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Journal of Cancer Research and... Oct 2023GW9508, a free fatty acid receptor agonist acts in a G-coupled protein receptor 40 (GPR40)-dependent pathway. Here, we investigated the induction of stress oxidative and...
AIMS
GW9508, a free fatty acid receptor agonist acts in a G-coupled protein receptor 40 (GPR40)-dependent pathway. Here, we investigated the induction of stress oxidative and autophagy by GW9508 in the human colorectal cancer cell line (HT-29) and the crosstalk between autophagy and apoptotic in HT-29 cells.
METHODS
HT-29 was treated with GW9508 at a concentrations range of 50-500 μM in fibrin gel. Cell viability was investigated using an MTT assay. Induction of autophagy and apoptosis was assessed through Western blotting for associated proteins, acridine orange staining, MDC staining, qRT-PCR, and electron microscopy. Also, we estimated the molecular interactions between GW9805 and some markers through molecular docking.
RESULTS
GW9508 inhibited HT-29 cell proliferation, induced apoptosis, and resulted in autophagy. The induced autophagy in cells was confirmed by the observation of autophagosomes, the presence of autophagy markers, including beclin-1, LC3, AMPK, and lack expression of mTOR and AKT. Moreover, GW9508 treatment significantly increased the expression of catalase and superoxide dismutase in cells.
DISCUSSION
Our results indicated that GW9508 could induce autophagy by inhibiting the Akt/mTOR in HT-29. Hence, GW9508 is suggested as a novel anticancer reagent.
Topics: Humans; Autophagy; HT29 Cells; Methylamines; Molecular Docking Simulation; Propionates; Proto-Oncogene Proteins c-akt; Signal Transduction; TOR Serine-Threonine Kinases; Receptors, Cell Surface
PubMed: 38376299
DOI: 10.4103/jcrt.jcrt_1184_21 -
Cancer Biology & Therapy Dec 2024The antipsychotic drug pimozide has been demonstrated to inhibit cancer. However, the precise anti-cancer mechanism of pimozide remains unclear. The purpose of this...
The antipsychotic drug pimozide has been demonstrated to inhibit cancer. However, the precise anti-cancer mechanism of pimozide remains unclear. The purpose of this study was to investigate the effects of pimozide on human MCF-7 and MDA-MB-231 breast cancer cell lines, and the potential involvement in the RAF/ERK signaling. The effects of pimozide on cells were examined by 4,5-dimethylthiazol-2-yl-3,5-diphenylformazan, wound healing, colony formation, transwell assays, and caspase activity assay. Flow cytometry and acridine orange and ethidium bromide staining were performed to assess changes in cells. Transmission electron microscopy and monodansylcadaverine staining were used to observe autophagosomes. The cyclic adenosine monophosphate was evaluated using the FRET system. Immunohistochemistry, immunofluorescence, RNA interference, and western blot investigated the expression of proteins. Mechanistically, we focus on the RAF1/ERK signaling. We detected pimozide was docked to RAF1 by Schrodinger software. Pimozide down-regulated the phosphorylation of RAF1, ERK 1/2, Bcl-2, and Bcl-xl, up-regulated Bax, and cleaved caspase-9 to induce apoptosis. Pimozide might promote autophagy by up-regulating cAMP. The enhancement of autophagy increased the conversion of LC3-I to LC3-II and down-regulated p62 expression. But mTOR signaling was not involved in promoting autophagy. The knockdown of RAF1 expression induced autophagy and apoptosis in breast cancer cells, consistent with the results of pimozide or sorafenib alone. Blocked autophagy by chloroquine resulted in the impairment of pimozide-induced apoptosis. These data showed that pimozide inhibits breast cancer by regulating the RAF/ERK signaling pathway and might activate cAMP-induced autophagy to promote apoptosis and it may be a potential drug for breast cancer treatment.
