SCC-25 Cells Research Articles

Propranolol inhibits cell viability and expression of the pro-tumorigenic proteins Akt, NF-ĸB, and VEGF in oral squamous cell carcinoma

BackgroundPropranolol (PPL) has been suggested as an option for the treatment of various types of cancer. However, data regarding its effectiveness against oral cancer are scarce. Thus, we aimed to evaluate the antitumor potential of PPL in oral squamous cell carcinoma (OSCC) in vitro. MethodsOSCC cell lines, SCC-9, SCC-25, and Cal27, were treated with PPL at different times and concentrations. OSCC cells were treated with PPL alone or in combination with cisplatin (CDDP) or 5-fluorouracil (5-FU). Cell viability was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The expression of phosphorylated (p)-Akt, p-S6, p-PTEN, p-P65, and VEGF was verified by immunofluorescence. The migratory activity of OSCC cells was evaluated using a wound-healing assay. ResultsPPL reduced OSCC cell viability in a dose- and time-dependent manner. Concentrations above 300 µM, 110 µM, and 100 µM for SCC-9, Cal27, and SCC-25, respectively, significantly eliminated tumor cells. The combination of PPL with CDDP and 5-FU enhanced their antitumor effects. There was a modest difference between the use of the IC30 and IC50 of PPL in the combinatory options. PPL downregulated p-P65 NF-ĸB and VEGF expression in SCC-9 and Cal27 cells but not in SCC-25 cells. PPL inhibited the phosphorylation of Akt and s6 and increased the phosphorylation of PTEN in all OSCC cell lines studied. PPL inhibited OSCC cell migration after 24 h of treatment. ConclusionPPL was effective against oral cancer cells and enhanced standard-of-care. PPL inhibited cell viability and the expression of pAkt, NF-ĸB, and VEGF.

Photodynamic therapy mediated by nanoparticles Aluminum Chloro Phthalocyanine in oral squamous carcinoma cells.

The aim of this study is to investigate the antineoplastic potential of photodynamic therapy (PDT) mediated by an aluminum-phthalocyanine chloride nanoemulsion (AlPc-NE), against an oral squamous cell carcinoma (OSCC) cell line in vitro. Both OSCC (SCC9) and A431 cell lines were studied in vitro. Four study groups were used: Group 1 (phosphate-buffered saline [PBS]), Group 2 (PBS + 28.3J/cm2 irradiation), Group 3 (AlPc-NE alone), and Group 4 (AlPc-NE + 28.3J/cm2 irradiation). To test the effect of PDT with AlPc-NE, cell viability, migration, and cell death assays were performed. Moreover, the expressions of Ki-67 and TP53 were evaluated using immunoassays. The results showed that PDT mediated by all AlPc-NE concentrations evaluated (i.e., 0.7, 0.35, and 0.17nM AlPc) significantly reduced the viability of SCC9 cells. Migration and cell death assays also revealed that PDT with AlPc-NE significantly reduced the rate of migration and increased cell death compared to the control groups. In addition, it was found that PDT with AlPc-NE reduced Ki-67 and mutated TP53 immunoexpression. PDT with AlPc-NE is effective in reducing the viability and migration of SCC9. Moreover, PDT with AlPc-NE nanoemulsions reduces the cell proliferation and expression of mutant TP53.

In Vitro Assessment of the Cytotoxic and Antiproliferative Profile of Natural Preparations Containing Bergamot, Orange and Clove Essential Oils.

Medicinal plants and essential oils (EOs), in particular, were intensively studied in recent years as viable alternatives for antiproliferative chemical synthetic agents. In the same lines, the present study focuses on investigating the effects of natural preparations (emulsions) based on EOs obtained from Citrus bergamia Risso (bergamot-BEO), Citrus sinensis Osbeck (orange-OEO), and Syzygium aromaticum Merill et L. M. Perry (clove-CEO) on different healthy (human immortalized keratinocytes—HaCaT and primary human gingival fibroblasts—HGF) and human tumor cell lines (human melanoma—A375 and oral squamous carcinoma—SCC-4) in terms of the cells’ viability and cellular morphology. The obtained results indicate that the CEO emulsion (ECEO) induced a dose-dependent cytotoxic in both healthy (HaCaT and HGF) and tumor (A375 and SCC-4) cells. OEO emulsion (EOEO) increased cell viability percentage both for HaCaT and A375 cells and had an antiproliferative effect at the highest concentration in HGF and SCC-4 cells. BEO emulsion (EBEO) decreased the viability percentage of SCC-4 tumor cells. By associating OEO with CEO as a binary mixture in an emulsified formulation, the inhibition of tumor cell viability increases. The E(BEO/OEO) binary emulsion induced an antiproliferative effect on oral health and tumor cells, with a minimal effect on skin cells. The non-invasive tests performed to verify the safety of the test compound’s emulsions at skin level indicated that these compounds do not significantly modify the physiological skin parameters and can be considered safe for human skin.

