RESUMO
Copper transporter 1 (CTR1) plays an important role in increasing cisplatin intake. Our previous studies showed that CTR1 expression was upregulated by (-)-epigallocatechin-3-gallate (EGCG), a green tea polyphenol, therefore enhanced cisplatin sensitivity in ovary cancer and non-small-cell lung cancer (NSCLC) cells. In the current study in the non-small-cell lung cancer cells, we uncovered a potential mechanism of EGCG-induced CTR1 through its pro-oxidative property. We found that EGCG increased reactive oxygen species (ROS) generation, while in the presence of ROS scavenger N-acetyl-cysteine (NAC), ROS production was eliminated. Changes of CTR1 expression were consistent with the ROS level. Simultaneously, EGCG downregulated ERK1/2 while upregulated lncRNA nuclear paraspeckle assembly transcript 1 (NEAT1) through ROS to induce CTR1 expression. Besides, in a nude mouse xenografts model, EGCG treatment raised ROS level, expression of CTR1 and NEAT1 in tumor tissue. Also, ERK1/2 and p-ERK1/2 were suppressed as well. Taken together, these results suggested a novel mechanism that EGCG mediated ROS to regulate CTR1 expression through the ERK1/2/NEAT1 signaling pathway, which provided more possibilities for EGCG as a natural agent in adjuvant therapy of lung cancer.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Catequina/análogos & derivados , Transportador de Cobre 1/genética , RNA Longo não Codificante/genética , Animais , Apoptose/efeitos dos fármacos , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Catequina/farmacologia , Linhagem Celular Tumoral , Cisplatino/farmacologia , Feminino , Humanos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Camundongos , Oxirredução/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Chá/química , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
This study aimed to measure the body composition, dietary patterns and its associated factors in medical students. A cross-sectional study was conducted among 695 students studying at Nanjing medical university, China. Data regarding dietary intake factors was collected by a validated food frequency questionnaire. Principle component analysis and multivariate linear regressions were used. Body composition including BMI and visceral fats index of the males were significantly higher (P < .05) than females. While body fats percentage and fats mass index/free fat mass index (FMI/FFMI) ratio of females was significantly higher (P < .05) than males. Three dietary patterns were identified: western dietary pattern, meat pattern, and vegetables and fruits pattern. The western pattern was having an independent negative association (P < .05) with age and financial status, while positive association (P < .05) with sleeping duration and FMI/FFMI ratio. Vegetables and fruits patterns was having positive association with physical exercise and while negative association with FMI/FFMI ratio. Meat pattern was having positive association with educational levels and sleeping duration, while negative association with physical exercise and FMI/FFMI ratio. In conclusion, medical students adopted less healthy dietary patterns as compared to healthy dietary patterns, which were found to be more associated with some adverse dietary and lifestyle behavior outcomes.
Assuntos
Composição Corporal/fisiologia , Dieta , Comportamento Alimentar/fisiologia , Estudantes de Medicina , Adolescente , Adulto , China , Coleta de Dados , Feminino , Humanos , Estilo de Vida , Masculino , Fatores Socioeconômicos , Inquéritos e Questionários , Adulto JovemRESUMO
Non-small-cell lung cancer (NSCLC) appears to be a significant threat to public health worldwide. MicroRNAs have been identified as significant regulators for the development of NSCLC. Previous reports have suggested that hsa-mir-485-5p is dysregulated in various cancers. RXRα, as a kind of nuclear receptor, is an effective target of cancer treatment. Cancer stem cells (CSCs) are recognized as the main cause for tumor metastasis, recurrence, and chemotherapy resistance. However, the mechanism by which hsa-mir-485-5p and RXRα modulate CSCs in NSCLC remains unknown. Here, we found that hsa-mir-485-5p was decreased in serum samples from patients with NSCLC and NSCLC cells. Meanwhile, epigallocatechin-3-gallate (EGCG), an effective anticancer compound extracted from green tea, can enhance hsa-mir-485-5p expression. Hsa-mir-485-5p mimics markedly inhibited NSCLC cell growth and induced cell apoptosis. However, inhibition of hsa-mir-485-5p significantly enriched CSC-like traits. Moreover, bioinformatics analysis predicted the binding correlation between hsa-mir-485-5p and RXRα, which was confirmed by a dual-luciferase reporter assay. We observed that RXRα was increased in NSCLC and EGCG could inhibit RXRα levels dose dependently. In addition, RXRα upregulation or activation expanded the CSC-like properties of NSCLC cells, whereas RXRα inhibition or inactivation could exert a reverse phenomenon. Consistently, in vivo experiments also validated that EGCG could repress the CSC-like characteristics by modulating the hsa-mir-485-5p/RXRα axis. Our findings may reveal a novel molecular mechanism for the treatment of NSCLC.
