Background: Hepcidin antimicrobial peptide (HAMP) is a small peptide hormone recognized for its role in iron metabolism and cancer treatment. The purpose of this study was to examine the influence of HAMP in NSCLC. Methods: The profile of NSCLC cells and tissues was characterized via HAMP. Gain- or loss-of-function cell models of HAMP were constructed, and CCK8, colony formation, and Transwell analyses were used to confirm the influence of HAMP on NSCLC cells. Upstream and downstream HAMP mechanisms in NSCLC were also analysed. Dual-luciferase reporter and pull-down assays confirmed the associations of miR-873-5p with HAMP, miR-873-5p, and the lncRNA KCNQ1OT1/SNHG14/XIST. Moreover, a xenograft model was established in nude mice for confirming the role of HAMP in NSCLC cell growth. Results: In addition, HAMP expression increased in NSCLC cells and tissues. In terms of cellular functions, the HAMP-overexpressing group exhibited elevated NSCLC cell proliferation, invasion, and migration. HAMP knockdown reversed these changes. Bioinformatics analysis indicated that miR-873-5p targeted HAMP, which affected the nuclear factor kappa B (NF-κB) pathway in NSCLC. HAMP activated the NF-κB pathway, which was negatively modulated by miR-873-5p. NF-κB inhibitor JSH-23 can partly suppress the proliferation, invasion, and migration in HAMP-overexpressed cells. Moreover, miR-873-5p was the target miRNA of long noncoding RNAs (lncRNAs), which included KCNQ1OT1, SNHG14, and XIST, and these three lncRNAs promoted HAMP. Conclusion: Noncoding RNA-mediated HAMP promotes NSCLC cell proliferation, migration, and invasion by initiating the NF-κB pathway.

Due to the limitations of traditional laboratory methods (TMs), identification of causative pathogens of numerous pulmonary infections (PIs) remains difficult. This study evaluated the value of metagenomic next generation sequencing (mNGS) in the identification of various respiratory pathogens. A total of 207 patients with TMs and mNGS data were collected for this retrospective study. TMs included sputum culture, blood, and bronchoalveolar lavage fluid (BALF) analysis, or polymerase chain reaction analysis of throat swabs. Otherwise, BALF was collected and analyzed using mNGS. For bacterial pathogens, sensitivities of mNGS as compared to TMs were 76.74% and 58.14% (P=0.012). For fungal pathogens, the detection rate of mNGS sensitivity was higher as compared to that of TMs (93.68% vs 22.11%; P<0.001). The positive predictive value and negative predictive value were also greater for mNGS. Use of mNGS for BALF analysis offers good specificity and thus facilitates to the clinical diagnosis of PIs.

Non-small cell lung cancer (NSCLC) encompasses approximately 85% of all lung cancer cases and is the foremost cancer type worldwide; it is prevalent in both sexes and known for its high fatality rate. Expanding scientific inquiry underscores the indispensability of microRNAs in NSCLC. Here, we probed the impact of miR-873-5p on NSCLC development and chemoresistance. qRT‒PCR was used to measure the miR-873-5p level in NSCLC cells with or without chemoresistance. A model of miR-873-5p overexpression was constructed. The proliferation and viability of NSCLC cells were evaluated through CCK8 and colony formation experiments. Cell migration and invasion were monitored via Transwell assays. Western blotting was used to determine the levels of YWHAE, PI3K, AKT, EMT, apoptosis, and autophagy-related proteins. The sensitivity of NSCLC cells to the chemotherapeutic agent gefitinib was assessed. Additionally, the correlation of YWHAE with miR-873-5p was validated via a dual-luciferase reporter assay and RNA immunoprecipitation (RIP). Overexpressed miR-873-5p suppressed migration, proliferation, invasion, and EMT while concurrently stimulating apoptotic processes. miR-873-5p was downregulated in NSCLC cells resistant to gefitinib. Upregulating miR-873-5p reversed gefitinib resistance by inducing autophagy. YWHAE was confirmed to be a downstream target of miR-873-5p. YWHAE overexpression promoted the malignant behaviors of NSCLC cells and boosted tumor growth, while these effects were reversed following miR-873-5p overexpression. Subsequent investigations revealed that overexpressing YWHAE promoted PI3K/AKT pathway activation, with miR-873-5p displaying inhibitory effects on the YWHAE-mediated PI3K/AKT signaling cascade. miR-873-5p affects proliferation, invasion, migration, EMT, autophagy, and chemoresistance in NSCLC by controlling the YWHAE/PI3K/AKT axis.

BACKGROUND:MicroRNAs (miRNAs) of plasma-derived small extracellular vesicles (sEVs) have been proven to be associated with metastasis in several types of cancer. This study aimed to detect miRNAs of plasma-derived sEVs as potential biomarkers for metastatic non-small cell lung cancer (NSCLC).METHODS:We assessed the miRNA profiles of plasma-derived sEVs from healthy individuals as the control group (CT group), NSCLC patients without distant organ metastasis as the NM-NSCLC group and patients with distant organ metastasis as the M-NSCLC group. Next-generation sequencing (NGS) was performed on samples, and differentially expressed miRNAs (DEMs) of the three groups were screened. Kyoto Encyclopedia of Genes and Genomes (KEGG) and ClueGO were used to predict potential pathways of DEMs. MiRNA enrichment analysis and annotation tool (miEAA) was used to understand changes in the tumour microenvironment in NSCLC. Quantitative reverse transcription polymerase chain reaction (qRT‒PCR) analysis was used to validate target miRNAs.RESULT:NGS was performed on 38 samples of miRNAs of plasma-derived sEVs, and DEMs were screened out between the above three groups. Regarding the distribution of DEMs in the NM-NSCLC and M-NSCLC groups, KEGG pathway analysis showed enrichment in focal adhesion and gap junctions and ClueGO in the Rap1 and Hippo signaling pathways; miEAA found that fibroblasts were over-represented. From our screening, miRNA-200c-3p and miRNA-4429 were found to be predictive DEMs among the CT, NM-NSCLC and M-NSCLC groups, and qRT‒PCR was applied to verify the results. Finally, it was revealed that expression levels of miR-200c-3p and miR-4429 were significantly upregulated in M-NSCLC patients.CONCLUSION:This study identified miRNA-200c-3p and miRNA-4429 as potential biomarkers for NSCLC metastasis.

ObjectiveCurrently, immune checkpoint inhibitors (ICIs) therapy is one of the main methods of treatment in non-small cell lung cancer (NSCLC). This study aimed to explore the risk factors of VTE and evaluate the effect of ICIs on VTE in patients with NSCLC.Research design and methodsWe retrospectively studied patients with NSCLC who were divided into VTE group and without VTE (Non-VTE) group. We identified the risk factors of VTE in NSCLC patients and evaluated the effect of ICIs on VTE in NSCLC patients.ResultsWe found that clinical stage III-IV (P = 0.015) and Khorana score (KS) >= 2 (P = 0.047) were independent risk factors for the occurrence of VTE in NSCLC, and treatment with ICIs reduced the risk of VTE occurrence (P = 0.028). There were no differences of survival rates in the 12-month (P = 0.449), 24-month (P = 0.412), or 36-month (P = 0.315) between the VTE and non-VTE groups. History of anti-angiogenic therapy (P = 0.033) and chronic obstructive pulmonary disease (COPD) (P = 0.046) were independent risk factors for VTE in NSCLC patients who were treated with ICIs.ConclusionThis study suggests that we should strengthen anticoagulant therapy when using ICIs for NSCLC patients with a history of anti-angiogenic therapy and COPD.

Lung cancer is the leading cause of cancer-related death worldwide. MicroRNAs (miRNAs) in circulating small extracellular vesicles (sEVs) have been suggested to be potential biomarkers for cancer diagnosis. The present study was designed to explore whether plasma-derived sEV miRNAs could be utilized as diagnostic biomarkers for differentiating between early-stage small cell lung cancer (SCLC) and early-stage non-small cell lung cancer (NSCLC). We compared the miRNA profiles of plasma-derived sEVs from healthy individuals, patients with early-stage SCLC and patients with early-stage NSCLC. Next-generation sequencing was used to screen for differentially expressed miRNAs (DEMs). Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were used to predict the potential functions of these DEMs. Weighted gene coexpression network analysis (WGCNA) was used to identify the different pathology-related miRNA modules. We found that 22 DEMs were significantly different among healthy individuals, patients with early-stage SCLC, and patients with early-stage NSCLC. We selected six representative DEMs for validation by qRT‒PCR, which confirmed that miRNA-483-3p derived from plasma sEVs could be used as a potential biomarker for the diagnosis of early-stage SCLC, miRNA-152-3p and miRNA-1277-5p could be used for the diagnosis of early-stage NSCLC respectively.

BACKGROUND:This study aimed to explore associations between PDL1 polymorphisms and efficacy of apatinib for patients with previously treated advanced non-small cell lung cancer (NSCLC) in a real-world setting.METHODS:We retrospectively recruited 148 patients with previously treated advanced NSCLC from January 2015 to December 2019 continuously. Clinical efficacy in patients receiving apatinib treatment was evaluated. Adverse reactions were documented during treatment. Biological specimens of peripheral blood and cancer tissue biopsies were obtained for the genotyping of genetic variations in PDL1 and corresponding gene-mRNA expression, respectively. Univariate association analysis between the status of PDL1 genetic variations and survival was performed with Kaplan-Meier survival analysis.RESULTS:The objective response rate of the 148 patients was 17.6% and disease-control rate 68.9%. Prognostic data suggested that median progression-free survival (PFS) was 3.8 (95% CI 3.13-4.47) months and median overall survival (OS) 10.5 (95% CI 9.06-11.95) months. Regarding PDL1 genetic variation, only rs2297136 was of clinical significance. Prognosis analysis revealed that PFS and OS for the rs2297136 genotype were significantly different. Median PFS of patients with TC/CC and TT genotypes was 3 and 4.5 months, respectively (P=0.006). Median OS of the two genotypes was 9 and 11.6 months, respectively (P=0.04). Furthermore, the safety profile suggested that the most common adverse reactions were hypertension, dermal toxicity, fatigue, and oral toxicity. This study failed to find any significant association between adverse reactions and rs2297136. Interestingly, mRNA-expression analysis demonstrated that mRNA expression of PDL1 in biopsy cancer-tissue specimens was significantly different based on rs2297136-genotype status (P<0.001).CONCLUSION:The PDL1 polymorphism rs2297136 could be used as a potential biomarker for the prognosis of patients with NSCLC receiving apatinib monotherapy.

Platelet-derived growth factor (PDGF) and its receptors (PDGFR), including PDGFR alpha and PDGFR beta, play important roles in tumorigenesis, tumor progression, and the regulation of stromal cell function. Constitutive activation of PDGFR signaling, gene rearrangement, and activating mutations of PDGFR have been identified in various types of human tumors and malignancies. PDGFR alpha and PDGFR beta belong to the family of type III receptor tyrosine kinases and, upon stimulation, activate downstream signaling cascades. Crenolanib is a specific tyrosine kinase inhibitor that targets and inhibits the kinase activity of PDGFR and the FMS-related tyrosine kinase 3. Its clinical efficacy in several human tumors is currently under investigation in Phase II clinical trials. In this study, we examined the potential role of crenolanib in the treatment of non-small-cell lung cancer (NSCLC). Using A549 cells as a model system, we have shown that crenolanib is capable of suppressing proliferation and inducing apoptosis in a dose-dependent manner. Crenolanib-treated cells have reduced migratory activity in response to inducers of chemotaxis. Furthermore, the in vivo antitumor activity of crenolanib was confirmed in an NSCLC xenograft tumor model. