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1.
J Photochem Photobiol B ; 202: 111720, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31841988

RESUMO

It has been widely reported that ultraviolet-B (UV-B) radiation is the main extrinsic etiological agent that causes skin photodamage. UV-B exposure mediated photodamage (photo-aging/photo-carcinogenesis) to human skin is caused due to several physiological events at tissue, cellular and molecular levels that lead to impairment of skin function and integrity. In the present study, we investigated the protective role of Trigonelline (TG) against UV-B induced photo-damage in Human Dermal Fibroblasts (Hs68 cells) and Balb/C mice. We exposed human skin fibroblasts and Balb/C mice to UV-B radiation and evaluated various parameters of cellular damage, including, oxidative stress, cytosolic calcium (Ca2+) levels, apoptotic and ER-stress marker proteins. We found that UV-B irradiation induced ROS generation lead to the depletion of endoplasmic reticulum (ER) calcium and increased the expression of ER stress protein markers (phosphorylated elf2α, CHOP, ATF4) as well as apoptotic protein markers (Bcl2, Bax and caspase-9) in a dose and time dependent manner in Hs68 cells. We then determined the effect of TG treatment on UV-B -induced cell death in Hs68 cells and observed that cells exposed to UV-B radiation and treated with TG had a significantly higher survival rate compared to cells exposed to UV-B radiation alone. TG treatment successfully reduced oxidative stress; restored Ca2+ homeostasis and re-established the ER function and prevented apoptotic cell death process. Our results suggest that TG can be used as a potential therapeutic/cosmeceutic agent in preventing skin photo-damage.


Assuntos
Alcaloides/farmacologia , Apoptose/efeitos dos fármacos , Cálcio/metabolismo , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Substâncias Protetoras/farmacologia , Raios Ultravioleta , Animais , Apoptose/efeitos da radiação , Caspase 9/genética , Caspase 9/metabolismo , Linhagem Celular , Retículo Endoplasmático/metabolismo , Estresse do Retículo Endoplasmático/efeitos da radiação , Fator de Iniciação 1 em Eucariotos/genética , Fator de Iniciação 1 em Eucariotos/metabolismo , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Fibroblastos/efeitos da radiação , Expressão Gênica/efeitos dos fármacos , Expressão Gênica/efeitos da radiação , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Estresse Oxidativo/efeitos da radiação , Espécies Reativas de Oxigênio/metabolismo , Fator de Transcrição CHOP/genética , Fator de Transcrição CHOP/metabolismo
2.
J Photochem Photobiol B ; 203: 111749, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31884347

RESUMO

Gastric cancer (GC) is mainly widespread gastrointestinal malignancy,which reports for 8% of overallcases in carcinogenesis and 10% of yearly fatality, is 4thprimary cause of cancer associated death global. The plan of the present research was to develop ethanolic extract of Vitex negundo-loaded gold nanoparticles (VN-AuNPs) and to appraise the various characteristic methods likes UV-vis spectroscopy, SAED, FTIR, XRD and HR-TEM. Additionally, the anticancer effect of VN-AuNPs on AGS cells were analysed by cell viability, apoptotic morphological changes by TUNEL, AO/EtBr and Hoechst staining, alterations of mitochondrial membrane potential (MMP) and production reactive oxygen species (ROS). Moreover, the status of apoptosis gene such as caspase-3, Bcl-2, Bcl-XL, Bax and caspase-9 expressions was analysed by using western and RT-PCR techniques. Synthesized AuNPs established by UV absorption peak of the highest at 538 and crystal nature of AuNPs was additionallyverifiedwith SAED and XRD. TEM images were illustrates size and morphological division of NPs. FTIR examinationscompletedalkene, carbodiimide and aliphatic primary amines of biomolecules werepresent in synthesized VN-AuNPs. Additionally, AuNPs were stimulatedapoptosis throughthe cytotoxicity effect,changes of MMP, generation of ROS, nuclear and apoptotic morphological alterationsvia TUNEL, AO/EtBr and Hoechst assay. Furthermore, molecular mechanisms also provoked apoptosis through modulating pro (caspase-3, Bax, Bid, caspase-9) and anti-apoptotic (Bcl-2 and Bcl-XL) mediators by western blotting and gene expression in AGS cells. This production of AuNPs from VN was eco-friendly, large-scaled up and easy.


Assuntos
Apoptose/efeitos dos fármacos , Ouro/química , Nanopartículas Metálicas/toxicidade , Vitex/química , Caspase 3/genética , Caspase 3/metabolismo , Caspase 9/genética , Caspase 9/metabolismo , Linhagem Celular Tumoral , Química Verde , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Nanopartículas Metálicas/química , Extratos Vegetais/química , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patologia , Vitex/metabolismo
3.
Chemotherapy ; 64(3): 119-128, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31661694

