Objective: In this study, we aimed to examine the effect of MAN on human lung cancer and reveal the underlying molecular mechanism.
Methods: MTT assay was conducted to measure cell viability. Annexin V-FITC/PI staining was used to detect cell apoptosis. Confocal microscope was performed to determine the formation of autophagosomes and autolysosomes. Flow cytometry was performed to quantify cell death. Western blotting was used to determine the related-signaling pathway.
Results: In the present study, we demonstrated for the first time that MAN inhibitd cell proliferation and induced cell apoptosis in human non-small-cell lung carcinoma (NSCLC) cells. We found that MAN treatment dysregulated mitochondrial function and led to mitochondrial apoptosis in A549 and PC9 cells. Meanwhile, MAN enhanced autophagy flux by the increase of autophagosome formation, the fusion of autophagsomes and lysosomes and lysosomal function. Moreover, mTOR signaling pathway, a classical pathway regualting autophagy, was inhibited by MAN in a time- and dose-dependent mannner, resulting in autophagy induction. Interestingly, autophagy inhibition by CQ or Atg5 knockdown attenuated cell apoptosis by MAN, indicating that autophagy serves as cell death. Furthermore, autophagy-mediated cell death by MAN can be blocked by reactive oxygen species (ROS) scavenger NAC, indicating that ROS accumulation is the inducing factor of apoptosis and autophagy. In summary, we revealed the molecular mechanism of MAN against lung cancer through apoptosis and autophagy, suggesting that MAN might be a novel therapeutic agent for NSCLC treatment.
DISCUSSION: Within 2-3 days of treatment application, encouraging results were achieved in total dry solids (TDS), total suspended solid (TSS), turbidity, chemical oxygen demand (COD), specific resistance to filtration (SRF), and pH due to fungal treatment in recognition of bioseparation and dewaterability of wastewater sludge compared to control. The significant reduction of TDS was remarked at fungal biomass (FB) in wheat flour (WF) treatment. The removal of TSS, turbidity, COD, and SRF were observed 96.0%, 99.4%, 92.6%, and 97.6%, respectively, in supernatant at 5 days by FB in WF. The SRF measuring the dewaterability was decreased with maximum (0.26 × 10(-12) mg/kg) equivalent to 95.5% at 2 days in FB in WF also. FB in WF broth is a potential, environmental friendly, comparatively low-cost biological technique which might play the significant role for bioremediation and bioseparation of domestic wastewater sludge. The present technique may bring a dynamic change in treatment of wastewater in future.
DESIGN: Food choice was assessed using the validated New Zealand Adolescent FFQ. Principal components analysis was used to determine dietary patterns. Trained research assistants measured participants' height and body mass. Cardiorespiratory fitness was assessed in a subset of participants using the multistage 20 m shuttle run. The level and stage were recorded, and the corresponding VO2max was calculated. Differences in mean VO2max according to sex and BMI were assessed using t tests, while associations between cardiorespiratory fitness and dietary patterns were examined using linear regression analyses adjusted for age, sex, school attended, socio-economic deprivation and BMI.
SETTING: Secondary schools in Otago, New Zealand.
SUBJECTS: Students (n 279) aged 14-18 years who completed an online lifestyle survey during a class period.
RESULTS: Principal components analysis produced three dietary patterns: 'Treat Foods', 'Fruits and Vegetables' and 'Basic Foods'. The 279 participants who provided questionnaire data and completed cardiorespiratory fitness testing had a mean age of 15·7 (sd 0·9) years. Mean VO2max was 45·8 (sd 6·9) ml/kg per min. The 'Fruits and Vegetables' pattern was positively associated with VO2max in the total sample (β=0·04; 95%CI 0·02, 0·07), girls (β=0·06; 95% CI 0·03, 0·10) and boys (β=0·03; 95% CI 0·01, 0·05).
CONCLUSIONS: These results indicate that increase in cardiorespiratory fitness was associated with a healthier dietary pattern, suggesting both should be targeted as part of a global lifestyle approach. Longitudinal studies are needed to confirm this association in relation to health outcomes in New Zealand adolescents.
METHODS: SPF grade 4-6-week-old Kunming rats were randomly divided into 5 groups including a blank group, sham-operated group, model group, acupuncture, and moxibustion (AnM) group, and positive group. A total of 10 rats were included in each group. The model group, the AnM group, and the positive group were prepared by ligating the left sciatic nerve. AnM group was used for acupuncture and moxibustion therapy intervention, and the positive group was rendered to quick-acting sciatica pills once a day for 7 days (3 courses of treatment). The blank group, sham-operated group, and model group were not treated. The changes in thermal and mechanical pain thresholds were observed before and after the operation, and the morphological changes of the dorsal horn of the spinal cord in the lumbosacral region of the rats in each group were observed by HE staining after the courses of treatment finished. The contents of IL-1β, IL-6, IL-18, and TNF-α were measured by ELISA and the expressions of NOX1, NOX2, NOX4, and NLRP3 genes were detected by RT-qPCR while the protein expressions of NOX1, NOX2, NOX4 and NLRP3 were analyzed by Western blotting.
RESULTS: The AnM and positive group showed a significant increase in thermal and mechanical pain thresholds after treatment, while there was no significant change in the model group. As compared to the control group, the contents of IL- 1β, IL-6, IL-18, and TNF-α, as well as the relative expressions of NOX1, NOX2, NOX4, and NLRP3 genes were significantly increased in the model group (P
OBJECTIVE: In this study, we explained the development of graphene oxide/polyethylene glycol/folic acid/brucine nanocomposites (GO/PEG/Bru-FA NCs) and evaluated their antimicrobial and anticancer effect on the liver cancer HepG2 cells.
METHODOLOGY: The GO/PEG/Bru-FA NCs were prepared using the co-precipitation technique and characterized using various techniques. The cytotoxicity of the GO/PEG/Bru-FA NCs was tested against both liver cancer HepG2 and non-malignant Vero cells using an MTT assay. The antimicrobial activity of the GO/PEG/Bru-FA NCs was tested against several pathogens using the well diffusion technique. The effects of GO/PEG/Bru-FA NCs on endogenous ROS accumulation, apoptosis, and MMP levels were examined using corresponding fluorescent staining assays, respectively. The apoptotic protein expressions, such as Bax, Bcl-2, and caspases, were studied using the corresponding kits.
RESULTS: The findings of various characterization assays revealed the development of GO/PEG/Bru-FA NCs with face-centered spherical morphology and an agglomerated appearance with an average size of 197.40 nm. The GO/PEG/Bru-FA NCs treatment remarkably inhibited the growth of the tested pathogens. The findings of the MTT assay evidenced that the GO/PEG/Bru-FA NCs effectively reduced the HepG2 cell growth while not showing toxicity to the Vero cells. The findings of the fluorescent assay proved that the GO/PEG/Bru-FA NCs increased ROS generation, reduced MMP levels, and promoted apoptosis in the HepG2 cells. The levels of Bax, caspase-9, and -3 were increased, and Bcl-2 was reduced in the GO/PEG/Bru-FA NCs-treated HepG2 cells.
CONCLUSION: The results of this work demonstrate that GO/PEG/Bru-FA NCs suppress viability and induce apoptosis in HepG2 cells, indicating their potential as an anticancer candidate.