Topics: Humans; Female; MAP Kinase Signaling System; Breast Neoplasms; Antipsychotic Agents; Pimozide; Cell Proliferation; Apoptosis; Autophagy; Cell Line, Tumor
PubMed: 38356266
DOI: 10.1080/15384047.2024.2302413 -
Saudi Pharmaceutical Journal : SPJ :... Mar 2024is traditionally used to treat breast cancer in several Arab countries. Scientific studies have reported different effects of this plant on some cancer cell lines. The...
is traditionally used to treat breast cancer in several Arab countries. Scientific studies have reported different effects of this plant on some cancer cell lines. The current study determined the anti-cancer potential of the methanolic extract of against four different types of breast cancer cell lines . The extract was prepared by maceration and phytoconstituents were identified by LC-MS analysis. The IC value was determined against MDA-MB-231, MCF-7, 4 T1, and MCF-10 cell lines using the MTT assay. Further investigations were carried out using IC concentration of the extract (40.09 µg/ml) to determine live/dead cells by acridine orange/ethidium bromide staining. The effect on the expression of reactive oxygen species (ROS) was evaluated by flow cytometry. The results were analyzed using one-way ANOVA followed by Tukey's test. The LC-MS analysis revealed the presence of 34 and 30 phytoconstituents in positive and negative modes respectively. The extract was most effective against 4 T1 cells in a dose-dependent manner (P < 0.001) with an IC value of 40.09 µg/ml and showed negligible effect against MCF-10 cells. It increased apoptosis in 77.84 % of 4 T1 cells, as determined by acridine orange/ethidium bromide staining. The extract also increased the ROS expression in the 39.57 % of 4 T1 cells. The study results showed that Ephedra foeminea extract possesses an anti-cancer effect against 4 T1 cells by increasing the expression of ROS and inducing apoptosis in the 4 T1 cells. The result suggests methanolic extract possesses a reasonable anti-cancer effect due to its effect on apoptosis and oxidative pathways. The results confirm the traditional belief that is effective against breast cancerز.
PubMed: 38328794
DOI: 10.1016/j.jsps.2024.101960 -
International Journal of Biological... 2024As lung cancer is the leading cause of cancer death worldwide, the development of new medicines is a crucial endeavor. Naringenin, a flavanone derivative, possesses...
As lung cancer is the leading cause of cancer death worldwide, the development of new medicines is a crucial endeavor. Naringenin, a flavanone derivative, possesses anti-cancer and anti-inflammatory properties and has been reported to have cytotoxic effects on various cancer cells. The current study investigated the underlying molecular mechanism by which naringenin induces cell death in lung cancer. The expression of apoptosis, cell cycle arrest, and autophagy markers in H1299 and A459 lung cancer cells was evaluated using a terminal deoxynucleotidyl transferase dUTP nick end labeling assay (TUNEL), Western blot, Annexin V/PI stain, PI stain, acridine orange staining, and transmission electron microscopy (TEM). Using fluorescence microscopy, DALGreen was used to observe the degradation of p62, a GFP-LC3 plasmid was used to evaluate puncta formation, and a pcDNA3-GFP-LC3-RFP-LC3ΔG plasmid was used to evaluate autophagy flux. Furthermore, the anti-cancer effect of naringenin was evaluated in a subcutaneous H1299 cell xenograft model. Naringenin treatment of lung cancer cells (H1299 and A459) reduced cell viability and induced cell cycle arrest. Pretreatment of cells with ROS scavengers (-acetylcysteine or catalase) suppressed the naringenin-induced cleavage of apoptotic protein and restored cyclin-dependent kinase activity. Naringenin also triggered autophagy by mediating ROS generation, thereby activating AMP-activated protein kinase (AMPK) signaling. ROS inhibition not only inhibited naringenin-induced autophagic puncta formation but also decreased the ratio of microtubule-associated proteins 1A/1B light chain 3 II (LC3II)/LC3I and activity of the AMPK signaling pathway. Furthermore, naringenin suppressed tumor growth and promoted apoptosis in the xenograft mouse model. This study demonstrated the potent anti-cancer effects of naringenin on lung cancer cells, thereby providing valuable insights for developing small-molecule drugs that can induce cell cycle arrest, apoptosis, and autophagic cell death.
