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Background: Moringa peregrina Forssk is a well-known plant in ethnomedicine due to its widespread uses in various diseases like cough, wound healing, rhinitis, fever, and detoxification. The plant seeds contain compounds that are cytotoxic to many cancer cells. During the therapeutic use of plants via the oral route, some compounds present in the plants may be cytotoxic to normal cell lines and red blood cells. Objective: This study was the first report of investigation of the cytotoxic profile on oral cancer, CAL 27, cell line, and hemolytic activities on human erythrocytes of Moringa peregrina seeds ethanolic extract (MPSE). Methods: MPSE was screened for its cytotoxic effect against oral cancer, CAL 27, cell line using 3-(4, 5-dimethylthiazol-2-yl)-2, 5,-diphenyltetrazolium bromide (MTT) assay. The toxicity of MPSE on human erythrocytes was determined by in vitro hemolytic assay. Results: MPSE showed significant anti-proliferative activity against oral cancer, CAL 27 cell line at lower concentrations with half maximal inhibitory concentration (IC50) value of 21.03 µg/mL. At 1,000 µg/ml of MPSE, the maximum hemolysis was found to be 14.3% which is within safer limit. Conclusions: This study revealed a potential anti-oral cancer of MPSE and provided a baseline for its potential use in oral cancer treatment with minimum hemolytic effect on human RBCs.
La Moringa peregrina Forssk es una planta muy conocida en etnomedicina debido a sus usos generalizados en diversas enfermedades como la tos, la cicatrización de heridas, la rinitis, la fiebre y la desintoxicación. Las semillas de la planta contienen compuestos citotóxicos para muchas células cancerosas. Durante el uso terapéutico de las plantas por vía oral, algunos compuestos presentes en ellas pueden ser citotóxicos para las líneas celulares normales y los glóbulos rojos. Objetivo: Este estudio fue el primer informe de investigación del perfil citotóxico sobre el cáncer oral, CAL 27, línea celular, y las actividades hemolíticas en eritrocitos humanos del extracto etanólico de semillas de Moringa peregrina (MPSE). Métodos: Se examinó el efecto citotóxico del MPSE contra la línea celular de cáncer oral CAL 27 mediante el ensayo con bromuro de 3-(4, 5-dimetiltiazol-2-il)-2, 5,-difeniltetrazolio (MTT). La toxicidad del MPSE sobre los eritrocitos humanos se determinó mediante un ensayo hemolítico in vitro. Resultados: MPSE mostró una actividad antiproliferativa significativa contra el cáncer oral, línea celular CAL 27 a concentraciones más bajas con un valor de concentración inhibitoria media máxima (IC50) de 21,03 µg/mL. A 1.000 µg/ml de MPSE, la hemólisis máxima fue del 14,3%, lo que está dentro del límite de seguridad. Conclusiones: Este estudio reveló un potencial anticancerígeno oral de MPSE y proporcionó una base para su uso potencial en el tratamiento del cáncer oral con un efecto hemolítico mínimo en los glóbulos rojos humanos.
Assuntos
Humanos , Moringa , Neoplasias Bucais , Citotoxinas , Eritrócitos , Medicina TradicionalRESUMO
@#Objective: To study the effects of vitamin C (VC) on reversing cisplatin (DDP) resistance in oral squamous cell carcinoma (OSCC) and the mechanism. Methods: Human OSCC CAL27 cells were cultured in vitro and DDP-resistant CAL27 cell line (CAL27/ DDP) was screened by increasing concentration gradient method. Plate clone formation assay, CCK-8, Wound healing assay, Annexin V-FITC/PI staining flow cytometry were used to determine the effects of DDP alone or in combination with VC on colony formation, proliferation, migration and apoptosis of CAL27/DDP cells. Western blotting was used to detect the expression level of P-gp protein in CAL27 cells, CAL27/DDP cells and VC treated CAL27/DDP cells. Results: The inhibition concentration (IC50) of DDP increased significantly in CAL27/DDP cells as compared with that in CAL27 cells (P<0.05), indicating CAL27/DDP was DDP-resistant.After the combination with VC, the IC50 of DDP on CAL27/DDP cells was significantly reduced compared with that of DDP alone (P<0.05). DDP combined with VC significantly inhibited the migration of CAL27/DDP cells (P<0.01), and promoted the apoptosis rate (P<0.01). The expression level of P-gp protein in CAL27/DDP cells was increased compared with that in CAL27 cells (P<0.05), but decreased after VC intervention (P<0.05). Conclusion: VC can reverse DDP-resistance in OSCC cells by inhibiting P-gp protein expression.