RESUMO

OBJECTIVE: To investigate whether TRIAP1inhibition affects the ovarian cancer cell resistance to cisplatin (DDP) via the Cyt c/Apaf-1/caspase-9 pathway by in vitro and in vivo experiments. METHODS: CCK8 assay was performed to find out how treatment with both TRIAP1 siRNA and DDP affects the cell viability of SKOV3 cells and DDP-resistant human ovarian carcinoma cell line SKOV3/DDP. SKOV3/DDP cells were transfected with control siRNA or TRIAP1 siRNA before 24 h of treatment with DDP (5 µg/mL). Flow cytometry was employed to detect cell apoptosis and Western blot to examine the expressions of Cyt c/Apaf-1/caspase-9 pathway-related proteins. SKOV3/DDP cells transfected with control siRNA or TRIAP1 siRNA were subcutaneously injected into BALB/c-nu/nu nude mice followed by the intraperitoneal injection of DDP (4 mg/kg). Cyt c/Apaf-1/caspase-9 pathway in transplanted tumors was detected by immunohistochemistry. RESULTS: TRIAP1 expression declined in SKOV3 cells when compared with SKOV3/DDP cells. The proliferation rate was lower in SKOV3/DDP cells transfected with TRIAP1 siRNA combined with treatment of DDP (1, 2, 4, 6, 8, 16, 32 µg/mL) than in those transfected with control siRNA. Moreover, the TRIAP1 siRNA group had an increased SKOV3/DDP cell apoptosis rate with the activation of the Cyt c/Apaf-1/caspase-9 pathway. During DDP treatment, nude mice in TRIAP1 siRNA group had slower growth and smaller size of transplanted tumor than those in control siRNA group, with increased expression of Cyt c, Apaf-1, and caspase-9. CONCLUSION: TRIAP1 inhibition may enhance the sensitivity of SKOV3/DDP cells to cisplatin via activation of the Cyt c/Apaf-1/caspase-9 pathway.


Assuntos
Cisplatino/farmacologia , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , RNA Interferente Pequeno/metabolismo , Transdução de Sinais/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Fator Apoptótico 1 Ativador de Proteases/genética , Fator Apoptótico 1 Ativador de Proteases/metabolismo , Caspase 9/genética , Caspase 9/metabolismo , Linhagem Celular Tumoral , Cisplatino/uso terapêutico , Citocromos c/genética , Citocromos c/metabolismo , Regulação para Baixo/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Feminino , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/antagonistas & inibidores , Peptídeos e Proteínas de Sinalização Intracelular/genética , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Neoplasias Ovarianas/tratamento farmacológico , Neoplasias Ovarianas/metabolismo , Neoplasias Ovarianas/patologia , Interferência de RNA , RNA Interferente Pequeno/uso terapêutico , Transplante Heterólogo
4.
Molecules ; 24(17)2019 Aug 29.
Artigo em Inglês | MEDLINE | ID: mdl-31470638

RESUMO

Despite significant advances in the understanding, prevention, and treatment of cancer, the disease continues to affect millions of people worldwide. Chemoradiation therapy is a rational approach that has already proven beneficial for several malignancies. However, the existence of toxicity to normal tissue is a serious limitation of this treatment modality. The aim of the present study is to investigate the ability of polar steroids from starfish Patiria (=Asterina) pectinifera to enhance the efficacy of radiation therapy in colorectal carcinoma cells. The cytotoxic activity of polar steroids and X-ray radiation against DLD-1, HCT 116, and HT-29 cells was determined by an MTS assay. The effect of compounds, X-ray, and their combination on colony formation was studied using the soft agar method. The molecular mechanism of the radiosensitizing activity of asterosaponin P1 was elucidated by western blotting and the DNA comet assay. Polar steroids inhibited colony formation in the tested cells, and to a greater extent in HT-29 cells. Asterosaponin P1 enhanced the efficacy of radiation and, as a result, reduced the number and size of the colonies of colorectal cancer cells. The radiosensitizing activity of asterosaponin P1 was realized by apoptosis induction through the regulation of anti- and pro-apoptotic protein expression followed by caspase activation and DNA degradation.


Assuntos
Antineoplásicos/farmacologia , Apoptose/genética , Asterina/química , Regulação Neoplásica da Expressão Gênica , Compostos Policíclicos/farmacologia , Radiossensibilizantes/farmacologia , Saponinas/farmacologia , Animais , Antineoplásicos/química , Antineoplásicos/isolamento & purificação , Apoptose/efeitos dos fármacos , Apoptose/efeitos da radiação , Caspase 3/genética , Caspase 3/metabolismo , Caspase 9/genética , Caspase 9/metabolismo , Linhagem Celular Tumoral , Terapia Combinada , Células HCT116 , Células HT29 , Humanos , Compostos Policíclicos/química , Compostos Policíclicos/isolamento & purificação , Radiossensibilizantes/química , Radiossensibilizantes/isolamento & purificação , Saponinas/química , Saponinas/isolamento & purificação , Ensaio Tumoral de Célula-Tronco , Raios X , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo , Proteína bcl-X/genética , Proteína bcl-X/metabolismo
5.
Artif Cells Nanomed Biotechnol ; 47(1): 2737-2745, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31304798

RESUMO

Hepatocellular carcinoma is the most common liver cancer among different types of cancers. Cordyceps Militaris mushroom species traditionally used as an alternative medicine in china for centuries. Gold nanoparticles plays vital role in the development of the anticancer drugs. In our research, we investigated the gold nanoparticles with C. Militaris on the hepatocellular carcinoma HepG2 cells. The synthesized gold nanoparticles stability and integrity was studied at different time intervals. The gold nanoparticles potentially halt the growth of the HepG2 cells at the IC50 concentration between 10 µg and 12.5 µg/ml. The HR-TEM and XRD revealed the size and shape of the synthesized gold nanoparticles. The size of the gold nanoparticles was about 15 20 nm and the shape of gold nanoparticles was face-center-cubic structure. The FT-IR results proved that the gold nanoparticles contain hydroxyl and alkynes groups. The gold nanoparticles extract develops ROS and cause damage to the mitochondrial membrane potential in the hepatocellular carcinoma HepG2 cells. The gold nanoparticles extract tends to initiate the apoptosis by activating the Bax, Bid, caspases and inhibits the activation anti-apoptotic bcl-2 in the HepG2 cells. Our results concluded that the gold nanoparticles with C. Militaris would be an efficient chemotherapeutic drug against the hepatocellular carcinoma cells.