Assuntos
Anticarcinógenos/uso terapêutico , Carcinoma Pulmonar de Células não Pequenas/sangue , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Catequina/análogos & derivados , Neoplasias Pulmonares/sangue , Neoplasias Pulmonares/tratamento farmacológico , MicroRNAs/metabolismo , Células-Tronco Neoplásicas/metabolismo , Receptor X Retinoide alfa/metabolismo , Células A549 , Análise de Variância , Animais , Catequina/uso terapêutico , Regulação para Baixo , Técnicas de Silenciamento de Genes , Inativação Gênica , Células HEK293 , Humanos , Camundongos , Camundongos Nus , MicroRNAs/química , MicroRNAs/genética , Mimetismo Molecular/genética , Receptor X Retinoide alfa/genética , Transdução de Sinais/efeitos dos fármacos , Transfecção , Regulação para Cima , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Non-small cell lung cancer (NSCLC) remains one of the most aggressive tumors with low life expectancy worldwide. The existence of cancer stem cells (CSCs) contributes to the failure of cancer treatment resulted from drug resistance. Altered microRNA expression has been observed in human tumors due to its role in tumor growth, progression, and metastasis. Hence, the aim of our present study was to investigate the effects of miR-485 on the CSC-like traits in NSCLC A549-cisplatin resistant cells and concentrate on the underlying molecular mechanism. It was found that CSC-like phenotypes were much more enriched in A549/cisplatin (A549/CDDP) cells compared to A549-parental cells. In addition, we observed that miR-485 was greatly decreased in A549/CDDP cells and miR-485 overexpression was able to decrease the stemness of A549/DDP cells. Meanwhile, epigallocatechin-3-gallate (EGCG), a green tea polyphenol which has been identified as an effective anticancer compound was able to increase miR-485 expression dose-dependently in A549/CDDP cells. Inhibitors of miR-485 remarkably increased CSC-like phenotypes, which could be reversed by indicated doses of EGCG. Moreover, CD44 was predicted as downstream target of miR-485 and the correlation between them was validated by performing dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. Subsequently, in vivo experiments were employed to confirm that EGCG restrained CSC-like characteristics by increasing miR-485 and decreasing CD44 expression. Taken together, it was implied that stemness features and CSC population were suppressed by EGCG-modulated miR-485/CD44 axis in A549/CDDP cells.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Catequina/análogos & derivados , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Receptores de Hialuronatos/genética , Neoplasias Pulmonares/tratamento farmacológico , MicroRNAs/genética , Células-Tronco Neoplásicas/efeitos dos fármacos , Células A549 , Animais , Carcinoma Pulmonar de Células não Pequenas/genética , Catequina/administração & dosagem , Catequina/farmacologia , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cisplatino/administração & dosagem , Cisplatino/farmacologia , Relação Dose-Resposta a Droga , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Neoplasias Pulmonares/genética , Camundongos , Camundongos Nus , Células-Tronco Neoplásicas/metabolismo , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Apoptosis of vascular smooth muscle cells (VSMCs) accelerates manifestation of plaque vulnerability in atherosclerosis. Long noncoding RNA NEAT1 participates in the proliferation and apoptosis of cells. In addition, circadian clock genes play a significant role in cell apoptosis. However, whether acrolein, an environmental pollutant, affects the apoptosis of VSMCs by regulating NEAT1 and clock genes is still elusive. We established VSMCs as an atherosclerotic cell model in vitro. Acrolein exposure reduced survival rate of VSMCs, and raised apoptosis percentage through upregulating the expression of Bax, Cytochrome c and Cleaved caspase-3 and downregulating Bcl-2. Asparagus extract (AE), as a dietary supplementation, was able to protect VSMCs against acrolein-induced apoptosis. Expression of NEAT1, Bmal1 and Clock was decreased by acrolein, while was ameliorated by AE. Knockdown of NEAT1, Bmal1 or Clock promoted VSMCs apoptosis by regulating Bax, Bcl-2, Cytochrome c and Caspase-3 levels. Correspondingly, overexpression of NEAT1 inhibited the apoptosis. We also observed that silence of NEAT1 repressed the expression of Bmal1/Clock and vice versa. In this study, we demonstrated that VSMCs apoptosis induced by acrolein was associated with downregulation of NEAT1 and Bmal1/Clock. AE alleviated the effects of proapoptotic response and circadian disorders caused by acrolein, which shed a new light on cardiovascular protection.
