ACTN4 and the pathways associated with cell motility and adhesion contribute to the process of lung cancer metastasis to the brain.

BACKGROUND: The aim of this study was to identify critical gene pathways that are associated with lung cancer metastasis to the brain. METHODS: The RNA-Seq approach was used to establish the expression profiles of a primary lung cancer, adjacent benign tissue, and metastatic brain tumor from a single patient. The expression profiles of these three types of tissues were compared to define differentially expressed genes, followed by serial-cluster analysis, gene ontology analysis, pathway analysis, and knowledge-driven network analysis. Reverse transcription-polymerase chain reaction (RT-PCR) was used to validate the expression of essential candidate genes in tissues from ten additional patients. RESULTS: Differential gene expression among these three types of tissues was classified into multiple clusters according to the patterns of their alterations. Further bioinformatic analysis of these expression profile data showed that the network of the signal transduction pathways related to actin cytoskeleton reorganization, cell migration, and adhesion was associated with lung cancer metastasis to the brain. The expression of ACTN4 (actinin, alpha 4), a cytoskeleton protein gene essential for cytoskeleton organization and cell motility, was significantly elevated in the metastatic brain tumor but not in the primary lung cancer tissue. CONCLUSIONS: The signaling pathways involved in the regulation of cytoskeleton reorganization, cell motility, and focal adhesion play a role in the process of lung cancer metastasis to the brain. The contribution of ACTN4 to the process of lung cancer metastasis to the brain could be mainly through regulation of actin cytoskeleton reorganization, cell motility, and focal adhesion.

Tracheal replacement with aortic grafts: Bench to clinical practice.

Tracheal reconstruction following extensive resection for malignant or benign lesions remains a major challenge in thoracic surgery. Numerous studies have attempted to identify the optimal tracheal replacement with different biological or prosthetic materials, such as various homologous and autologous tissues, with no encouraging outcomes. Recently, a few clinical studies reported attaining favorable outcomes using in vitro or stem cell-based airway engineering and also with tracheal allograft implantation following heterotopic revascularization. However, none of the relevant studies offered a standardized technology for airway replacement. In 1997, a novel approach to airway reconstruction was proposed, which involved using aortic grafts as the biological matrix. Studies on animal models reported achieving in-vivo cartilage and epithelial regeneration using this approach. These encouraging results inspired the subsequent application of cryopreserved aortic allografts in humans for the first time. Cryopreserved aortic allografts offered further advantages, such as easy availability in tissue banks and no requirement for immunosuppressive treatments. Currently, stented aortic matrix-based airway replacement has emerged as a standard approach, and its effectiveness was also verified in the recently reported TRITON-01 study. In this context, the present review aims to summarize the current status of the application of aortic grafts in tracheal replacement, including the latest advancements in experimental and clinical practice.

Targeted therapy of RET fusion-positive non-small cell lung cancer.

Lung cancer has very high morbidity and mortality worldwide, and the prognosis is not optimistic. Previous treatments for non-small cell lung cancer (NSCLC) have limited efficacy, and targeted drugs for some gene mutations have been used in NSCLC with considerable efficacy. The RET proto-oncogene is located on the long arm of chromosome 10 with a length of 60,000 bp, and the expression of RET gene affects cell survival, proliferation, growth and differentiation. This review will describe the basic characteristics and common fusion methods of RET genes; analyze the advantages and disadvantages of different RET fusion detection methods; summarize and discuss the recent application of non-selective and selective RET fusion-positive inhibitors, such as Vandetanib, Selpercatinib, Pralsetinib and Alectinib; discuss the mechanism and coping strategies of resistance to RET fusion-positive inhibitors.

Tumor-derived exosomal circRNA_102481 contributes to EGFR-TKIs resistance via the miR-30a-5p/ROR1 axis in non-small cell lung cancer.

