Cytotoxicity in Renal Cells and Simultaneously Enhance Anti-Tumor Activity in Hepatocellular Carcinoma Cells.
OBJECTIVE
The objective of this study was to examine the preparation process of DSPE-PEG-C60/NCTD micelles and assess the impact of fullerenol (C60)-modified micelles on the nephrotoxicity and antitumor activity of NCTD.
METHOD
The micelles containing NCTD were prepared using the ultrasonic method and subsequently optimized and characterized. The cytotoxicity of micelles loaded with NCTD was assessed using the CCK-8 method on human hepatoma cell lines HepG2 and BEL-7402, as well as normal cell lines HK-2 and L02. Acridine orange/ethidium bromide (AO/EB) double staining and flow cytometry were employed to assess the impact of NCTD-loaded micelles on the apoptosis of the HK-2 cells and the HepG2 cells. Additionally, JC-1 fluorescence was utilized to quantify the alterations in mitochondrial membrane potential. The generation of reactive oxygen species (ROS) following micelle treatment was determined through 2',7'-dichlorofluorescein diacetate (DCFDA) staining.
RESULTS
The particle size distribution of the DSPE-PEG-C60/NCTD micelles was determined to be 91.57 nm (PDI = 0.231). The zeta potential of the micelles was found to be -13.8 mV. The encapsulation efficiency was measured to be 91.9%. The in vitro release behavior of the micelles followed the Higuchi equation. Cellular experiments demonstrated a notable decrease in the toxicity of the C60-modified micelles against the HK-2 cells, accompanied by an augmented inhibitory effect on cancer cells. Compared to the free NCTD group, the DSPE-PEG-C60 micelles exhibited a decreased apoptosis rate (12%) for the HK-2 cell line, lower than the apoptosis rate observed in the NCTD group (36%) at an NCTD concentration of 75 μM. The rate of apoptosis in the HepG2 cells exhibited a significant increase (49%), surpassing the apoptosis rate observed in the NCTD group (24%) at a concentration of 150 μM NCTD. The HK-2 cells exhibited a reduction in intracellular ROS and an increase in mitochondrial membrane potential (ΔψM) upon exposure to C60-modified micelles compared to the NCTD group.
CONCLUSIONS
The DSPE-PEG-C60/NCTD micelles, as prepared in this study, demonstrated the ability to decrease cytotoxicity and ROS levels in normal renal cells (HK-2) in vitro. Additionally, these micelles showed an enhanced antitumor activity against human hepatocellular carcinoma cells (HepG2, BEL-7402).
Topics: Humans; Carcinoma, Hepatocellular; Liver Neoplasms; Micelles; Reactive Oxygen Species; Bridged Bicyclo Compounds, Heterocyclic; Cell Line, Tumor; Apoptosis
PubMed: 38005331
DOI: 10.3390/molecules28227609 -
International Journal of Molecular... Nov 2023The protein disulfide isomerase (PDI) family is a group of thioredoxin endoplasmic reticulum (ER)-resident enzymes and molecular chaperones that play crucial roles in...
The protein disulfide isomerase (PDI) family is a group of thioredoxin endoplasmic reticulum (ER)-resident enzymes and molecular chaperones that play crucial roles in the correct folding of proteins. PDIs are upregulated in multiple cancer types and are considered a novel target for cancer therapy. In this study, we found that a potent pan-PDI inhibitor, E64FC26, significantly decreased the proliferation of pancreatic ductal adenocarcinoma (PDAC) cells. As expected, E64FC26 treatment increased ER stress and the unfolded protein response (UPR), as evidenced by upregulation of glucose-regulated protein, 78-kDa (GRP78), phosphorylated (p)-PKR-like ER kinase (PERK), and p-eukaryotic initiation factor 2α (eIF2α). Persistent ER stress was found to lead to apoptosis, ferroptosis, and autophagy, all of which are dependent on lysosomal functions. First, there was little cleaved caspase-3 in E64FC26-treated cells according to Western blotting, but a higher dose of E64FC26 was needed to induce caspase activity. Then, E64FC26-induced cell death could be reversed by adding the iron chelator, deferoxamine, and the reactive oxygen species scavengers, ferrostatin-1 and N-acetylcysteine. Furthermore, the autophagosome-specific marker, light chain 3B (LC3B)-II, increased, but the autolysosome marker, sequestosome 1 (SQSTM1)/p62, was not degraded in E64FC26-treated cells. Using the FUW mCherry-LC3 plasmid and acridine orange staining, we also discovered a lower number of acidic vesicles, such as autolysosomes and mature lysosomes, in E64FC26-treated cells. Finally, E64FC26 treatment increased the cathepsin L precursor (pre-CTSL) but decreased mature CTSL expression according to Western blotting, indicating a defective lysosome. These results suggested that the PDI inhibitor, E64FC26, might initially impede proper folding of proteins, and then induce ER stress and disrupt proteostasis, subsequently leading to lysosomal defects. Due to defective lysosomes, the extents of apoptosis and ferroptosis were limited, and fusion with autophagosomes was blocked in E64FC26-treated cells. Blockade of autolysosomal formation further led to the autophagic cell death of PDAC cells.
Topics: Humans; Protein Disulfide-Isomerases; Proteostasis; Endoplasmic Reticulum Stress; Apoptosis; Pancreatic Neoplasms; Autophagy; Carcinoma, Pancreatic Ductal
PubMed: 38003657
DOI: 10.3390/ijms242216467 -
Biomedicines Nov 2023Phenol-soluble modulins (PSMs) are pore-forming toxins (PFTs) produced by staphylococci. PSMs exert diverse cellular effects, including lytic, pro-apoptotic,...
BACKGROUND
Phenol-soluble modulins (PSMs) are pore-forming toxins (PFTs) produced by staphylococci. PSMs exert diverse cellular effects, including lytic, pro-apoptotic, pro-inflammatory and antimicrobial actions. Since the effects of PSMs on autophagy have not yet been reported, we evaluated the autophagic activity in HaCaT keratinocytes treated with recombinant PSMα3.
METHODS
The autophagic flux and levels of autophagic marker proteins were determined using Western blot analysis. Subcellular localization of LC3B and Beclin-1 was investigated using an indirect immunofluorescence assay. The ultrastructural features of control and PSMα3-treated cells were evaluated via transmission electron microscopy. Cytoplasmic acidification was measured via acridine orange staining. Phosphorylation levels of protein kinases, implicated in autophagy regulation, were studied using a phospho-kinase array and Western blot analysis.
RESULTS
PSMα3 facilitated the intracellular redistribution of LC3B, increased the average number of autophagosomes per cell, promoted the development of acidic vesicular organelles, elevated the levels of LC3B-II, stimulated autophagic flux and triggered a significant decrease in the net autophagic turnover rate. PSMα3 induced the accumulation of autophagosomes/autolysosomes, amphisomes and multilamellar bodies at the 0.5, 6 and 24 h time points, respectively. The phospho-Akt1/2/3 (T308 and S473), and phospho-mTOR (S2448) levels were decreased, whereas the phospho-Erk1/2 (T202/Y204 and T185/Y187) level was increased in PSMα3-treated cells.
CONCLUSIONS
In HaCaT keratinocytes, PSMα3 stimulates autophagy. The increased autophagic activity elicited by sub-lytic PSM concentrations might be an integral part of the cellular defense mechanisms protecting skin homeostasis.
PubMed: 38002017
DOI: 10.3390/biomedicines11113018