Assuntos
Cordyceps/química , Ouro/química , Ouro/farmacologia , Neoplasias Hepáticas/patologia , Nanopartículas Metálicas , Extratos Vegetais/química , Antineoplásicos/síntese química , Antineoplásicos/química , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Caspase 3/genética , Caspase 9/genética , Técnicas de Química Sintética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Química Verde , Células Hep G2 , Humanos , Concentração Inibidora 50 , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Nanotecnologia , Proteínas Proto-Oncogênicas c-bcl-2/genética , Espécies Reativas de Oxigênio/metabolismo , Proteína X Associada a bcl-2/genética
6.
Med Sci (Paris) ; 35(6-7): 497-500, 2019.
Artigo em Francês | MEDLINE | ID: mdl-31274074

Assuntos
Anticorpos Monoclonais/uso terapêutico , Neoplasias Hematológicas/terapia , Imunoterapia Adotiva/métodos , Proteína Acessória do Receptor de Interleucina-1/metabolismo , Linfócitos do Interstício Tumoral/metabolismo , Receptores de Antígenos Quiméricos/metabolismo , Animais , Anticorpos Monoclonais/genética , Antígenos de Neoplasias/imunologia , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/imunologia , Biomarcadores Tumorais/metabolismo , Caspase 9/genética , Proteínas de Fusão bcr-abl/metabolismo , Regulação Neoplásica da Expressão Gênica , Genes Transgênicos Suicidas , Células-Tronco Hematopoéticas/citologia , Células-Tronco Hematopoéticas/metabolismo , Humanos , Imunoterapia Adotiva/efeitos adversos , Proteína Acessória do Receptor de Interleucina-1/genética , Proteína Acessória do Receptor de Interleucina-1/imunologia , Células Matadoras Naturais/imunologia , Leucemia Mielogênica Crônica BCR-ABL Positiva/genética , Leucemia Mielogênica Crônica BCR-ABL Positiva/metabolismo , Leucemia Mielogênica Crônica BCR-ABL Positiva/patologia , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/metabolismo , Leucemia Mieloide Aguda/terapia , Linfócitos do Interstício Tumoral/imunologia , Linfócitos do Interstício Tumoral/transplante , Camundongos , Camundongos Endogâmicos BALB C , Neoplasias/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras/terapia , Receptores de Antígenos Quiméricos/imunologia
7.
Food Funct ; 10(7): 3868-3879, 2019 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-31184641

RESUMO

Lycopene (LYC) has been reported to exhibit antioxidant and immunoprotective activities, and our previous studies confirmed that LYC can alleviate multiple tissue damage induced by aflatoxin B1 (AFB1). However, it is unclear whether LYC could relieve the AFB1-induced immunosuppression. Thus, forty-eight male mice were randomly allocated and treated with LYC (5 mg kg-1) and/or AFB1 (0.75 mg kg-1) by intragastric administration for 30 days. We found that LYC alleviated AFB1-induced immunosuppression by relieving splenic structure injury and increasing the spleen weight, spleen coefficient, T lymphocyte subsets, the contents of IL-2, IFN-γ and TNF-α in serum, as well as the mRNA expression of IL-2, IFN-γ and TNF-α in spleen. Furthermore, LYC inhibited oxidative stress induced by AFB1via decreasing the levels of reactive oxygen species (ROS), hydrogen peroxide (H2O2) and malondialdehyde (MDA), while enhancing the total antioxidant capacity (T-AOC) and antioxidant enzyme activities. In addition, LYC also restrained splenic apoptosis through blocking mitochondria-mediated apoptosis in AFB1 intoxicated mice, presenting as the increase of mitochondrial membrane potential, and the decrease of cytoplasmic Cyt-c protein expression, cleaved Caspase-3 protein expression, Caspase-3/9 activities and mRNA expressions, as well as balancing the mitochondrial protein and mRNA expressions of Bax and Bcl-2. These results indicate that LYC can alleviate AFB1-induced immunosuppression by inhibiting oxidative stress and mitochondria-mediated apoptosis of mice spleen.


Assuntos
Aflatoxina B1/efeitos adversos , Apoptose/efeitos dos fármacos , Imunossupressão , Licopeno/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Baço/efeitos dos fármacos , Baço/metabolismo , Actinas/genética , Actinas/metabolismo , Animais , Caspase 3/genética , Caspase 3/metabolismo , Caspase 9/genética , Caspase 9/metabolismo , Modelos Animais de Doenças , Expressão Gênica/efeitos dos fármacos , Peróxido de Hidrogênio , Interferon gama/sangue , Interferon gama/genética , Interferon gama/metabolismo , Interleucina-2/sangue , Interleucina-2/genética , Interleucina-2/metabolismo , Masculino , Malondialdeído/metabolismo , Camundongos , Mitocôndrias/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , RNA Mensageiro/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Baço/lesões , Baço/patologia , Linfócitos T , Fator de Necrose Tumoral alfa/sangue , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo
8.
Medicina (Kaunas) ; 55(6)2019 May 31.
Artigo em Inglês | MEDLINE | ID: mdl-31159239