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@# Objective: To investigate the molecular mechanisms of lncRNA XIST/miR-34a-5p/SIRT6 axis regulating the proliferation and metastasis of oral squamous cell carcinoma (OSCC) cells. Methods: Specimens of tumor tissues and paracancer tissues of 47 patients with OSCC, who visited the Qingdao Stomatological Hospital from March 2013 to March 2018, were enrolled in this study. qPCR was performed to measure the mRNA expressions of lncRNA XIST, miR-34a-5p and SIRT6 in OSCC tissues and cell lines. WB was performed to measure the protein levels of SIRT6, Ki67, pcDNA, cleaved-caspase3, cleaved-caspase8, E-cadherin and vimentin in OSCC tissues and cell lines. CCK-8 assay was performed to observe the effect of lncRNA XIST knockdown on proliferation of Cal-27 and Tca-8113 cells; Tanswell assay was performed to detect migration and invasion of Cal-27 and Tca-8113 cells; flow cytometry was performed to detect the apoptosis of Cal-27 and Tca-8113 cells; and dual luciferase reporter assay was performed to verify the relationship between lncRNAXIST and miR-34a, or miR-34a and SIRT6. Results: Expressions of lncRNAXIST and SIRT6 were up-regulated in OSCC tissues and cell lines (all P<0.05), reversely, miR-34a-5p was down-regulated in OSCC tissues and cell lines (P<0.01). lncRNA XIST knockdown significantly suppressed OSCC cells proliferation, migration and invasion, and induced apoptosis of OSCC cells (all P<0.01); however, simultaneous transfection with miR-34a-5p inhibitor or pcDNA-SIRT6 vector exerted opposite effect. lncRNA XIST knockdown significantly increased cell proliferation and metastasis related protein expression in OSCC cells (all P< 0.01), and simultaneous transfection with miR-34a-5p inhibitor or pcDNA-SIRT6 vector exerted opposite effect. In addition, lncRNA XIST directly targeted miR-34a-5p, and miR-34a-5p targeted SIRT6, lncRNA XIST upregulated SIRT6 expression via inhibiting miR34a-5p (P<0.01). Conclusion: Taken together, lncRNA XIST/miR-34a-5p/SIRT6 axis regulates the proliferation and metastasis of OSCC cells and provides potential therapeutic targets for OSCC.
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Objective @#To investigate the expression of targeting protein for xenopus kinesin-like protein2 (TPX2) in tongue squamous cell carcinoma (TSCC) tissues and explore its effect on cell proliferation and apoptosis.@*Methods@#30 cases of TSCC tissues, paired normal tissues were collected in Dongguan People's Hospital during 2013-2016. The mRNA and protein expression level of TPX2 was determined by qRT-PCR and western blot, respectively and analyzed The correlation of TPX2 expression level and clinic opathological parameters. Cal27 cell was transfected with siRNAs to knockdown the expression of TPX2, then cell proliferation, cell apoptosis and related proteins (cleaved caspase 3 and caspase 3) were detected by MTT assay, flow cytometry and western blot. @*Results@#TPX2 was highly expressed in (t=3.254, P=0.002 9) tumor tissues at mRNA level compared to adjacent normal parts. In protein level, TPX2 was highly expressed (66.7%) in tumor tissues, TPX2 expression level of 20 case was higher than the corresponding tissue adjacent to carcinoma (20/30, t=2.862, P=2.862), and high expression of TPX2 was related to T staging, lymph node metastasis of tongue cancer. Knockdown TPX2 effectively reduced cell proliferation, increased apoptosis rate (F=342.9, P < 0.000 1) and upregulated the expression of apoptosis-related proteins cleaved caspase 3 (F=46.98, P=0.001 4) and caspase 3 (F=33.35, P=0.002 7).@*Conclusion@#Overexpression of TPX2 was found in TSCC tissues. Silencing of TPX2 might inhibit cell proliferation and promote cell apoptosis. TPX2 could be a new target for gene therapy of TSCC.