Serum Exosomes-Based Biomarker circ_0008928 Regulates Cisplatin Sensitivity, Tumor Progression, and Glycolysis Metabolism by miR-488/HK2 Axis in Cisplatin-Resistant Nonsmall Cell Lung Carcinoma.

Background: Nonsmall cell lung carcinoma (NSCLC) is a major cause of cancer-related death worldwide. The resistance of NSCLC to chemical drugs, such as cisplatin (CDDP), poses a heavy burden for NSCLC therapy. Herein, the effects of circular_0008928 (circ_0008928) on the CDDP sensitivity and biological behavior of CDDP-resistant NSCLC cells and underlying mechanism are revealed. Materials and Methods: The expression of circ_0008928 and microRNA-488 (miR-488) was detected by quantitative real-time polymerase chain reaction. The expression of hexokinase 2 (HK2) protein and exosome-specific proteins was determined by Western blot. The half-maximal inhibitory concentration (IC50) value of CDDP was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cell proliferation and migratory and invasive abilities were illustrated by cell counting kit-8 and transwell assays. Cell glycolysis metabolism was illustrated by extracellular acidification rate assay, glucose kit and lactate kit assays and Western blot analysis. The binding sites between miR-488 and circ_0008928 or HK2 were predicted by starbase or microT-CDS online database, and identified by dual-luciferase reporter and RNA immunoprecipitation assays. Results: Circ_0008928 expression and HK2 protein expression were significantly upregulated, while miR-488 expression was obviously downregulated in NSCLC cells and CDDP-resistant NSCLC cells. Circ_0008928 expression was increased in serum exosomes of CDDP-resistant NSCLC patients compared with CDDP-sensitive NSCLC patients. In addition, circ_0008928 silencing improved CDDP sensitivity and attenuated CDDP-induced cell proliferation, migration, invasion, and glycolysis metabolism. Circ_0008928 was a sponge of miR-488, and miR-488 bound to HK2 in CDDP-resistant NSCLC cells. Furthermore, both miR-488 inhibitor and HK2 overexpression attenuated circ_0008928 absence-mediated impacts on CDDP sensitivity and tumor process in CDDP-resistant NSCLC. Conclusions: Circ_0008928 knockdown improved CDDP sensitivity and hindered cell proliferation, migration, invasion, and glycolysis metabolism by miR-488/HK2 axis in CDDP-resistant NSCLC. This finding provides a new mechanism for studying CDDP-resistant therapy in NSCLC.

Role of FSCN1 in the tumor microenvironment of lung squamous cell carcinoma.

Tumor microenvironment (TME) regulated the development of the Lung squamous cell carcinoma (LUSC). To know more about the LUSC, this study tried to figure the role of fscin actin-bundling protein 1 (FSCN1) in the TME. We identified the FSCN1 as the hub immune gene in LUSC, with the use of weighted gene co-expression network analysis (WGCNA) and the Human Protein Atlas. Furthermore, we verified the higher expression of FSCN1 in LUSC compared with the normal tissues by quantitative reverse transcription PCR (qRT-PCR) and immunohistochemistry. We then explored the associations among FSCN1, immune infiltrations, and inflammatory factors with the use of Gene Expression Profiling Interactive Analysis (GEPIA) and Tumor IMmune Estimation Resource (TIMER). As a result, the expressions of FSCN1 was negatively related to the immune infiltrations, and positively related to the expressions of IL1A, IL1B, TGFB1 and TGFA. Moreover, we used the single-cell RNA-sequencing (scRNA-seq) data of LUSC to figure out the expressions level of FSCN1, IL1A, IL1B, TGFB1 and TGFA in the different cell type's of the TME. Finally, through the cytological experiments, we found that FSCN1 affected by TGFB1 contributes to the proliferation, anti-apoptotic effect, migration and invasion of the LUSC cells. In summary, this study Identified FSCN1 as the potential therapeutic target of LUSC, and reveals a complicated immune and inflammatory net in the TME.

Efficacy of flaxseed oil compared with fish oil supplementation in the treatment of coronary heart disease: a retrospective study.