Topics: Humans; Animals; Mice; Carcinoma, Non-Small-Cell Lung; Apoptosis; Lung Neoplasms; Reactive Oxygen Species; AMP-Activated Protein Kinases; Cell Line, Tumor; G2 Phase Cell Cycle Checkpoints; Autophagy; Flavanones
PubMed: 38322119
DOI: 10.7150/ijbs.85443 -
Journal of Oral and Maxillofacial... 2023Sex determination in forensic medicine is considered one of the first and foremost steps in personal identification. The need for identifying the exact sex of the...
UNLABELLED
Sex determination in forensic medicine is considered one of the first and foremost steps in personal identification. The need for identifying the exact sex of the individual arises when deciding whether a person can exercise certain civil rights reserved for one particular sex, for competing in sex-specific athletic and sports events, legitimacy, divorce, paternity disputes and also to some criminal offenses. Nuclear sexing by Barr body examination can be done using buccal smears to establish the sex of the individual when routine methods fail to disclose the exact gender of the individual.
AIM
To determine and compare the Barr bodies present in exfoliated buccal epithelial cells in males, females and transgender populations using light and fluorescence microscopy.
MATERIALS AND METHODS
A total of 90 patients were recruited for the study. Group I consisted of 30 female patients. Group II consisted of 30 male patients and group III consisted of 30 transgender patients. The buccal mucosa was then scraped using a wooden spatula and the cells obtained were fixed in 95% ethanol. Two smears per individual were made and stained. One smear was stained with papanicolaou (PAP) stain and the other with Acridine orange and viewed under light microscopy and fluorescent microscopy, respectively.
RESULTS
When PAP stained slides were examined, the percentage of Barr-bodies in females ranged from 3% to 5% and in males it was 0% and in transgenders, it ranged from 0% to 5%. In Acridine orange stained smears, the percentage of Barr bodies in females ranged from 1% to 3% and in males it was 0% and in transgenders, it was 0%. Kruskal-Wallis test to study the relation of Barr body percentage in females, males and transgender subjects demonstrated significant differences between the groups ( < 0.001). Wilcoxon signed rank test was done for pairwise comparison, which showed that the distribution of percentage of positive cells in females are statistically significant from males and transgenders ( < 0. 001).
CONCLUSION
Nuclear sexing using Barr bodies offers a simple yet effective method for determining the sex of transgender patients which could help them in understanding their gender identity better and diagnose any underlying chromosomal aberration.
PubMed: 38304528
DOI: 10.4103/jomfp.jomfp_342_23 -
Journal of Oral and Maxillofacial... 2023Polymorphonuclear neutrophils are the most abundant leukocytes in humans and are key host cells in defence against invading microorganisms. The oral neutrophil count may...
BACKGROUND AND OBJECTIVES
Polymorphonuclear neutrophils are the most abundant leukocytes in humans and are key host cells in defence against invading microorganisms. The oral neutrophil count may be an indicator of the periodontal health status, which correlates with the severity of periodontal disease. This study attempts to quantify orogranulocytes utilising an oral rinse and to assess the usefulness of this method in evaluating the oral inflammatory load much the same way the circulating neutrophils are used to screen for patients with infection in extra-oral sites.
METHODS
A total of 125 participants were divided into five groups with 25 subjects in each group. The groups consisted of healthy, gingivitis, mild periodontitis, moderate periodontitis, and severe periodontitis. Participants were asked to rinse with 10 mL of 0.9% saline for 30 s and to expectorate. Samples were centrifuged at 2000 RPM for 10 min. The supernatant removed was suspended in 5 mL of Hanks's balanced salt solution. One millilitre of this suspension was mixed with 4 μL of acridine orange. A 10 μL aliquot of this suspension was then assessed on a haemocytometer, and the oral PMNs were counted using fluorescence microscopy.
RESULTS
The mean number of oral neutrophils (100,000 cells/mL) was the lowest in the healthy group and increased in ascending order across the different groups with the highest for severe periodontitis group.
CONCLUSION
The oral neutrophil counts increased with the severity of periodontal inflammation. This is an easy, safe, reliable, and non-invasive method of quantification of oral neutrophils.