Exosomes are messengers for intercellular communication and signal transduction. Circular RNA (circRNA) abnormal expression and regulation are involved in the occurrence and development of a variety of tumors. In the present study, exosomes in the serum of five patients with non-small cell lung cancer (NSCLC) were isolated before and after EGFR-TKIs resistance, and the circRNA expression profile was screened using a circRNA microarray. The effects of the exosome circRNA_102481 on cell proliferation and apoptosis were analyzed. The interaction between miR-30a-5p and circRNA_102481 or ROR1 was predicted by starBase software, and was confirmed by RNA pull-down and dual-luciferase reporter assays. The results showed that exosomes containing circRNA_102481 were significantly up-regulated in NSCLC with EGFR-TKIs resistance (p<0.05), and that circRNA_102481 was mainly secreted by EGFR-TKIs resistance cell via exosomes (p<0.05). Both circRNA_102481 silencing and si-circRNA_102481 transported by exosomes could inhibit EGFR-TKIs resistance cell proliferation and promote cell apoptosis and circRNA_102481 overexpression could promote EGFR-TKIs sensitive cell proliferation and inhibit cell apoptosis in vitro (p<0.05). CircRNA_102481 served as a miR-30a-5p sponge to regulate ROR1 expression (p<0.05). Furthermore, the expression of circRNA_102481 in exosomes was associated with TNM stage, tumor differentiation status, brain metastasis, and PFS and OS duration. Therefore, it was concluded that tumor-derived exosomal circRNA_ 102481 could contribute to EGFR-TKIs resistance via the microRNA-30a-5p/ROR1 axis in NSCLC. Exosomal circRNA_102481 may serve as a novel diagnostic biomarker and a therapeutic target for EGFR-TKIs resistance in NSCLC.

Epidermal growth factor receptor (EGFR) mutation and p-EGFR expression in resected non-small cell lung cancer.

Lung cancer, specifically non-small cell lung cancer (NSCLC), is a leading cause of mortality worldwide. In China, a dramatic increase in the incidence of NSCLC is expected in the next 20 years (Molina et al. Mayo Clin Proc. 2008;83:584­594). Mutated epidermal growth factor receptor (EGFR) status is a known predictor of response to tyrosine kinase inhibitors (TKIs), and immunohistochemistry may be a less costly way of predicting presence of mutation. In this study, mutation analysis of EGFR in 218 cases of NSCLC was performed. One hundred thirty tissue samples were examined via immunohistochemistry of p-EGFR (Y1045 and Y1068) and correlated with mutation status. Mutations were seen in 29% of patients, and were correlated with female sex, nonsmoking history, and adenocarcinoma histology. Phosphorylation at Y1045 was noted in 52% of cases, but in 71% of cases with EGFR mutation (P = .003). Phosphorylation of Y1068 was seen in 55% of cases but in 73% of cases with EGFR mutation (P = .006). This study correlating EGFR mutation with p-EGFR expression in resected NSCLC is one of the largest to date, although TKI response could not be assessed. The data show that, among Chinese patients, detection of p-1045 and p-1068 expression with immunohistochemistry predicts EGFR mutations. Immunohistochemical analysis of p-EGFR may be useful to predict responses to TKI therapy, although future studies are necessary.

[A comparative study of survival time of surgery combined with chemotherapy and non-surgical chemotherapy in SCLC].

OBJECTIVE: To compare the survival time of surgery combine with chemotherapy with non-surgery chemotherapy in SCLC. METHODS: 37 SCLC post operation patients were received 4-6 cycles of chemotherapy of "etoposide + cisplatin or carboplatin". 37 SCLC patients were selected be control group, whose age, stage and site of tumor were similar to surgery combine with chemotherapy group, and received 6 cycles of chemotherapy of "etoposide + cisplatin or carboplatin". Two groups were followed up 5 year and compared with the survival time. RESULTS: The 1, 3, 5 year survival rate of surgery combined with chemotherapy group were 72.97%, 35.13%, 21.62%. The 1, 3, 5 year survival rate of control group were 54.05%, 13.51%, 5.41%. The survival rate of stage I (P = 0.01) was significantly different, but stage II (P = 0.06) and stage III (P = 0.836) were no difference between the two group of SCLC. CONCLUSION: It was found that surgery combined with chemotherapy got satisfactory effects in stage I SCLC.

A study on changes and clinical significance of blood glucose, blood lipid and inflammation in patients with ovarian cancer.

PURPOSE: To investigate the changes in blood glucose, blood lipid and inflammation in patients with ovarian cancer and their clinical significance. METHODS: 67 patients diagnosed with ovarian cancer and treated in our hospital from January 2018 to December 2018 constituted the observation group. Fifty healthy women in the corresponding time period were enrolled as the control group. The levels of blood glucose, blood lipid and inflammation were compared between the two groups, and then the changes in those levels in ovarian cancer patients in different clinical stages were analyzed. RESULTS: The levels of fasting blood glucose and 2-h postprandial blood glucose of the observation group were significantly higher than those of the control group, and they were also significantly higher in patients in stage III-IV than those in stage I-II (p<0.05). In the observation group, the levels of total cholesterol (TC) and high-density lipoprotein cholesterol (HDL-C) were lower than those in the control group. Compared with the patients in stage I-II, the patients in stage III-IV had remarkably lower levels of TC and HDL-C (p<0.05). The levels of C-reactive protein (CRP), tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in the observation group were evidently higher than those in the control group, and they were markedly higher in patients in stage III-IV than those in stage I-II (p<0.05). CONCLUSIONS: Both the metabolism disorders of blood lipid and blood glucose and inflammatory response are more obvious in ovarian cancer patients, indicating that these indicators have a place for early screening and clinical treatment of ovarian cancer.

Overexpression of circ-0067934 is associated with increased cellular proliferation and the prognosis of non-small cell lung cancer.

Previous studies have suggested that circular RNAs (circRNAs) serve crucial functions in various human diseases, including cancer. The results of the present study demonstrated that the expression levels of circ-0067934 were significantly increased in non-small cell lung cancer (NSCLC) tissues and NSCLC cells compared with that in adjacent normal tissues and a normal human bronchial epithelium cell line, as assessed using reverse transcription-quantitative polymerase chain reaction. Increased expression levels of circ-0067934 were associated with lymph node metastasis and advanced tumor stage in patients with NSCLC. Increased levels of circ-0067934 were significantly associated with the poor overall survival (OS) time of patients with NSCLC compared with low circ-0067934 expression levels, as assessed using Kaplan-Meier estimator survival curves and log-rank tests. Additionally, circ-0067934 was identified as an independent risk factor for OS time in patients with NSCLC, as assessed using univariate and multivariate Cox proportional hazards regression models. Additionally, cell proliferation and colony formation assays were employed to assess cell proliferation. The results demonstrated that the proliferative ability was significantly inhibited when circ-0067934 was knocked down in NSCLC cells. Therefore, these results indicated that circ-0067934 may be a predictive marker for the prognosis of NSCLC and a target for the treatment of the disease.

Overexpression of CD44 is associated with the occurrence and migration of non-small cell lung cancer.

Non-small cell lung cancer (NSCLC) is a potentially fatal disease and the incidence is increasing annually. In order to diagnose and treat NSCLC effectively, greater understanding of its molecular mechanism is required. In the present study, 36 NSCLC tissues and 10 normal tissues were selected. Reverse transcription­quantitative polymerase chain reaction (RT­qPCR) was used to analyze the CD44 mRNA expression level in NSCLC tissue and DNA sequencing was performed to further verify the CD44 expression level. Differentially expressed genes between tumor tissues and controls were determined by DNA sequencing and the Gene_act_net between CD44 and its associated genes was constructed. Gene Ontology (GO) term enrichment analysis of the differentially expressed genes was performed by the Biological Networks Gene Ontology tool. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was performed based on the Expression Analysis Systematic Explorer test applied in the Database for Annotation, Visualization and Integrated Discovery. RT­qPCR results showed that CD34 was overexpressed in 21 of the 36 NSCLC tissues (58.3%). The Gene_act_net indicated that there were 20 differentially expressed genes with 17 upregulated and 3 downregulated. Among them, CD44, MET, ERBB2, EGFR, AKT1, IQGAP1 and STAT3 were associated with the occurrence and migration of NSCLC. In KEGG pathway analysis, extracellular matrix­receptor interaction and hematopoietic cell lineage pathways were the most affected by overexpressed CD44; and thus may be important in the development and migration of NSCLC. In conclusion, CD44 was overexpressed in NSCLC and the overexpression was associated with the occurrence of NSCLC and migration of NSCLC cells.