Background: Many studies have demonstrated the beneficial effects of omega-3 fatty acids in animal models and human diseases. Compared with commonly used fish oil, flaxseed oil has better palatability. However, the relative efficacy of the two types of oil on the cardiovascular health of type 2 diabetes mellitus (T2DM) patients with coronary heart disease (CHD) is unclear. Methods: This was a retrospective study based on the prospectively maintained database of Hubei Provincial Hospital of Traditional Chinese Medicine. Patients meeting the inclusion criteria were enrolled and then divided into two groups: the flaxseed oil group received 1,000 mg flaxseed oil, which contains 400 mg of α-Linolenic acid, as their omega-3 fatty acid source; the fish oil group received 1,000 mg of fish oil, which contains 250 mg of eicosapentaenoic acid and 150 mg of docosahexaenoic acid. The primary outcome was cardiovascular risk biomarker changes between the two groups. P values less than 0.05 were considered statistically significant. Results: A total of 120 patients were enrolled in the present study: 60 in the flaxseed oil group and 60 in the fish oil group. After a median follow-up of 10.0 weeks (95% CI: 8.4-11.6 weeks), flaxseed oil was found to be significantly better at reducing serum insulin levels and high-sensitivity C-reactive protein (hs-CRP) levels than fish oil (P=0.03 and P=0.02, respectively). The effects of flaxseed oil and fish oil on homeostatic model assessment for insulin resistance (HOMA-IR), fasting plasma glucose (FPG); body weight, and body mass index (BMI) were found to be similar. Moreover, patients who received flaxseed oil tended to have a better overall survival than those who received fish oil, although the difference was not statistically significant (P=0.067). Conclusions: Compared with fish oil, flaxseed oil was more effective in reducing serum insulin levels and hs-CRP levels for T2DM patients with CHD. For these patients, flaxseed oil might become a novel choice.

[The effect of dexmedetomidine on autophagy and apoptosis in intestinal ischemia reperfusion-induced lung injury].

OBJECTIVE: To investigate the protective effects of dexmedetomidine against autophagy and apoptosis in intestinal ischemia reperfusion (I/R)-induced lung injury. METHODS: Twenty-four SD rats were randomly and equally divided into 4 groups according to the random number table: control group, I/R group, small dose of dexmedetomidine (D1) group and large dose of dexmedetomidine (D2) group. The model of lung injury was induced by occlusion of the superior mesenteric artery for 60 min followed by 12 h reperfusion. Rats in D1 and D2 groups received intravenous injection of dexmedetomidine at dose of 1 µg/kg and 5 µg/kg respectively. Rats in control and I/R groups were given same volume of normal saline. Pathological changes were detected by hematoxylin-eosin (HE) staining. The microtubule-associated protein 1 light chain 3(LC3)II/I ratio, Beclin-1, Bax and Bcl-2 expression were measured by Western blot. Cell apoptosis was assayed by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL), while caspase-3 activity was evaluated by immunohistochemistry. RESULTS: Significant infiltration of neutrophils and thickened alveolar walls were observed in the I/R group compared to the control group, which were improved by dexmedetomidine (5 µg/kg) treatment. Compared to the control group, the apoptosis cells [(69 ± 8) cells/field], LC3 II/I ratio(57 ± 8), Beclin-1(487%± 45%) and Bax (358% ± 37%) expression were markedly increased (P<0.05), while Bcl-2 (39% ± 5%) expression was decreased (P<0.05) in the I/R group. Compared to the I/R group, the apoptosis cells [(32 ± 5) cells/field], LC3 II/I ratio(27 ± 4), Beclin-1 (285%± 41%) and Bax (181% ± 25%) expression were markedly reduced (P<0.05), while Bcl-2 (91% ± 9%) expression was increased (P<0.05) in the D2 group. CONCLUSIONS: Autophagy and apoptosis were activated in intestinal I/R-induced lung injury. Dexmedetomidine ameliorated intestinal I/R-induced lung injury via reducing autophagy and apoptosis.

[5-aza-2'-deoxycytidine suppresses the growth of human lung adenocarcinoma cells in nude mouse xenograft models and its effect on methylation status and expression of TFPI-2 gene].

OBJECTIVE: To investigate the inhibitory effect of classic demethylating drug 5-aza-2'-deoxycytidine (5-Aza-CdR) on the growth of human lung adenocarcinoma cells in nude mouse xenograft models, and to observe its effect on methylation status and expression of TFPI-2 gene in the nude mouse xenograft tissues. METHODS: The nude mouse xenograft model was established by subcutaneous inoculation of human lung adenocarcinoma A549 cells. According to different doses of 5-Aza-CdR, the tumor-bearing nude mice were randomly divided into experimental groups (0.5 mg/kg group, 1 mg/kg group, 2 mg/kg group) and control group (0 mg/kg group). The tumor growth in the nude mice was observed. The methylation status and the expression of TFPI-2 gene mRNA and protein were detected by methylation specific polymerase chain reaction, real-time fluorescent quantitative polymerase chain reaction and Western blot assay. RESULTS: The nude mice were euthanized at 28 days after intraperitoneal injection of 5-Aza-CdR. The body weight of tumor-bearing nude mice was (27.12 ± 0.38) g in the 0 mg/kg group, (26.80 ± 0.18) g in the 0.5 mg/kg group, (26.67 ± 0.28) g in the 1 mg/kg group, and (26.50 ± 0.26) g in the 2 mg/kg group, showing no significant difference among them (P > 0.05). The volume of xenograft tumors in the 0 mg/kg group was (709.22 ± 2.87)mm³, (400.67 ± 2.68)mm³ in the 0.5 mg/kg group, (285.71 ± 2.91)mm³ in the 1 mg/kg group, and (230.44 ± 3.15)mm³ in the 2 mg/kg group, showing a significant difference (P < 0.05). There were complete methylation of TFPI-2 gene in the 0 mg/kg group, incomplete methylation in the 0.5 and 1 mg/kg groups, and unmethylation in the 2 mg/kg group. The relative mRNA level in the 0, 0.5, 1, 2 mg/kg groups were 1.00 ± 0.00, 1.67 ± 0.07, 3.40 ± 0.24, and 5.55 ± 0.61, respectively (P < 0.05). The relative expression level of TFPI-2 protein in the 0, 0.5, 1, 2 mg/kg groups was 0.18 ± 0.02, 0.36 ± 0.01, 0.64 ± 0.02, and 0.81 ± 0.20, respectively (P < 0.05). CONCLUSIONS: 5-Aza-CdR suppresses the tumor growth of human lung adenocarcinoma cells in nude mouse xenograft models, and induces expression of TFPI-2 gene in the xenograft tumor cells. The mechanism might be that 5-Aza-CdR induces re-expression of demethylated TFPI-2 gene by demethylation, and thus inhibits the growth and proliferation of human lung adenocarcinoma cells.

