Atorvastatin suppressed proliferation and facilitated apoptosis of A549 cells through mediating recruitment of Fas and CD59 in lipid raft

Purpose: Lipid raft facilitated progression of NSCLC and atorvastatin could break cholesterol. The purpose of this study was to determine the potential mechanism of atorvastatin through lipid raft mediation in NSCLC.Methods: A549 cells were first grouped as NC, methyl-β-cyclodextrin (MβCD, lipid raft inhibitor, 5mM and 10mM), atorvastatin (0, 5mM, 10mM and 15mM) and MβCD (10mM) with atorvastatin (15mM). Later 10mM MβCD treated A549 cells were divided into three groups: NC, 3BDO (mTOR agonist) (60μM) and 3BDO (60μM) with atorvastatin (15mM) group. Thereafter, FLOT-2, SLP-2 expressions were assessed with RT-qPCR，mTOR proteins were measured by western blot and cell viability by CCK-8 method. Meanwhile, apoptosis was analyzed by flow cytometry. Moreover, lipid raft isolation was performed for acquiring Fas and CD59 and concentrations were detected by ELISA.Results: MβCD treatment significantly inhibited FLOT-2 and SLP-2 RNA expressions and cell viability of A549 cells but up-regulated apoptosis. Besides that, Fas protein level was promoted and CD59 was suppressed. Atorvastatin also repressed FLOT-2 and SLP-2 RNA levels. Meanwhile, atorvastatin downregulated cell viability and accelerated apoptosis. Moreover, Fas was increased and CD59 was inhibited by atorvastatin and MβCD enhanced functions of atorvastatin. MβCD inhibited mTOR RNA expression in A549 cells was increased by 3BDO but atorvastatin restored 3BDO caused up-regulation of mTOR. Furthermore, up regulated cell viability of A549 by 3BDO was declined with atorvastatin and decreased apoptosis by 3BDO was reversed through atorvastatin. Fas suppressed by 3BDO and promotion of CD59 were also resumed by atorvastatin.Conclusion: Atorvastatin suppressed FLOT-2 and SLP-2 and mediated recruitments of Fas and CD59 with suppressing cell viability and facilitating apoptosis of A549 cells via mTOR signaling pathway

Analysis of Differential Gel Electrophoresis of Paclitaxol Resistant and Sensitive Lung Adenocarcinoma Cells' Secretome

Background and objective Paclitaxol (PTX) resistance is one of main factors which affect the outcome of chemotherapy of lung adenocarcinoma. The aim of this study is to compare the secreted protein expression profiles between Paclitaxol (PTX) resistant and sensitive lung adenocarcinoma cells by proteomic research method, so as to provide evidence of choosing individual chemotherapy drugs in clinical treatment. Methods Total secreted proteins extracted from a PTX sensitive cell line A549 and a PTX resistant cell line A549-Taxol were separated by fluorscent differential gel electrophoresis (DIGE). High quality 2-DE profiles were obtained and analyzed by Decyder 6.5 analysis software to screen differentially expressed protein spots. Those spots were identified by mass spectrometry. Results 2-DE patterns of lung adenocarcinoma cells with high-resolution and reproducibility were obtained. 76 significantly differentially expressed protein spots were screened, 19 proteins were identified by mass spectrometry. The identified proteins could be classified into different catogories: metabolic enzyme, extracellular matrix (ECM) degradation enzyme, cytokine, signal transducer, cell adhesion, and so on. Conclusion Multiple secreted proteins related to chemoresistance of A549-Taxol cells were identified in this study for the first time. The results presented here would provide clues to identify new serologic chemoresistant biomarkers of NSCLC

A retrospective study for prognostic significance of type II diabetes mellitus and hemoglobin A1c levels in non-small cell lung cancer patients treated with pembrolizumab