RESUMO

Background and objectives: Inflammatory bowel disease (IBD) mainly includes Crohn's disease (CD) and ulcerative colitis (UC). Both conditions are associated with an exacerbated intestinal immune response to harmless stimuli, leading to upregulation of pro-inflammatory mediators. Materials and Methods: The subjects of the study were 55 patients with IBD. The control group consisted of 35 healthy subjects. The researched material consisted of peripheral blood lymphocytes collected from the subjects. Expression of the genes BAX, BCL2, CASP3 and CASP9 was assessed at the mRNA level in the peripheral blood lymphocytes of patients with ulcerative colitis and Crohn's disease relative to the healthy subjects. The expression of the genes was determined by rtPCR using TaqMan probes specific for these genes. Results: The group of patients diagnosed with CD had statistically significantly higher expression of the genes BAX (p = 0.012), BCL2 (p = 0.022), CASP3 (p = 0.003) and CASP9 (p = 0.029) than healthy subjects. Expression of BAX, BCL2, CASP3 and CASP9 in UC patients in the active phase of the disease was significantly lower than in patients in remission: BAX (p = 0.001), BCL2 (p = 0.038) and CASP9 (p = 0.007). In patients with UC, the BAX/BCL2 ratio was significantly correlated (r = 0.473) with the duration of the disease. In the group of CD patients treated biologically, a significantly lower BAX/BCL2 ratio was demonstrated than in patients that were not biologically treated. Conclusions: Our research has shown a simultaneous increase in the expression of the anti-apoptotic BCL2 gene and the proapoptotic BAX gene, which suggests the dysregulation of apoptosis mechanisms in IBD. Significantly higher expression of BAX and BCL2 in UC patients in remission as compared to CD may suggest differences in these diseases in terms of prognosis and treatment. Our results may suggest that an underlying imbalance in factors controlling apoptosis in peripheral blood lymphocytes may be the response of the immune system to inflammation of the intestinal mucosa. Modulation of apoptosis may become an important therapeutic mechanism in IBD.


Assuntos
Morte Celular/fisiologia , Colite Ulcerativa/sangue , Doença de Crohn/sangue , Linfócitos/patologia , Adulto , Idoso , Caspase 3/genética , Caspase 9/genética , Morte Celular/genética , Colite Ulcerativa/genética , Colite Ulcerativa/fisiopatologia , Doença de Crohn/genética , Doença de Crohn/fisiopatologia , Feminino , Humanos , Mucosa Intestinal/fisiopatologia , Masculino , Pessoa de Meia-Idade , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteína X Associada a bcl-2/genética
9.
Mol Med Rep ; 20(2): 1451-1458, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31173227

RESUMO

Preeclampsia (PE) is a complication of pregnancy, and a leading cause of maternal mortality and morbidity worldwide. Recently, the dysregulation of long non­coding RNAs (lncRNAs) has been reported to contribute to the pathogenesis and progression of PE. This study aimed to examine the alterations in the lncRNA family with sequence similarity 99 member A (FAM99A) in PE and its effects on trophoblasts. The results of reverse transcription­quantitative PCR indicated that the expression levels of FAM99A were downregulated in placental tissues from women with severe PE compared with in those from controls. A Transwell invasion assay and wound healing assay revealed that overexpression of FAM99A promoted invasion and migration of HTR­8/SVneo cells; conversely, knockdown of FAM99A suppressed the invasive and migratory abilities of HTR­8/SVneo cells. Flow cytometry demonstrated that FAM99A overexpression induced a decrease in the apoptotic rate of cells, whereas knockdown of FAM99A increased the apoptotic rate of HTR­8/SVneo cells. Western blot analysis revealed that overexpression of FAM99A decreased the protein expression levels of cleaved caspase­3, cleaved caspase­9 and Bax, and increased Bcl­2 protein expression, whereas knockdown of FAM99A had the opposite effects on these protein levels. Overexpression of FAM99A also decreased caspase­3 activity in HTR­8/SVneo cells; however, knockdown of FAM99A increased caspase­3 activity. In addition, overexpression of FAM99A enhanced Wnt/ß­catenin signaling activity, whereas FAM99A knockdown exerted an inhibitory effect on the Wnt/ß­catenin signaling activity in HTR­8/SVneo cells. In conclusion, these results indicated that FAM99A may serve a role in modulating the functions of trophoblasts, partially via targeting Wnt/ß­catenin signaling.


Assuntos
Pré-Eclâmpsia/genética , RNA Longo não Codificante/genética , Trofoblastos/metabolismo , Via de Sinalização Wnt/genética , Adulto , Apoptose/efeitos dos fármacos , Estudos de Casos e Controles , Caspase 3/genética , Caspase 3/metabolismo , Caspase 9/genética , Caspase 9/metabolismo , Linhagem Celular , Movimento Celular/efeitos dos fármacos , Feminino , Regulação da Expressão Gênica , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/farmacologia , Pré-Eclâmpsia/metabolismo , Pré-Eclâmpsia/patologia , Gravidez , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , RNA Longo não Codificante/antagonistas & inibidores , RNA Longo não Codificante/metabolismo , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Trofoblastos/efeitos dos fármacos , Trofoblastos/patologia , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo , beta Catenina/genética , beta Catenina/metabolismo
10.
Mol Med Rep ; 20(2): 895-902, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31173258