Surgical treatment of an invasive thymoma extending into the superior vena cava and right atrium.

Although invasive thymoma commonly infiltrates neighbouring mediastinal structures, its extension into the superior vena cava (SVC) and consequent SVC occlusion are rare. In such cases, the urgent removal of the thymoma and radical resection of the infiltrated SVC representreasonable options, since induction therapy is time-consuming and useless for symptom resolution. A case of invasive thymoma extending into the SVC and right atrium (RA) with SVC syndrome is reported. The patient underwent a combined resection of the invasive tumor and SVC under cardiopulmonary bypass (CPB), and the SVC and bilateral brachiocephalic vein (BCV) were reconstructed with an autologous pericardial 'Y' conduit. After 40 months of follow-up, the patient showed a patent graft and no tumor recurrence.

Effect of 5-aza-2'-deoxycytidine on cell proliferation of non- small cell lung cancer cell line A549 cells and expression of the TFPI-2 gene.

OBJECTIVE: The present study employed 5-aza-2'-deoxycytidine (5-Aza-CdR) to treat non-small cell lung cancer (NSCLC) cell line A549 to investigate the effects on proliferation and expression of the TFPI-2 gene. METHODS: Proliferation was assessed by MTT assay after A549 cells were treated with 0, 1, 5, 10 µmol/L 5-Aza-CdR, a specific demethylating agent, for 24 ,48 and 72h. At the last time point cells were also analyzed by flow cytometry (FCM) to identify any change in their cell cycle profiles. Methylation-specific polymerase chain reaction (MSPCR), real time polymerase chain reaction(real-time PCR) and western blotting were carried out to determine TFPI-2 gene methylation status, mRNA expression and protein expression. RESULTS: MTT assay showed that the growth of A549 cells which were treated with 5-Aza-CdR was significantly suppressed as compared with the control group (0 µmol/L 5-Aza-CdR). After treatment with 0, 1, 5, 10 µmol/L 5-Aza-CdR for 72h, FCM showed their proportion in G0/G1 was 69.7±0.99%, 76.1±0.83%, 83.8±0.35%, 95.5±0.55% respectively (P<0.05), and the proportion in S was 29.8±0.43%, 23.7±0.96%, 15.7±0.75%, 1.73±0.45%, respectively (P<0.05), suggesting 5-Aza-CdR treatment induced G0/G1 phase arrest. MSPCR showed that hypermethylation in the promoter region of TFPI-2 gene was detected in control group (0 µmol/L 5-Aza-CdR), and demethylation appeared after treatment with 1, 5, 10 µmol/L 5-Aza-CdR for 72h. Real-time PCR showed that the expression levels of TFPI-2 gene mRNA were 1±0, 1.49±0.14, 1.86±0.09 and 5.80±0.15 (P<0.05) respectively. Western blotting analysis showed the relative expression levels of TFPI-2 protein were 0.12±0.01, 0.23±0.02, 0.31±0.02, 0.62±0.03 (P<0.05). TFPI-2 protein expression in A549 cells was gradually increased significantly with increase in the 5-Aza-CdR concentration. CONCLUSIONS: TFPI-2 gene promoter methylation results in the loss of TFPI-2 mRNA and protein expression in the non-small cell lung cancer cell line A549, and 5-Aza-CdR treatment could induce the demethylation of TFPI-2 gene promoter and restore TFPI-2 gene expression. These findings provide theoretic evidence for clinical treatment of advanced non-small cell lung cancer with the demethylation agent 5-Aza-CdR. TFPI-2 may be one molecular marker for effective treatment of advanced non-small cell lung cancer with 5-Aza-CdR.