Background: Diabetes mellitus (DM) is common and recognized as a risk factor for developing non-small cell lung cancer (NSCLC) while the prognostic evaluation is still controversial. As immunotherapy is widely used in clinical practice, its efficacy and survival should be investigated in patients with DM. Methods: We retrospectively recruited 266 locally advanced and metastatic NSCLC patients who received pembrolizumab alone or in combination with chemotherapy. Patients\u27 clinicopathological data, including age, history of DM, hemoglobin A1c (HbA1c), genetic tumor profiling, and survival data were collected. Associations between clinical characteristics and survival were evaluated by univariate and multivariate analyses. Results: In this cohort, 15.04 % (40/266) of the patients had a history of DM. Fifty-nine (22.2 %) patients had a HbA1c level ≥ 6.5 %. A total of 169 (63.5 %) patients received 1st-line therapy, and 97 (36.5 %) received 2nd- or subsequent-line therapy. Patients with high ( ≥ 6.5 %) HbA1c and lower ( \u3c 35 g/L) albumin levels at baseline had worse survivals, and epidermal growth factor receptor (EGFR) mutants significantly associated with worse outcomes at normal HbA1c ( \u3c 6.5%) levels (all P \u3c 0.05). Among the 1st-line therapy patients, a higher HbA1c level ( ≥ 6.5 %) at baseline indicated a worse overall survival (OS) (2-year survival rate: 31.25 % vs. 27.03 %, P = 0.045), tumor protein p53 (TP53) alternations and high programmed death-ligand 1 (PD-L1) expression ( ≥ 50 %) were significantly associated with better outcomes (P \u3c 0.05). For 2nd- or subsequent-line patients, EGFR mutants and non-squamous carcinomas (non-SCs) indicated worse survivals, and the normal peripheral blood markers of the carcinoembryonic antigen (CEA), C-reactive protein (CRP), albumin levels were favorable prognostic factors for survivals. In non-SCs, Kirsten rat sarcoma viral oncogene homolog (KRAS) mutations, high PD-L1 expression, and normal alkaline phosphatase (ALP) levels favored better progression-free survival (PFS), while EGFR mutants indicated poor PFS (P \u3c 0.05). Conclusions: Among patients treated with 1st-line immunotherapy, a higher HbA1c level ( ≥ 6.5 %) indicated dismal OS, while history of DM, baseline blood glucose levels, and glucose changes during the treatment process were not significantly associated with any of the outcomes

A columnar regular-porous stainless steel reaction support with high superficial area for hydrogen production

To improve hydrogen production (HP) performance of regular-porous structure (RPS), a columnar RPS with small specific surface area and high superficial area is developed. A numerical simulation model of regular-porous stainless steel structure (RPSSS) is established. Subsequently, heat transfer performance, pressure loss, temperature, methanol concentration, H2 concentration distributions and HP performance of the columnar RPSSS with small specific surface area and high superficial area and the body-centered cubic RPSSS with high specific surface area and small superficial area are compared. Then, temperature, methanol concentration, H2 concentration distributions and HP performance of axial and longitudinal size-enlarged columnar RPSSSs are studied. The results show that compared to the body-centered cubic RPSSS, the columnar RPSSS has higher methanol conversion, larger H2 flow rate and higher CO selectivity. Especially in the condition of 300 °C wall temperature and 12 mL/h methanol-water mixture injection rate (MWMIR), the methanol conversion, H2 flow rate and CO selectivity of the columnar RPSSS are increased by 12.3%, 9.24% and 30%, respectively, indicating that the superficial area of RPSSS is more important for its HP performance compared to its specific surface area. Compared to the longitudinal size-enlarged columnar RPSSS, the axial size-enlarged columnar RPSSS has higher methanol conversion, larger H2 flow rate and higher CO selectivity. This research work provides a new method for the optimization of hydrogen production reaction support (HPRS)

Novel nickel foam with multiple microchannels as combustion reaction support for the self-heating methanol steam reforming microreactor