RESUMO

Previous studies demonstrated that uropathogenic Escherichia coli infection contributes to human bacterial prostatitis. Apoptosis of prostate epithelial cells is closely associated with the progression of bacterial prostatitis. The aim of the present study was to investigate the effect of cellular tumor antigen p53 (p53) on the apoptosis of bacterial prostatitis cells. The prostate epithelial RWPE­1 cell line was infected with Escherichia coli, and treated cells and the culture supernatant were obtained at specific time points. The cell apoptosis rates, protein and mRNA of p53 were detected in the different treatment groups. Flow cytometry and terminal deoxynucleotidyl­transferase­mediated dUTP nick end labeling assays were used for the detection of cell apoptosis, and cell proliferation was determined by a Cell Counting Kit­8 assay. The expression of p53 was inhibited by small interfering (si)RNA, and its mRNA and protein were detected. An ELISA was used for detecting cytokines in the culture supernatant. The result demonstrated that Escherichia coli infection led to an increase in prostate epithelial cell apoptosis (P<0.05), and resulted in increases of interleukin (IL)­4, IL­6 and IL­8, and decrease in IL­10. p53, apoptosis regulator BAX (Bax), caspase­9 and Caspase­3 expression were upregulated upon Escherichia coli exposure (P<0.05). Following transfection with p53 siRNA, the promotion of cell apoptosis induced by Escherichia coli infection was decreased, and the p53 and Bax protein expression were additionally decreased. Therefore, it was suggested that Escherichia coli increases cell apoptosis in bacterial prostatitis by activating the death receptor pathway involving p53. Inhibition of p53 alleviated prostate cell apoptosis induced by Escherichia coli.


Assuntos
Apoptose/genética , Células Epiteliais/metabolismo , Interações Hospedeiro-Patógeno/genética , Proteína Supressora de Tumor p53/genética , Escherichia coli Uropatogênica/patogenicidade , Caspase 3/genética , Caspase 3/metabolismo , Caspase 9/genética , Caspase 9/metabolismo , Linhagem Celular , Células Epiteliais/microbiologia , Células Epiteliais/patologia , Regulação da Expressão Gênica , Humanos , Interleucina-10/genética , Interleucina-10/metabolismo , Interleucina-4/genética , Interleucina-4/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Interleucina-8/genética , Interleucina-8/metabolismo , Masculino , Próstata/microbiologia , Próstata/patologia , RNA Mensageiro/antagonistas & inibidores , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Transdução de Sinais , Proteína Supressora de Tumor p53/antagonistas & inibidores , Proteína Supressora de Tumor p53/metabolismo , Escherichia coli Uropatogênica/crescimento & desenvolvimento , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo
11.
Mol Med Rep ; 20(1): 135-140, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31115517

RESUMO

Toosendanin, a triterpenoid extracted from the root bark of Melia toosendan, has its origin from traditional Chinese medicine and has been used as a non­polluting and pesticide­free plant insecticide in China for fruit and vegetable production. In recent years, toosendanin has been found to inhibit tumor cell proliferation and promote tumor cell apoptosis. Ewing's sarcoma (ES) is the second most common primary malignant bone and soft tissue tumor in children and adolescents. Although the overall prognosis of ES has improved, the 5­year survival rate has not significantly increased. To analyze the role of toosendanin on ES progression, CCK­8 viability assay, flow cytometry, Hoechst 33258 staining and western blotting were performed. The present results suggested that toosendanin suppressed cell viability and induced apoptosis in human SK­ES­1 cells compared with DMSO treatment. In addition, in the present study, toosendanin was found to upregulate the expression of Bax and downregulate the expression of Bcl­2, altering the Bax/Bcl­2 ratio. Additionally, toosendanin promoted the release of cytochrome c, resulting in the activation of the mitochondrial apoptotic pathway, thus inducing the activation of caspase­9 and caspase­3, and the cleavage of PARP. Our results demonstrated that toosendanin inhibited the growth of ES cells in a dose­dependent manner and triggered mitochondrial apoptotic pathway to induce apoptosis. Therefore, toosendanin can potentially be utilized as an anticancer botanical drug for the treatment of ES.


Assuntos
Proliferação de Células/efeitos dos fármacos , Medicamentos de Ervas Chinesas/farmacologia , Mitocôndrias/efeitos dos fármacos , Sarcoma de Ewing/tratamento farmacológico , Apoptose/efeitos dos fármacos , Caspase 3/genética , Caspase 9/genética , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Mitocôndrias/genética , Proteínas Proto-Oncogênicas c-bcl-2/genética , Sarcoma de Ewing/genética , Sarcoma de Ewing/patologia , Transdução de Sinais/efeitos dos fármacos , Proteína X Associada a bcl-2/genética
12.
Anticancer Res ; 39(5): 2437-2441, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31092436

RESUMO

BACKGROUND/AIM: Non-small cell lung cancer (NSCLC) is one of the most common forms of lung cancer and the leading cause of cancer-related deaths in the world. Caspase 9 (CASP9) plays a central role in the intrinsic apoptotic pathway. The aim of the study was to investigate the role of caspase 9 gene polymorphism in patients with non-small cell lung cancer. MATERIALS AND METHODS: The study included 96 NSCLC cases and 67 controls. CASP9 Ex5+32 G>A polymorphism was investigated by real-time polymerase chain reaction. RESULTS: There was a significant difference between the groups in the frequency of CASP9 genotypes (p=0.008). The number of the carriers of the ancestral GG genotype, was significantly higher in the NSCLC group than in the control (p=0.009). The heterozygote GA genotype and mutant A allele frequency were significantly higher in the control group compared to the NSCLC group (p=0.005, p=0.009, respectively). Serum CASP9 levels were significantly lower in the patients group than in the control group (p<0.0001). CONCLUSION: CASP9 Ex5+32 GG genotype was a risk factor whereas the variant A allele could be a risk-reducing factor for NSCLC.