Assuntos
Fatores de Transcrição ARNTL/genética , Acroleína/toxicidade , Apoptose , Proteínas CLOCK/genética , Miócitos de Músculo Liso/efeitos dos fármacos , Extratos Vegetais/farmacologia , RNA Longo não Codificante/genética , Asparagus/química , Técnicas de Silenciamento de Genes , HumanosRESUMO
OBJECTIVES: Time-restricted feeding (TRF) is a dietary therapeutic remedy for the prevention and treatment of metabolic diseases. Gut microbiota may influence the host metabolism and nutritional status of individuals. Given the significance of TRF and gut microbiota in metabolic diseases, the aim of this study was to explore the association between TRF and gut microbiota in healthy individuals, which is not clearly elucidated. METHODS: Thirty healthy men (18-30 y of age) were divided in to two groups (TRF: n = 15 and non-TRF: n = 15). The TRF group was instructed to not consume any food for 16 h/d. Two-day food diary was used for dietary data collection. Stool samples were collected from both groups after 25 d of TRF or non-TRF. Gut microbiota profile was analyzed and quantified by using 16S rRNA gene sequencing. RESULTS: Cluster analysis revealed that Prevotlla_9, Faecalibacterium, and Dialister were the most abundant species in TRF, whereas Prevotell_7, Alloprevotella, and Prevotella_2 were less abundant in the non-TRF group. At the genus level, gut microbiota of the TRF group was significantly changed compared with that of the non-TRF group. Moreover, bar plot analysis revealed that Bacteroidetes was the most abundant phylum in TRF group, followed by Firmicutes. Heat map correlation showed that polyunsaturated fatty acids and vitamin D were positively correlated with Firmicutes, whereas iodine, vitamin E, magnesium, and carbohydrate intake were negative correlated with microbial richness. CONCLUSION: The present study demonstrated that TRF is associated with microbial composition and relative abundance. TRF intervention might increase microbial abundance, thereby influencing the host metabolism and nutritional status.
Assuntos
Microbioma Gastrointestinal , Ingestão de Alimentos , Fezes , Firmicutes , Humanos , Masculino , RNA Ribossômico 16S/genéticaRESUMO
Circadian rhythm plays an important role in diverse physiological processes. Abnormal expression of circadian rhythm genes is associated with increased risk of disease, including different types of cancer. The cancer stem cell (CSC) hypothesis suggests that there is a small subset of stemlike cells within tumors that are responsible for tumor initiation. However, the biological effect of circadian rhythm on CSCs remains largely unknown. Studies have highlighted that the circadian rhythm protein CLOCK controls key aspects of various diseases. In the present study, lung cancer stemlike cells were successfully enriched using a sphere formation assay. Next, it was observed that CLOCK mRNA and protein expression levels in the A549 and H1299 sphere cells were notably increased compared with those in the corresponding parental cells. In addition, flow cytometry was performed to isolate CD133+ cells and, consistently, CLOCK expression was also found to be markedly upregulated in CD133+ lung cancer cells. Subsequently, to determine the effect of CLOCK on lung cancer stem cells in detail, CLOCK was knocked down using targeted short inhibiting RNA and the results demonstrated that the sphereforming ability of the A549 and H1299 cell lines was reduced. In addition, CSClike properties, including the expression of CD133, CD44, sex determining region Ybox 2, Nanog and octamerbinding transcription factor 4, were markedly decreased in the A549 and H1299 sphere cells following knockdown of CLOCK. Epigallocatechin3gallate (EGCG), a green tea polyphenol, has been reported to be a potential anticancer phytochemical. EGCG was found to repress CLOCK expression in A549 and H1299 sphere cells. In addition, EGCG also decreased the ratio of CD133+ cells. The Wnt/ßcatenin pathway was notably inactivated by the knockdown of CLOCK in A549 and H1299 sphere cells. Subsequently, using a xenograft model, it was demonstrated that EGCG suppressed the CSClike characteristics of lung cancer cells by targeting CLOCK. In conclusion, the present study demonstrated that EGCG inhibited the selfrenewal ability of lung cancer stemlike cells by targeting CLOCK.
Assuntos
Proteínas CLOCK/metabolismo , Catequina/análogos & derivados , Autorrenovação Celular/efeitos dos fármacos , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Células-Tronco Neoplásicas/metabolismo , Células-Tronco Neoplásicas/patologia , Animais , Catequina/farmacologia , Linhagem Celular Tumoral , Ritmo Circadiano/efeitos dos fármacos , Ritmo Circadiano/genética , Feminino , Humanos , Camundongos Endogâmicos BALB C , Camundongos Nus , Células-Tronco Neoplásicas/efeitos dos fármacos , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/genética , Via de Sinalização Wnt/efeitos dos fármacosRESUMO
Circadian rhythm is believed to play important roles in atherosclerosis. The gut microbiota is found to be closely related to atherogenesis, and shows compositional and functional circadian oscillation. However, it's still unclarified whether circadian clock and intestinal microbiota are involved in the progression of atherosclerosis induced by environmental pollutant acrolein. Herein, patients with atherosclerosis showed higher MMP9, a promising biomarker for atherosclerosis, and lower Bmal1 and Clock expression in the plasma. Interestingly, acrolein exposure contributed to the increased MMP9, decreased Clock and Bmal1, and activated MAPK pathways in human umbilical vein endothelial cells (HUVECs). We found that knockdown of Clock or Bmal1 lead to upregulation of MMP9 in HUVECs, and that Clock and Bmal1 expression was elevated while MAPK pathways were blocked. Atherosclerotic apolipoproteinE-deficient mice consumed a high-fat diet were used and treated with acrolein (3â¯mg/kg/day) in the drinking water for 12 weeks. Upregulation of MMP9, and downregulation of Clock and Bmal1 were also observed in plasma of the mice. Besides, acrolein feeding altered gut microbiota composition at a phylum level especially for an increased Firmicutes and a decreased Bacteroidetes. Additionally, gut microbiota showed correlation with atherosclerotic plaque, MMP9 and Bmal1 levels. Therefore, our findings indicated that acrolein increased the expression of MMP9 through MAPK regulating circadian clock, which was associated with gut microbiota regulation in atherosclerosis. Circadian rhythms and gut microbiota might be promising targets in the prevention of cardiovascular disease caused by environmental pollutants.
Assuntos
Fatores de Transcrição ARNTL/sangue , Aterosclerose/patologia , Proteínas CLOCK/sangue , Ritmo Circadiano/fisiologia , Microbioma Gastrointestinal/fisiologia , Metaloproteinase 9 da Matriz/metabolismo , Fatores de Transcrição ARNTL/genética , Acroleína , Adulto , Animais , Apolipoproteínas E/genética , Aterosclerose/induzido quimicamente , Proteínas CLOCK/genética , Linhagem Celular , Relógios Circadianos/fisiologia , Dieta Hiperlipídica , Regulação para Baixo , Feminino , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Humanos , Masculino , Camundongos , Camundongos KnockoutRESUMO
Long non-coding RNAs (lncRNAs) have been identified to exert crucial roles in tumorigenesis and can serve as novel biomarkers for cancer therapy including lung cancer. Cisplatin is a first-line chemotherapeutic agent in non-small cell lung cancer (NSCLC), but the therapeutic effect is unsatisfactory, partly due to drug resistance. Emerging evidence showed that chemo-resistance is associated with acquisition of cancer stem cell (CSC)-like properties. Cisplatin resistance remains a major obstacle in the treatment of lung cancer, and its mechanism is still not fully elucidated. Meanwhile, CSCs have been involved in tumor metastasis, tumor recurrence and chemotherapy resistance. So far, the mechanism of nuclear enriched abundant transcript 1 (NEAT1) in modulating CSCs in lung cancer remains barely known. Therefore, we aimed to explore the correlation between NEAT1 and cancer stem cells in lung cancer. In our current study, we observed that CSC-like traits were much more enriched in cisplatin-resistant A549/CDDP cells. In addition, NEAT1 was obviously up-regulated in A549/CDDP cells compared with parental A549â¯cells. Knockdown of NEAT1 decreased the CSC-like properties of A549/CDDP cells through inhibiting tumor cell sphere volume, repressing CSC-like biomarkers levels and restraining CD44 positive cell ratios. Oppositely, overexpression of NEAT1 enhanced the stemness respectively. Moreover, it has been reported that Wnt pathway is implicated in many vital cellular functions including cancer stem cells. Here, it was exhibited that Wnt signal pathway was inactivated by knockdown of NEAT1 whereas activated by NEAT1 overexpression in A549/CDDP cells. Taken these together, it was indicated that NEAT1 could exert a novel biological role in NSCLC chemo-resistance.