5-Aza-CdR can reverse gefitinib resistance caused by DAPK gene promoter methylation in lung adenocarcinoma cells.

To explore the relationship between death associated protein kinase (DAPK) gene promoter methylation and gefitinib resistance in Lung adenocarcinoma cell lines. EGFR-mutation lung adenocarcinoma cell lines PC9 and the gefitinib-resistant with T790M Mutation cell lines PC9/GR were chosen as cell models, and PC9/GR were treated with 5-aza-CdR (1 µmol/L). The experiments were divided into three groups: PC9 group, PC9/GR group and PC9/GR with 5-Aza-CdR pretreatment group. Treat three groups cell with different concentrations gefitinib, the cell proliferation was determined by MTT assay. The apoptotic rates were detected by flow cytometry. The methylation of DAPK gene promoter region was examined by methylation-specific PCR (MSP). The expressions of DAPK protein were detected by Western blot. MTT results showed that the half maximal inhibitory concentration (IC50) of PC9 and PC9/GR cell lines increase from 0.12 µmol/L to 8.52 µmol/L. But after treated with 5-aza-CdR, the IC50 of PC9/GR cell lines decrease to 4.35 µmol/L, and the resistance index (RI) decrease from 71 to 36 (P<0.05). Flow cytometry results showed that the apoptosis rate were 24.80% ± 0.28%, 12.70% ± 0.31%, 19.8% ± 0.15% respectively. MSP results showed that DAPK gene promoter region was un-methylated in PC9 cells and methylated in PC9/GR cells, when treated with 5-aza-CdR, DAPK gene promoter region was partly methylated in PC9/GR cells (P<0.05). Western blot results showed that the levels of DAPK protein were reduced significantly in PC9/GR cell lines compared with PC9, and after treated with 5-aza-CdR, the expression levels of DAPK protein in PC9/GR were increased (P<0.05). In conclusion, DAPK gene promoter methylation may contribute to the downregulation of DAPK gene and protein, and consequently affect the sensitivity of gefitinib in lung adenocarcinoma lines, induced gefitinib resistance. But 5-Aza-CdR can reverse gefitinib resistance by demethylation of DAPK gene promoter.

Clinicopathologic significance of ß-catenin and matrix metalloproteinase-2 expression in non-small cell lung cancer.

The purpose of this study was to analyze ß-catenin and matrix metalloproteinase-2 (MMP-2) expression in non-small cell lung cancer (NSCLC) and to investigate the association between their expression and clinicopathologic characteristics of NSCLC patients. Immunohistochemistry was performed to examine ß-catenin and MMP-2 protein expression in 39 resected NSCLC samples and 8 adjacent normal lung tissues. Statistical analysis with SPSS13.0 software was performed to investigate the association between ß-catenin and MMP-2 expression and clinicopathologic features of the patients. Expression of cytosolic ß-catenin in NSCLC tissue was significantly higher than that in normal tissues (P < 0.001). In addition, cytosolic protein expression of ß-catenin in lung squamous cell carcinoma was significantly elevated compared to that in lung adenocarcinoma (P = 0.02). However, cell membrane protein expression of ß-catenin in squamous cell carcinoma was lower than that in adenocarcinoma (P = 0.041). Cytosolic MMP-2 protein expression in NSCLC samples was significantly higher than that in normal tissues (P = 0.002). MMP-2 expression in N (1-2) NSCLC patients was significantly increased relative to N (0) patients (P = 0.019). However, statistical analysis showed no correlation between ß-catenin and MMP-2 expression in NSCLC samples. Collectively, our results show that cytosolic protein expression of ß-catenin in NSCLC samples is increased relative to normal lung tissues. Also, expression of ß-catenin is significantly elevated in squamous cell carcinoma compared to that in lung adenocarcinoma subtypes. Additionally, MMP-2 expression in N (1-2) NSCLC tissues is higher than that in N (0) lung tissue. There is no correlation between ß-catenin and MMP-2 expression in NSCLC, and our study suggests that evaluation of ß-catenin and MMP-2 expression may have potential in diagnosis and progression in patients with NSCLC.