PubMed: 38304508
DOI: 10.4103/jomfp.jomfp_418_21 -
Frontiers in Bioscience (Landmark... Jan 2024The present study aims to investigate the effect of Huaier on oxaliplatin (OXA) resistance in HCT-8 colorectal cancer (CRC) cells.
OBJECTIVE
The present study aims to investigate the effect of Huaier on oxaliplatin (OXA) resistance in HCT-8 colorectal cancer (CRC) cells.
METHODS
Oxaliplatin-resistant HCT-8/L CRC cells were used. The Cell Counting Kit-8, western blotting, quantitative real-time polymerase chain reaction, protein extraction kit, immunofluorescence and acridine orange staining assays were used in the study. The experiment results proved that Huaier has an influence on the Wnt/β-catenin signalling pathway, autophagy and drug resistance. The authors of the present study used chloroquine, an autophagy inhibitor and Wnt agonist 1 (a Wnt pathway agonist) to verify the present experiment.
RESULTS
The results showed that Huaier can regulate autophagy, inhibit the Wnt/β-catenin signalling pathway and reverse the drug resistance of OXA-resistant CRC cells.
CONCLUSIONS
This study proved that Huaier can regulate autophagy, inhibit the Wnt/β-catenin signalling pathway and reverse the drug resistance of OXA-resistant CRC cells.
Topics: Humans; Oxaliplatin; Wnt Signaling Pathway; beta Catenin; Colorectal Neoplasms; Cell Line, Tumor; Autophagy; Cell Proliferation; Complex Mixtures; Trametes
PubMed: 38287798
DOI: 10.31083/j.fbl2901015 -
Asian Pacific Journal of Cancer... Jan 2024Clerodendrum infortunatum L. has long been used in traditional medicine in Sri Lanka for tumours, cancer, and certain skin diseases. The present study aimed to assess...
OBJECTIVE
Clerodendrum infortunatum L. has long been used in traditional medicine in Sri Lanka for tumours, cancer, and certain skin diseases. The present study aimed to assess the anticancer properties of the aqueous extract of C. infortunatum L. root (AECIR) through the activation of the apoptotic pathway on hepatocellular carcinoma (HepG2) and thus give it a scientific validation. Further, the contribution of polyphenols in antioxidant activity and cell cytotoxicity was investigated.
METHODS
Powdered plant material was boiled with water (100°C) to obtained AECIR. The DPPH assay was used to determine the antioxidant potential. The activity of AECIR on HepG2 and normal rat fibroblast (CC1) cell growth was determined using MTT assay. The morphological changes related to apoptotic pathway was examined by Ethidium Bromide/Acridine Orange (EB/AO), Rhodamine 123 (Rh123) and DNA fragmentation assay.
RESULTS
The AECIR demonstrated antioxidant potential with an EC50 of 350.2 ± 1.5 ug/mL for DPPH assay. The HO•, H2O2 and •NO free radical scavenging activity was observed with EC50 of 19.7 ± 2.3, 11.7 ± 0.1 and 273.1 ± 0.9 ug/mL, respectively. The antiproliferative effect of AECIR on HepG2 cells was observed in a time and dose dependent manner with an EC50 of 239.1 ± 1.3 μg/mL while CC1 cells showed a nontoxic effect with an EC50 1062.7 ± 3.4 μg/mL after 24hrs treatment. A significant decrease in antioxidant activity (p<0.001) and 90% HepG2 cell viability was observed with polyphenol removed AECIR compared to the polyphenol present AECIR. The EB/AO uptake, depletion of mitochondrial transmembrane potential, and DNA fragmentation assay results revealed that the apoptosis was induced by AECIR.
CONCLUSION
The obtained result of the present study demonstrates that the antioxidant potential and antiproliferative activity of AECIR is attributed to the presence of polyphenols. Furthermore, the findings provide the scientific base for anti-cancer potential of AECIR.
Topics: Animals; Rats; Humans; Carcinoma, Hepatocellular; Polyphenols; Antioxidants; Hep G2 Cells; Clerodendrum; Hydrogen Peroxide; Plant Extracts; Liver Neoplasms; Cell Proliferation; Apoptosis
PubMed: 38285803
DOI: 10.31557/APJCP.2024.25.1.351 -
International Journal of Molecular... Jan 2024Furanocoumarins are naturally occurring compounds in the plant world, characterized by low molecular weight, simple chemical structure, and high solubility in most...
Furanocoumarins are naturally occurring compounds in the plant world, characterized by low molecular weight, simple chemical structure, and high solubility in most organic solvents. Additionally, they have a broad spectrum of activity, and their properties depend on the location and type of attached substituents. Therefore, the aim of our study was to investigate the anticancer activity of furanocoumarins (imperatorin, isoimperatorin, bergapten, and xanthotoxin) in relation to human glioblastoma multiforme (T98G) and anaplastic astrocytoma (MOGGCCM) cell lines. The tested compounds were used for the first time in combination with LY294002 (PI3K inhibitor) and sorafenib (Raf inhibitor). Apoptosis, autophagy, and necrosis were identified microscopically after straining with Hoechst 33342, acridine orange, and propidium iodide, respectively. The levels of caspase 3 and Beclin 1 were estimated by immunoblotting and for the blocking of Raf and PI3K kinases, the transfection with specific siRNA was used. The scratch test was used to assess the migration potential of glioma cells. Our studies showed that the anticancer activity of furanocoumarins strictly depended on the presence, type, and location of substituents. The obtained results suggest that achieving higher pro-apoptotic activity is determined by the presence of an isoprenyl moiety at the C8 position of the coumarin skeleton. In both anaplastic astrocytoma and glioblastoma, imperatorin was the most effective in induction apoptosis. Furthermore, the usage of imperatorin, alone and in combination with sorafenib or LY294002, decreased the migratory potential of MOGGCCM and T98G cells.
Topics: Humans; Sorafenib; Phosphatidylinositol 3-Kinases; Glioma; Furocoumarins; Glioblastoma; Astrocytoma; Chromones; Morpholines
PubMed: 38255833
DOI: 10.3390/ijms25020759 -
International Journal of Biological... Feb 2024Oral cancer incidence and mortality are increasing over time. The most common therapies for oral cancers are surgery and radiotherapy, either used alone or combined, and...
Oral cancer incidence and mortality are increasing over time. The most common therapies for oral cancers are surgery and radiotherapy, either used alone or combined, and immunotherapy can be also an option. Although there are several therapeutic options, none of them are completely effective, and in addition, there are numerous associated side effects. To overcome these limitations, researchers have been trying to reduce these drawbacks by using drug delivery systems that carry drugs for specific delivery to cancer cells. For that purpose, RNA-coated liposomes to selectively deliver the ligands C (acridine orange derivative) and dexamethasone to oral cancer cells were produced, characterized, and biologically evaluated. Firstly, the RNA structure and binding interaction with ligands (C and dexamethasone) were evaluated by circular dichroism (CD), thermal difference spectroscopy (TDS), nuclear magnetic resonance (NMR) and fluorescence titrations. The biophysical assays evidenced the formation of an RNA hairpin and duplex structure. Moreover, steady-state and time-resolved fluorescence intensity and anisotropy experiments show that C forms a complex with RNA and adopts an open conformation upon RNA binding. Then, RNA-coated liposomes were characterized by dynamic light scattering, and diameters near 160 nm were observed. Time-resolved anisotropy measurements of C loaded in RNA-functionalized liposomes indicate the co-existence of free C in solution (inside the liposome) and C bound to RNA at the external liposome surface. The RNA-functionalized liposomes loaded with C or dexamethasone mediated a significant reduction in the cell viability of malignant UPCI-SCC-154 cells while maintaining viable non-malignant NHDF cells. Additionally, the liposomes were able to internalize the cells, with higher uptake by the malignant cell line. Overall, the results obtained in this work can contribute to the development of new drug delivery systems based on RNA-coated liposomes.
Topics: Humans; Liposomes; Drug Delivery Systems; Cell Line; Mouth Neoplasms; Dexamethasone
PubMed: 38199539
DOI: 10.1016/j.ijbiomac.2023.129157