Mechanism of 5-fluorouracil nanoparticles on releasing skin squamous cell carcinoma through regulation of Wnt/β-catenin expression

5-FU nanoparticles were prepared in this study to discuss mechanism of 5-fluorouracil nanoparticles on releasing SSCC through regulating Wnt/β-catenin expression. The characteristics of nanomaterial were analyzed through electron microscope. The releasing characteristics in vitro and in vivo were also detected. Proliferation and apoptosis of SCC cell line in control and 5-FU nanoparticles groups were detected through FCM and MTT method. The tumor volume was recorded and average particle size was 310 nm with uni-form distribution. The apoptosis of MCF-7 cells in the 5-FU nanoparticles group was increased notably, and there were abundant circular floating apoptotic cells. The quantity of active MCF-7 cells was reduced notably and proliferation of SSCC could be restrained by 5-FU nanoparticles through regulation of Wnt/β-catenin expression.

Synthesis of Biotinylated PAMAM G3 Dendrimers Substituted with R-Glycidol and Celecoxib/Simvastatin as Repurposed Drugs and Evaluation of Their Increased Additive Cytotoxicity for Cancer Cell Lines.

Simple SummaryNew anti-cancer drugs can be created through the combination of well-known substances acting on different molecular targets. Repurposed drugs selected in this way can then be attached to special carriers that have specific, high affinity for cancer cells. In this study, two anti-glioma drugs, celecoxib and simvastatin, were chosen and linked to the PAMAM G3 dendrimer targeted to cancer cells by attached biotin and R-glycidol. In vitro studies performed with human glioblastoma (U-118 MG) and squamous cell carcinoma (SCC-15) cells revealed that dendrimer conjugate containing both celecoxib and simvastatin was 20–50 times more potent than either drug administered alone or in combination. Thus, the obtained combined conjugate can be considered as a potential candidate for a new therapy of malignant glioblastoma.Recent achievement in anticancer therapy considers the application of repurposed drugs in optimal combinations with the use of specific carriers for their targeted delivery. As a result, new optimized medications with reduced side effects can be obtained. In this study, two known anticancer drugs, celecoxib and/or simvastatin, were conjugated covalently with PAMAM G3 dendrimer and tested in vitro against human squamous carcinoma (SCC-15-15) and glioblastoma (U-118 MG) cells, as well as normal human fibroblasts (BJ). The obtained conjugates were also substituted with biotin and R-glycidol to increase their affinity for cancer cells and were characterized with NMR spectroscopy and dynamic light scattering technique. Conjugates furnished with two celecoxib and four simvastatin residues revealed the very high effectiveness and dramatically decreased the SCC-15 and U-118 MG cell viability at very low concentrations with IC50 equal to about 3 µM. Its action was 20–50-fold stronger than that of either drug alone or as a mixture. Combined conjugate revealed also additive action since it was 2–8-fold more effective than conjugates with either single drug. The combined conjugate revealed rather low specificity since it was also highly cytotoxic for BJ cells. Despite this, it may be concluded that biotinylated and R-glycidylated PAMAM G3 dendrimers substituted with both celecoxib and simvastatin can be considered as a new perspective anticancer agent, effective in therapy of malignant, incurable glioblastomas.

HR-LCMS and In vitro cytotoxicity analysis of Alseodaphne semecarpifolia stem bark and leaf methanol extracts

A. semecarpifolia (Lauraceae) has been employed by Indian traditional healers to treat various human cancers. The cytotoxic property of A. semecarpifolia was evaluated by extracting secondary metabolites from stem bark and leaves. The stem bark methanol extract (SBME) and leaf methanol extract (LME) were subjected to HR-LCMS analysis, followed by evaluating their cytotoxic potential against MCF-7, SCC-9, HeLa, A-549 and L6 cells by MTT assay. The phytochemical screening of SBME and LME revealed the presence of alkaloids, flavonoids, terpenoids, steroids, saponins, phenolics and glycosides. HR-LCMS analysis of SBME and LME revealed the presence of 31 and 28 active compounds respectively. Among them Cucurbitacin A and Gambogic acid are the two compounds identified with anticancer properties. SBME showed potent cytotoxicity on MCF-7 and SCC-9 cells, whereas LME showed significant cytotoxicity only on MCF-7 cells. Vinblastine sulphate was used as a reference standard, even though Vinblastine is an efficient anticancer drug it affected the viability of normal cells. In comparison with Vinblastine, both SBME and LME showed very less toxicity on normal cells. Hence, the present study suggested that stem bark and leaves of A. semecarpifolia are the possible chemotherapeutic agents having potential cytotoxic activity against MCF-7 and SCC-9 cells.

MiR-34c-3p upregulates erastin-induced ferroptosis to inhibit proliferation in oral squamous cell carcinomas by targeting SLC7A11

BackgroundMiRNA is a small molecule RNA that plays an important role in a variety of physiological and pathological processes., and miR-34c-3p has been demonstrated to be closely related to the occurrence of tumors. Ferroptosis is a new form of cell death characterized by lipid-based reactive oxygen species accumulation. However, it is still unclear how miR-34c-3p influences the development of oral squamous cell carcinoma (OSCC) by regulating ferroptosis. Therefore, the main objective of this study was to explore the role and mechanism of miR-34c-3p in OSCC. Materials and methodsThe expression of miR-34c-3p in OSCC and matched normal tissues was detected by quantitative real-time PCR (qRT-PCR). Subsequently, the effect of miR-34c-3p overexpression on cell proliferation and ferroptosis was evaluated using CCK8, colony formation assays, Live/Dead staining, Western blotting analysis, ROS, MDA, and GSH assay. ResultsThe results showed lower expression of miR-34c-3p in OSSC compared with normal tissues. Overexpression of miR-34c-3p in SCC-25 cells suppressed cell proliferation. In addition, the overexpression of miR-34c-3p promoted ferroptosis by increasing ROS, MDA, and iron and decreasing GSH and GPX4 levels in SCC-25 cells. ConclusionOur findings revealed a novel strategy to upregulate erastin-induced ferroptosis in OSCC through the miR-34c-3p/SLC7A11 axis, suggesting new insights into OSCC and a potentially useful therapeutic strategy for OSCC.

Evaluation of Curcumin Therapeutic Effects on Histological Subtypes of Canine Mammary Gland Tumours

Canine mammary gland tumors (CMGTs) are the most frequent types of cancer in bitches and proposed as a model of human breast cancer. The anticancer effect of curcumin on human breast cancer has been extensively studied. The aim of this study was to evaluate the therapeutic effect of curcumin in two different histologies (simple carcinoma [SC] and squamous cell carcinoma [SCC]) of CMGTs. Primary canine mammary cells were isolated from the tumoral tissues surgically resected from two bitches (Case 1 and Case 2). Cell viability, apoptotic percentage, cell cycle progression and the changes in the cell morphology were evaluated. Curcumin inhibited the growth of both SC (Case 1) and SCC (Case 2) cells. However, Case 1 cells (43.48% ± 3.87% at 0.5 µM) were more sensitive to curcumin than Case 2 cells (59.36% ± 2.09% at 0.5 µM). Curcumin induced total apoptotic cell death through G0/G1 arrest, and this effect was more profound in Case 1 cells. Furthermore, cytoplasmic vacuolization, apoptotic bodies and membrane blebbing were observed in both cells following curcumin treatment. Our findings provide a novel approach for the effects of curcumin as a natural compound on CMGTs. Further investigations should be performed to investigate the molecular mechanisms of the differences in curcumin efficacy for different histological subtypes.

Antibacterial, Cytotoxic, and Cellular Mechanisms of Green Synthesized Silver Nanoparticles against Some Cariogenic Bacteria (Streptococcus mutans and Actinomyces viscosus)

Background. The present study focused on the green synthesis of silver nanoparticles (AgNPs) using the Astragalus spinosus Forssk. aqueous extract. In addition, we evaluated the antibacterial activity of AgNPs as well as some cellular mechanisms against Actinomyces viscosus and Streptococcus mutans as the most causative agents of tooth decay. Methods. In this study, AgNPs were green synthesized by the precipitation method based on the reduction of silver ions (AgNO3) by A. spinosus extract. Antibacterial effects of the green synthesized AgNPs were performed by measuring the minimum inhibitory concentration (MIC) and minimum bactericidal concentrations (MBC) through micro broth dilution method. In addition, we evaluated the reactive oxygen species (ROS) production, nucleic acid leakage, and protein leakage as the main antibacterial mechanisms of the green synthesized AgNPs against A. viscosus and S. mutans. The cytotoxicity effects of AgNPs against on human normal (NOF18 cells) and oral cancer (SCC4 cells) cell lines were also evaluated using MTT assay. Results. The green synthesized AgNPs have a spherical shape and are relatively uniform in size in the range of 30‐40 nm. The MIC values for S. mutans and A. viscosus of the green synthesized AgNPs were 10.6 and 13.3 μg/ml, respectively, whereas the MBC values for S. mutans and A. viscosus of the green synthesized AgNPs were 21.3 and 26.6 μg/ml. The findings exhibited that ROS production, nucleic acid leakage, and protein leakage were increased after treatment of A. viscosus and S. mutans by the green synthesized AgNPs. The results demonstrated that the 50% inhibitory concertation (IC50) values of AgNPs on NOF18 and SCC4 cells were 93.3 μg/ml and 41.2 μg/ml, respectively. Conclusion. Overall, the results of this study showed that A. spinosus extract has a good ability to produce silver nanoparticles. The AgNPs produced have significant antibacterial effects against some tooth decay bacteria. Our results also revealed that the green synthesized AgNPs are more cytotoxic against cancerous cell line than normal cell line. Further in vivo studies are required to investigate the side effects and to evaluate the effectiveness of these bacteria.

The role of long non-coding RNA LINC01296 in oral squamous cell carcinoma: a study based on bioinformatics analysis and in vitro validation.

Background: Oral squamous cell carcinoma (OSCC) is a common malignancy in the oral cavity that represents a significant global health problem. Multivariate analysis has shown that long non-coding RNA LINC01296 plays an important role in cancer biology. However, the functions of LINC01296 in OSCC are still unknown.Methods: The RNA profiles and clinicopathological information of OSCC patients and healthy subjects were downloaded. The expression level and prognostic value of LINC01296 were assessed. The functions and pathways of LINC01296were explored using the Gene Set Enrichment Analysis (GSEA) and functional analysis. The expression of LINC01296 in OSCC tissues and cell lines was determined by RT-qPCR. MTS assay was used to evaluate cell growth. The migration and invasion capacities of cells were assessed by wound healing assay and Transwell assay.Results: LINC01296 was overexpressed in the TCGA-OSCC datasets. High LINC01296expression was strongly correlated with poor outcomes of OSCC patients. LINC01296 was overexpressed in OSCC tissues compared with para-carcinoma tissues. Moreover, the expression of linc01296 was higher in CAL-27, HSC-2, and SCC-25 cells than in normal human oral keratinocytes (NHOKs). Functional analysis suggested that LINC01296might be involved in the regulation of the Wnt and MAPK signaling pathways. Additionally, LINC01296 deficiency suppressed the growth, migration, and invasion of OSCC cells, whereas overexpression of TFAP2A-AS1 cause opposite results.Conclusion: Our study showed that LINC01296 promoted OSCC cell growth, migration, and invasion, suggesting that LINC01296 might be a potential therapeutic target for OSCC.

Differential Expression of Cellular and Exosomal MicroRNA Isolated from Oral Cancer Cells and their Resistance to Chemotherapy

MicroRNAs are abundant small non-coding RNA with a variety of known functions, including transcriptional activation and inhibition. Recent evidence has suggested that microRNA expression may influence the responsiveness of some cancers to chemotherapy, including liver and lung cancers. Some evidence has now suggested that specific microRNAs, such as miR-21, miR-155, and miR-375, may influence oral cancer responsiveness to chemotherapy – although much remains to be discovered. Based on the lack of evidence in this area, the primary objective of this study was to evaluate microRNA expression and responsiveness among oral cancers. Five commercially available oral cancer cell lines (SCC4, SCC9, SCC15, SCC25, CAL27) were obtained from ATCC and cultured and chemotherapy resistance to Cisplatin, Fluorouracil or 5-FU and Paclitaxel or Taxol was assessed. Exosomes were then isolated, confirmed, and processed using Particle Metrix Nano Tracking Analysis (NTA) Subsequently, RNA was isolated from both the exosomes and cellular fractions and qPCR screening was performed to determine the expression of microRNA from cellular and exosomal isolates. Growth assays revealed that SCC15 assays were the least resistant, while CAL27 and SCC4 cells exhibited moderate resistance, and SCC9 and SCC25 cells exhibited strong and differential resistance to these chemotherapeutic agents. The screening revealed all cancers expressed miR-21 and miR-133 with differential expressions of miR-27, miR-135, miR-155, and miR-375 observed. However, the most resistant cell lines, SCC9 and SCC25, were the only cells to express miR-375 and were also the only cells that did not express miR-27, suggesting an association between chemotherapeutic resistance and expression of these specific microRNAs. In addition, miR-21 and miR-133 were identified in exosomes from all oral cancers with differential results observed with miR-133 and miR-135-although miR-27 and miR-375 were not found among any exosomes. Although much remains to be elucidated about the functional roles of these differentially expressed microRNAs, the findings of this study suggest that specific microRNAs including miR-27 and miR-375 may, in fact, function in distinct, different, and opposite pathways among these cell lines. Future research endeavors will need to evaluate the potential role of these microRNAs not only to validate their predictive capabilities as biomarkers but also to ascertain which functional pathways may be involved in the development and progression of oral cancers.

Down-regulation of AMPD3 Is Associated With Poor Survival in Head and Neck Squamous Cell Carcinoma.

Adenosine monophosphate deaminase 3 (AMPD3) is an isoenzyme involved in the regulation of the energetic metabolism of mammalian cells. Cancer cells have a high demand for their energy supply. This experimental study aimed to illustrate the role of AMPD3 in human head and neck squamous cell carcinoma (HNSCC). Real-time quantitative reverse transcription-polymerase chain reaction was used to investigate the expression of the AMPD3 gene in human HNSCC tissues to assess the changes in cancerous and noncancerous parts and the correlation with different tumor behavior. The functions of AMPD3 were investigated using wound-healing and migration assays. AMPD3 was significantly down-regulated in cancerous tissues of HNSCC (p=0.001) and this was correlated with more advanced tumor and clinical stages. Patients with high expression had better 5-year survival. AMPD3 knock-down in SCC-4 and SCC-25 cells demonstrated reduction of proliferation but increased migration and invasion. To our knowledge, this is the first report evidencing the expression pattern of AMPD3 in HNSCC and demonstrated that high AMPD3 expression might represent a good prognostic biomarker. AMPD3 may have an antiproliferative potential but its down-regulation may not contribute to reducing the migration and invasion of HNSCC cells.

Evaluation of cytotoxic effects of fungal origin nanosilver particles on oral cancer cell lines: An in vitro study.

Oral cancer is often associated with poor prognosis and it is found that conventional treatment options cause severe side effects, adjacent tissue disfigurement, and loss of function. Recently, silver nanoparticles (AgNPs) paved their path for cancer treatment. This study aimed to investigate cytotoxic effects of fungal procured AgNPs on oral squamous cell carcinoma (SCC-9) cell line using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The silver nanoparticles were biosynthesized using the fungi Fusarium semitectum. Cell lines were cultured in a 1:1 ratio of Dulbecco's Modified Eagle's medium and Ham's F12 medium and subcultured in a T-75 cm2 flask. Cell count was adjusted to 1 × 105 cells/ml; 50,000 cells/well were seeded into a 96 well plate and incubated at 37°C, for 24 h in 5% CO2 humidified conditions. AgNPs (1.75-50 μl/ml) were added to the plates and further incubated at 37°C for another 24 h. Medium containing cells without AgNPs were used as a control group. Later, 20 μl of MTT was added to each well and incubated for 6 h at above-mentioned conditions. About 0.1 mL of Dulbecco's Modified Eagle's Medium solution was added to each well to solubilize formazan. The absorbance was measured using a Tecan reader at 540 nm. The experiment was repeated thrice independently. The percentage (%) inhibition of growth and the AgNP's concentration that prevents the cell growth by 50% (IC50) were determined. Significant dose-dependent inhibition of the growth of SCC-9 cell lines was seen and IC50 was found at 12 μl/ml concentration of AgNPs. Biosynthesized AgNPs of fungal origin exhibit effective anticancer properties against the SCC-9 cell line.

Ginsenoside Rd inhibits migration and invasion of tongue cancer cells through H19/miR-675-5p/CDH1 axis.

Objective Tongue squamous cell carcinoma (TSCC) is an oral cancer, with high malignancy and frequent early migration and invasion. Only a few drugs can treat tongue cancer. Ginsenoside Rd is a ginseng extract with anti-cancer effects. Many noncoding RNAs are abnormally expressed in tongue cancer, thus influencing its occurrence and development. H19 and miR-675-5p can promote cancer cell growth. This study aimed to analyze the regulation effect of ginsenoside Rd on H19 and miR-675-5p in tongue cancer.Methodology We used CCK8 and flow cytometry to study the growth and apoptosis. Transwell assay was used to assess invasion; wound-healing assay to assess migration; and colony formation assays to test the ability of cells to form colonies. H19, miR-675-5p, and CDH1 expressions were analyzed by qPCR. E-cadherin expression was detected using western blot. CRISPR/cas9 system was used for CDH1 knockout.Results Ginsenoside Rd inhibited the growth and increased the apoptosis of SCC9 cells. Ginsenoside Rd also inhibited the migration and invasion of SCC9 cells. H19 and miR-675-5p were highly expressed, while CDH1 and E-cadherin expressions were low. H19 and miR-675-5p promoted SCC9 metastasis. In contrast, CDH1 and E-cadherin inhibited the metastasis of SCC9 cells. Bioinformatics analysis showed that miR-675-5p was associated with CDH1. H19 and miR-675-5p expressions decreased after ginsenoside Rd treatment, while CDH1 and E-cadherin expressions increased.Conclusions Ginsenoside Rd inhibits tongue cancer cell migration and invasion via the H19/miR-675-5p/CDH1 axis.

In vitro cytotoxic activity using fruits of Annona muricata and aerial parts of Euphorbia tirucalli against lung and oral cancer cell lines

Aim: Cancer is one of the life-threatening diseases which cause severe pathological conditions, leading to mortality. The exhaustive data on the vital role of medicinal plants in combating cancer and related diseases are available since antediluvian times. Materials and Methods: In this present study, cell viability, morphological changes, and IC50 were evaluated by in vitro 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltertazoliumbromide) assay. Hydro-alcoholic and ethyl acetate extracts of fruits Annona muricata (AM) and aerial parts Euphorbia tirucalli (ET) against lung (A549, H1975) and oral (SCC9, SCC25) were used to investigate the potential antitumor activity. Results: Ethyl-acetate extract of AM and ET showed the highest IC50 value, 89.48 μg/mL and 119.2 μg/mL against lung cancer cell line A-549. Among four study extracts, the IC50 value of hydroalcoholic extract of AM showed 184.3 μg/mL against H1975 cell line. The hydroalcoholic extract of AM and ethyl acetate extract of EA showed the IC50 value of 149.7 μg/mL and 156.2 μg/mL against SCC9 cell line. Hydroalcoholic and ethylacetate extract of ET showed the highest cytotoxic potency against SCC25 139.2 μg/mL and 205.6 μg/mL compared to AM. Further, all the study extracts exhibited decrease in % cell viability in the dose-dependent manner. Conclusion: This comprehensive data obtained from the in vitro study indicate that the extracts that showed toxicity toward cancer cell lines can be considered potential chemotherapeutic agents. The data suggest to further confirm the anticancer potential by conducting in vitro and in vivo studies, so that the study plants may be adopted in the treatment of lung and oral cancer.

Design, synthesis, cytotoxic, and anti-inflammatory activities of some novel analogues of aloe-emodin isolated from the rhizomes of Rheum emodi

In connection with our continuous efforts in the synthesis of derivatives from major compounds isolated from traditional medicinal plants, in the present study we have attempted to synthesize the furan-conjugated aloe-emodin derivatives (5a–j) using a three-component reaction. The synthesized derivatives were assessed for anticancer activity against five different cancer cell lines using the in vitro MTT assay and the results showed that most of the derivatives are potent against cancer cells comparing with the control. Compounds 5a and 5e showed excellent activity against all the cancer cells with less than 12.5 µM and arrested the cell cycle at the G0/G1 phase in both CAL27 and SCC9 cells. Compound 5e induces the early apoptosis in CAL27 cells and compounds 5a and 5e induce early and late apoptosis, respectively, in SCC9 cells. Moreover, compounds 5b, 5c, 5i, and 5j showed excellent anti-inflammatory activity in LPS-stimulated RAW 264.7 cells by inhibiting IL-6 production. The molecular docking studies revealed that compound 5e has strong interaction with the CLK kinase and protein kinase II through hydrogen binding Asp325 and Lys290.

Pharmacological fatty acid synthase inhibitors differently affect the malignant phenotype of oral cancer cells.

ObjectiveFatty acid synthase levels are associated with aggressiveness, prognosis, and risk of metastasis in oral squamous cell carcinomas. This enzyme contains seven catalytic domains and its inhibition by synthetic or natural drugs has antineoplastic properties such as C75, which is a synthetic inhibitor of the β- ketoacyl synthase domain, the antibiotic triclosan, ligand of the enoyl reductase domain, and the antiobesity drug orlistat, which inhibits the thioesterase domain. Here, we sought to investigate and compare the in vitro effects of C75, triclosan, and orlistat on malignant phenotypes of the cell line SCC-9: proliferation, cell cycle, apoptosis, adhesion, migration, and invasion. DesignHalf-maximal inhibitory concentration (IC50) was determined using cell viability assays. Cell death and cell cycle progression were analyzed by Annexin V-PE/7-ADD-PerCP labeling and propidium iodide staining, respectively. Cell migration and invasion were assayed by transwells assays and cell adhesion using collagen and fibronectin. ResultsC75 showed the lowest IC50 and higher inhibition of lipid droplets at low concentrations and reduced cell motility. Triclosan showed the intermediate IC50 value, excellent reduction of lipid bodies at the IC50 when compared with C75 and orlistat. Also, triclosan reduced cell cycle progression, adhesion, migration, and invasion of SCC-9 and induced the highest levels of apoptosis. Orlistat promoted cell cycle arrest, but showed the lowest induction of apoptosis and did not affected invasion and adhesion of SCC-9. ConclusionAltogether, despite the particular effects of the analyzed fatty acid synthase inhibitors, triclosan showed to better interfere in tumorigenic phenotypes of SCC-9 cells.

PKMYT1 regulates the proliferation and epithelial-mesenchymal transition of oral squamous cell carcinoma cells by targeting CCNA2.

Oral squamous cell carcinoma (OSCC) has gradually become a global public health issue in recent years. Therefore, the current study aimed to explore the mechanism of OSCC development and to identify a potential target that may be used in its treatment. The expression of protein kinase, membrane-associated tyrosine/threonine 1 (PKMYT1) and cyclin A2 (CCNA2) in SCC-9 cells was determined prior to and following transfection with short hairpin RNA targeting PKMYT1. Cell proliferation, colony-forming ability, migration and invasion were determined using Cell Counting Kit-8, colony formation, wound healing and Transwell assays, respectively. Furthermore, the expression of epithelial-mesenchymal transition (EMT)- and migration-related proteins were evaluated using western blot analysis. Additionally, co-immunoprecipitation was used to verify the binding of PKMYT1 and CCNA2. The results revealed that PKMYT1 was highly expressed in OSCC cells and that PKMYT1 knockdown could inhibit proliferation, colony formation, migration, invasion, EMT and CCNA2 expression in SCC-9 cells. In addition, PKMYT1 was demonstrated to bind to CCNA2, and knocking down PKMYT1 resulted in inhibitory effects on cell proliferation, colony formation ability, migration, invasion and EMT by downregulating CCNA2 expression. PKMYT1 was observed to regulate the proliferation, migration and EMT of OSCC cells by targeting CCNA2, which may be used in the future to improve OSCC treatment.

Photothermal effect of nano-copper sulfide against tongue squamous cell carcinoma

To investigate the anti-tumor effect of BSA@CuS-PEG nanocomposites on tongue squamous cell carcinoma. Transmission electron microscopy, dynamic light scattering, Zeta potential and ultraviolet absorption spectroscopy were used to characterize the synthesized BSA@CuS-PEG nanocomposite, whose photothermal properties was assessed with near infrared Ⅱ region excitation light (1064 nm). The cytotoxicity of the nanocomposite in Cal27 and SCC9 cells was evaluated using CCK-8 assay, and its effect on cell cycle distribution was analyzed using flow cytometry. The in vivo antitumor effect of BSA@CuS-PEG was investigated in a Balb/c mouse model bearing subcutaneous Cal27 tumor xenograft. The synthesized BSA@CuS-PEG nanocomposite showed a temperature variation (ΔT) of about 30 ℃ under near-infrared (NIR) irradiation (0.5 W/cm2), suggesting its excellent photothermal sensitivity. CCK-8 assay showed that BSA@CuS-PEG had no significant toxicity to tumor cells, but upon NIR irradiation, the nanocomposite produced a significant stronger inhibitory effect on Cal27 and SCC9 cells than free nanocomposites (P < 0.001). Cell cycle analysis showed that compared with free nanocomposites, BSA@CuS-PEG plus NIR irradiation caused more obvious cell cycle arrest at G2/M phase in tongue cancer cells (P < 0.001). In the tumor-bearing mice, BSA@CuS-PEG combined with NIR irradiation produced a significant anti-tumor effect as compared with saline treatment plus NIR irradiation (P < 0.001). The BSA@CuS-PEG nanocomposite shows prominent photothermal properties and good anti-tumor effects both in vivo and in vitro, and thus provides a promising method for non-invasive early diagnosis and non-surgical treatment of primary tongue cancer.

MicroRNA-573 inhibits cell proliferation, migration and invasion and is downregulated by PICSAR in cutaneous squamous cell carcinoma.

The incidence of cutaneous squamous cell carcinoma (cSCC) has been increasing in recent years. Meanwhile, microRNAs have been found to play vital roles in various cancers, including cSCC. This study aimed to investigate the expression of microRNA-573 (miR-573) in cSCC, its relationship with long non-coding RNA PICSAR and analyze its biological role. The relationship between PICSAR and miR-573 was confirmed by dual-luciferase reporter assay and Pearson’s correlation coefficient analysis. The levels of PICSAR and miR-573 were measured using quantitative Real-Time polymerase chain reaction. Cell Counting Kit-8 assay was used to evaluate the cSCC cell proliferation ability. The migration and invasion abilities of cSCC cells were evaluated by Transwell assay. PICSAR expression was increased and miR-573 was decreased in tumor tissues and cSCC cell lines. PICSAR and miR-573 can bind directly, and miR-573 expression was downregulated by PICSAR in cSCC. Overexpression of miR-573 significantly inhibited the proliferation, migration, and invasion abilities of A431 and SCC13 cells. In addition, miR-573 overexpression reversed the promotion effects of PICSAR overexpression on cSCC cell proliferation, migration, and invasion abilities. In conclusion, our findings indicated that miR-573 expression was decreased in tumor tissues and cSCC cells and was downregulated by PICSAR in cSCC. Additionally, miR-573 overexpression inhibited cSCC cell proliferation, migration and invasion, and reversed the promotion effects of PICSAR overexpression on cSCC cell biological functions. Thus, miR-573 might function as a tumor suppressor and might be involved in the regulatory effects of PICSAR on tumorigenesis in cSCC.