Assuntos
Carcinoma Pulmonar de Células não Pequenas/genética , Caspase 9/genética , Estudos de Associação Genética , Predisposição Genética para Doença , Adulto , Idoso , Alelos , Carcinoma Pulmonar de Células não Pequenas/sangue , Carcinoma Pulmonar de Células não Pequenas/patologia , Caspase 9/sangue , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único/genética , Fatores de Risco
13.
Hum Exp Toxicol ; 38(9): 1069-1081, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31117840

RESUMO

Apoptosis induction in cancer cells is one of the most efficient ways to treat cancer and find anticancer compounds. The aim of this study was to evaluate the cytotoxic effects of heat-killed indigenous probiotic bacteria and apoptosis induction in the HT-29 human colon adenocarcinoma cell line. The growth-inhibitory effects of probiotic heat-killed Lactobacillus brevis and Lactobacillus paracasei isolated from the traditional Iranian food "Terxine" on the HT-29 cell line were determined by 3-(4,5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) assay. Flow cytometry by Annexin-FITC kit, DNA fragmentation assay, 4,6-diamidino-2-phenylindole staining, and the expression of Bax, Bcl2, caspase-3, and caspase-9 were used to analyze apoptosis. MTT results demonstrated that the heat-killed bacteria inhibited the proliferation of HT-29 cells and induced apoptosis in a time-, dose-, and strain-dependent manner. The results demonstrated that both bacteria could induce apoptosis in the HT-29 cell line. Heat-killed probiotic bacteria increased the expression of Bax, caspase-3, and caspase-9 mRNA levels in HT-29 cell lines. Also, heat-killed probiotic bacteria reduced the expression of Bcl2 in HT-29 cells. The heat-killed probiotic bacteria in this study exhibited potent growth inhibitory effects on cancer cells. The results demonstrated that L. brevis has a greater ability to inhibit the growth of HT-29 cells and induce apoptosis, compared with L. paracasei. It is proposed that these bacteria can be used as biological products for the treatment and prevention of cancer, pending further investigation.


Assuntos
Adenocarcinoma/terapia , Neoplasias do Colo/terapia , Lactobacillus brevis , Lactobacillus paracasei , Probióticos/uso terapêutico , Adenocarcinoma/genética , Apoptose , Caspase 3/genética , Caspase 9/genética , Neoplasias do Colo/genética , Células HT29 , Temperatura Alta , Humanos , Proteínas Proto-Oncogênicas c-bcl-2/genética , Transdução de Sinais
14.
Biomed Pharmacother ; 115: 108876, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31022596

RESUMO

BACKGROUND: Rheumatoid arthritis (RA) is a common inflammatory disease, which significantly reduces the quality of life and increases the risk of cardiovascular and cerebrovascular diseases. The present work studied the therapeutic potency of Salvianolic acid B (Sal-B) for RA and revealed one of the possible underlying mechanisms. METHODS: Human rheumatoid fibroblast-like synoviocytes (MH7 A) were treated with Sal-B before, during or after lipopolysaccharide (LPS) stimulation. CCK-8 assay, Annexin V-FITC/PI double-staining, RT-qPCR, Western blotting and ELISA were carried out to measure the changes of cell viability, apoptosis, and the release of pro-inflammatory cytokines. Next, the involvement of miR-142-3p and related signaling pathways in Sal-B-mediated protection was studied. RESULTS: Sal-B (10 µM) treatment significantly ameliorated LPS injury to MH7 A cells, as cell viability was increased, expression of p53 and p21 was repressed, apoptosis was inhibited, and the release of MCP-1, IL-6 and TNF-α was reduced. However, Sal-B (10 µM) treated alone has no impacts on MH7 A cells in the abovementioned aspects. miR-142-3p was down-regulated by LPS stimulation, while was up-regulated by treatment of Sal-B. Rescue assay results showed that Sal-B did not remit LPS injury when miR-142-3p was silenced. And also, the inhibitory effects of Sal-B on NF-κB and JNK pathways were abolished by miR-142-3p silence. CONCLUSION: Sal-B could protect against and reverse LPS-induced injury in MH7 A cells, showing anti-apoptotic and anti-inflammatory capacities. The anti-RA potential of Sal-B might be via up-regulating miR-142-3p, and subsequently modulating NF-κB and JNK pathways.


Assuntos
Benzofuranos/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Lipopolissacarídeos/toxicidade , MicroRNAs/metabolismo , Sinoviócitos/efeitos dos fármacos , Benzofuranos/química , Caspase 3/genética , Caspase 3/metabolismo , Caspase 9/genética , Caspase 9/metabolismo , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Inibidor de Quinase Dependente de Ciclina p21/genética , Inibidor de Quinase Dependente de Ciclina p21/metabolismo , Citocinas/genética , Citocinas/metabolismo , Citoproteção/efeitos dos fármacos , Humanos , MicroRNAs/genética , Estrutura Molecular , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Sinoviócitos/metabolismo , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , Regulação para Cima
15.
Nanomedicine (Lond) ; 14(8): 1033-1047, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30925115

RESUMO

AIM: To induce a safe death to MCF-7 human breast cancer cell line through gene therapy based on iC9 suicide gene. MATERIALS & METHODS: To induce apoptosis to MCF-7 cell line, iC9 gene was transfected using pyridine-functionalized multi-walled carbon nanotubes. Then, to enhance chemotherapy, iC9 suicide gene therapy was performed alongside. RESULTS: The results show that the MCF-7 cells were efficiently eliminated in a high percentage by this approach. Furthermore, the suicide gene by itself/in combination with the chemotherapeutic drugs managed to pass the cell cycle arrests. CONCLUSION: We introduced an in vitro treatment approach based on suicide gene therapy and the first step was taken toward the enhancement of chemotherapy, although more investigation is required.