To improve hydrogen production performance of self-heating methanol steam reforming (MSR) microreactor, novel nickel foam with multiple microchannels was proposed as combustion reaction support. A wall temperature comparison of the methanol combustion microreactors with nickel foam catalyst support and particles catalyst support in the combustion reaction process was performed. According to the numerical simulation result of combustion reaction of nickel foam, the shape and size of multiple microchannels of nickel foam were determined. The laser processing was then used to fabricate the multiple microchannels of nickel foam. The experimental results show that the methanol combustion microreactor with nickel foam loaded with Pt catalyst exhibits similar wall temperature distribution with the methanol combustion microreactor with Pt/γ-Al2O3 particles reaction support. Compared with the nickel foam without a microchannel, the maximum temperature difference (ΔTmax) and the maximum temperature of nickel foam with multiple microchannels were decreased, respectively, by 57.8% and 33.8 °C when 1.1 mL/min methanol flow rate was used. Hydrogen production performance of the self-heating MSR microreactor using the nickel foam with multiple microchannels increased by about 21% when 430 °C reforming temperature and 4 mL/h methanol–water mixture flow rate were performed

Structural design of self-thermal methanol steam reforming microreactor with porous combustion reaction support for hydrogen production

To replace the traditional electric heating mode and increase methanol steam reforming reaction performance in hydrogen production, methanol catalytic combustion was proposed as heat-supply mode for methanol steam reforming microreactor. In this study, the methanol catalytic combustion microreactor and self-thermal methanol steam reforming microreactor for hydrogen production were developed. Furthermore, the catalytic combustion reaction supports with different structures were designed. It was found that the developed self-thermal methanol steam reforming microreactor had better reaction performance. Compared with A-type, the △Tmax of C-type porous reaction support was decreased by 24.4 °C under 1.3 mL/min methanol injection rate. Moreover, methanol conversion and H2 flow rate of the self-thermal methanol steam reforming microreactor with C-type porous reaction support were increased by 15.2% under 10 mL/h methanol-water mixture injection rate and 340 °C self-thermal temperature. Meanwhile, the CO selectivity was decreased by 4.1%. This work provides a new structural design of the self-thermal methanol steam reforming microreactor for hydrogen production for the fuel cell

Detection of Genetic Mutations by Next-Generation Sequencing for Predicting Prognosis of Extensive-Stage Small-Cell Lung Cancer

Some studies have revealed that specific genetic mutations could be associated with chemotherapy response or even survival in small-cell lung cancer (SCLC). Our retrospective study aimed to identify the correlation between genetic mutations and progression-free survival (PFS) in extensive-stage SCLC after first-line chemotherapy. A total of 75 patients with extensive-stage SCLC confirmed by histopathology from February 2018 to February 2019 were retrospectively analyzed. The biopsy specimens of all patients were analyzed by Next-Generation Sequencing (NGS). All patients received first-line chemotherapy and follow-up at Shanghai Chest Hospital. Eleven genes were mutated in, at least, 10% of the 75 patients, including TP53 (96%), RB1 (77%), SMAD4 (32%), NOTCH1 (21%), PTEN (16%), FGFR1 (16%), KDR (15%), PIK3CA (15%), ROS1 (15%), BRCA2 (13%), and ERBB4 (10%). The median number of mutated genes among all patients was 5. Patients with more than 5 mutated genes (PFS = 6.7 months, P=0.004), mutant TP53 (PFS = 5.0 months, P=0.011), and mutant BRCA2 (PFS = 6.7 months, P=0.046) had better PFS after first-line chemotherapy than other patients. Multivariate Cox regression analysis showed that patients who achieved a PR (HR 3.729, 95% CI 2.038–6.822), had more than 5 mutated genes (HR 1.929, 95% CI 1.096–3.396), had BRCA2 mutations (HR 4.581, 95% CI 1.721–12.195), and had no liver metastasis (HR 0.415, 95% CI 0.181–0.951) showed improvements in PFS after first-line chemotherapy. In conclusion, the number of mutated genes and BRCA2 mutation status in extensive-stage SCLC were significantly related to PFS after first-line chemotherapy