Assuntos
Neoplasias da Mama/terapia , Caspase 9/genética , Terapia Genética/métodos , Vetores Genéticos/química , Nanotubos de Carbono/química , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Sequência de Bases , Caspase 9/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Terapia Combinada/métodos , Dactinomicina/farmacologia , Doxorrubicina/farmacologia , Escherichia coli , Feminino , Humanos , Células MCF-7 , Piridinas/química , Transfecção/métodos
16.
Virus Res ; 265: 104-114, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30910697

RESUMO

Enterovirus 71 (EV71) causes hand-foot-and-mouth disease and severe neural complications in infants and young children. Viral pathogenesis is associated with virus-induced cell death and inflammatory cytokine production, which is usually correlated with the type of programmed cell death. EV71-infected cells were analyzed through microscopy, cell staining, and immunoblotting to determine the characteristics of EV71-induced cell death. Results demonstrated that EV71 infection induced cell shrinkage, nuclear condensation, decreased mitochondrial potential, and membrane phosphatidylserine translocation. Caspase-9 activation, poly (ADP-ribose) polymerase cleavage, and lactate dehydrogenase release were also observed during virus-induced cell death. The activated gasdermin D (GSDMD) and the phosphorylated mixed lineage kinase domain-like protein (p-MLKL) were not detected. These observations indicated that EV71-induced cell death was mainly executed by apoptosis through the intrinsic pathway rather than by GSDMD-mediated pyroptosis and p-MLKL-mediated necroptosis. Genome scanning analysis identified that EV71 2A, 2B, and 3C might be the determinant genes of virus-induced cell death. Further experiments showed that EV71 2A- and 3C-induced cell death exhibited dependence on their protease activities but involved different mechanisms. EV71 2A-induced cell death was correlated with the shut-off of host cap-dependent translation, whereas EV71 3C-induced cell death might not be ascribed to this mechanism. These findings would enhance our understanding of EV71 infection and viral pathogenesis, and help identify antiviral targets.


Assuntos
Apoptose/genética , Morte Celular/genética , Enterovirus Humano A/genética , Enterovirus Humano A/patogenicidade , Genes Virais , Caspase 9/genética , Linhagem Celular , Interações Hospedeiro-Patógeno , Humanos , Poli(ADP-Ribose) Polimerase-1/genética , Proteínas Virais/genética
17.
Fish Shellfish Immunol ; 88: 496-507, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30826414

RESUMO

In this study, two experiments were performed to explore the effect of Radix Bupleuri extracts (RBE) on growth, lipid deposition and metabolism and immune response of hybrid grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀) using in vitro and in vivo models. In vitro, we used 2 ml/L 20% lipid emulsion (LE)-induced steatosis in hybrid grouper primary hepatocytes, then RBE (200, 400 and 800 µg/ml) was added to the hepatocytes after (post-treatment) the incubation with 20% LE (2 ml/L) in the culture medium. We found that RBE markedly increased cell viability, which were consistent with hepatocytes morphological structure examination and lipid metabolism and immune related genes study. The above result suggested that RBE has a protective effect on this model of hepatocytes damage. In vivo, five graded levels of RBE at 0, 200, 400, 800 and 1600 mg/kg diet were supplemented to a basal diet with 15% lipid levels (high lipid), and fed to a total of 300 hybrid grouper with an average initial weight of 25.58 ±â€¯0.05 g for 8 weeks. Growth performance, liver histology, plasma biochemical parameters, and expression of genes involved in lipid metabolism and immune-related were measured. The study indicated that dietary RBE significantly improved growth performance and feed utilization and reduced hepatosomatic index. Dietary supplementation with 200-800 mg/kg RBE diets effectively decreased serum ALP, ALT, AST and LDH contents in fish. Furthermore, adipogenesis relative mRNA levels of DGAT2, G6PD, ME1 and DGKα in fish fed 200-400 mg/kg RBE diets were lower (P < 0.05) than in those fed RBE0 diets, while dietary supplementation with 200-800 mg/kg RBE diets up-regulated lipolysis-related genes (CPT1, LPL and PPARα) expression in the liver of hybrid grouper. Moreover, dietary RBE down-regulated the expression of apoptosis-related genes (caspase-9), up-regulated the expression of antioxidant genes (CAT) and immune-related genes (MHC2, IKKα and TGF-ß1). Thus, our data suggest that RBE suppressed lipid accumulation and enhanced immune capability in hybrid grouper both in vitro and in vivo. These results offer new insight into RBE as a hepatoprotective in fish.


Assuntos
Bass/imunologia , Suplementos Nutricionais/análise , Fígado Gorduroso/veterinária , Hepatócitos/metabolismo , Metabolismo dos Lipídeos , Extratos Vegetais/administração & dosagem , Ração Animal/análise , Animais , Apoptose , Bass/genética , Bass/crescimento & desenvolvimento , Caspase 9/genética , Sobrevivência Celular , Células Cultivadas , Fígado Gorduroso/tratamento farmacológico , Feminino , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Hibridização Genética , Lipólise , Fígado/imunologia , Fígado/metabolismo , Masculino , Raízes de Plantas/química , RNA Mensageiro
18.
Acta Otolaryngol ; 139(4): 336-339, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30907242

RESUMO

BACKGROUND: Ménière's disease (MD) is a disorder of the inner ear, causing episodes of vertigo. Although surgery is reserved for intractable MD, intratympanic gentamicin (ITG) injection has become an alternative for controlling vertigo. AIMS/OBJECTIVES: To investigate the genetic basis of ITG efficacy. MATERIAL AND METHODS: We hypothesized that single nucleotide polymorphisms (SNPs) affect outcomes in patients with MD who receive ITG injections. Whole-exome sequencing was used to determine variations in coding regions. RESULTS: Multivariate analysis revealed two SNPs, rs1052571 in caspase 9 (CASP9; p = .017) and rs3745274 in cytochrome P450 2B6 (p = .053), which were associated with susceptibility to ITG injections. Only the C-allele in the rs1052571 SNP was significantly associated with susceptibility (p = .027; odds ratio: 5.95; 95% confidence interval: 1.26-28.57, by Fisher's exact test). CONCLUSIONS AND SIGNIFICANCE: Our results elucidated the role of the rs1052571 SNP and provide a genetic perspective on gentamicin efficacy (susceptibility) in treating intractable MD.


Assuntos
Caspase 9/genética , Gentamicinas/uso terapêutico , Doença de Meniere/complicações , Inibidores da Síntese de Proteínas/uso terapêutico , Vertigem/tratamento farmacológico , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Seguimentos , Humanos , Injeção Intratimpânica , Masculino , Pessoa de Meia-Idade , Variantes Farmacogenômicos , Polimorfismo de Nucleotídeo Único , Vertigem/etiologia
19.
Mol Cell ; 74(1): 19-31.e7, 2019 04 04.
Artigo em Inglês | MEDLINE | ID: mdl-30878284

RESUMO

Viral infection triggers host defenses through pattern-recognition receptor-mediated cytokine production, inflammasome activation, and apoptosis of the infected cells. Inflammasome-activated caspases are known to cleave cyclic GMP-AMP synthase (cGAS). Here, we found that apoptotic caspases are critically involved in regulating both DNA and RNA virus-triggered host defenses, in which activated caspase-3 cleaved cGAS, MAVS, and IRF3 to prevent cytokine overproduction. Caspase-3 was exclusively required in human cells, whereas caspase-7 was involved only in murine cells to inactivate cGAS, reflecting distinct regulatory mechanisms in different species. Caspase-mediated cGAS cleavage was enhanced in the presence of dsDNA. Alternative MAVS cleavage sites were used to ensure the inactivation of this critical protein. Elevated type I IFNs were detected in caspase-3-deficient cells without any infection. Casp3-/- mice consistently showed increased resistance to viral infection and experimental autoimmune encephalomyelitis. Our results demonstrate that apoptotic caspases control innate immunity and maintain immune homeostasis against viral infection.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Apoptose , Caspases/metabolismo , Fator Regulador 3 de Interferon/metabolismo , Interferon Tipo I/metabolismo , Nucleotidiltransferases/metabolismo , Viroses/enzimologia , Proteínas Adaptadoras de Transdução de Sinal/genética , Animais , Caspase 2/genética , Caspase 2/metabolismo , Caspase 3/genética , Caspase 3/metabolismo , Caspase 7/genética , Caspase 7/metabolismo , Caspase 9/genética , Caspase 9/metabolismo , Caspases/genética , Feminino , Células HEK293 , Células HeLa , Interações Hospedeiro-Patógeno , Humanos , Imunidade Inata , Fator Regulador 3 de Interferon/genética , Masculino , Camundongos Endogâmicos C57BL , Nucleotidiltransferases/genética , Vírus Sendai/imunologia , Vírus Sendai/patogenicidade , Transdução de Sinais , Células THP-1 , Vírus Vaccinia/imunologia , Vírus Vaccinia/patogenicidade , Viroses/genética , Viroses/imunologia , Viroses/virologia
20.
Dis Markers ; 2019: 7378967, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30886656

RESUMO

Triple-negative breast cancer (TNBC) harbors genetic heterogeneity and generally has more aggressive clinical outcomes. As such, there is urgency in identifying new prognostic targets and developing novel therapeutic strategies. In this study, miR-224 was overexpressed in breast cancer cell lines and TNBC primary cancer samples. Knockdown of miR-224 in MDA-MB-231 cancer cells reduced cell proliferation, migration, and invasion. Through integrating in silico prediction algorithms with KEGG pathway and Gene Ontology analyses, CASP9 was identified to be a potential target of miR-224. miR-224 knockdown significantly increased CASP9 transcript and protein levels. Furthermore, luciferase reporter assays confirmed a direct interaction of miR-224 with CASP9. Our findings have demonstrated that the miR-224/CASP9 axis plays an important role in TNBC progression, providing evidence in support of a promising therapeutic strategy for this disease.


Assuntos
Carcinogênese/genética , Caspase 9/genética , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Neoplasias de Mama Triplo Negativas/genética , Caspase 9/metabolismo , Linhagem Celular , Movimento Celular , Proliferação de Células , Células Cultivadas , Regulação para Baixo , Feminino , Humanos , Células MCF-7 , MicroRNAs/metabolismo , Neoplasias de Mama Triplo Negativas/metabolismo , Neoplasias de Mama Triplo Negativas/patologia
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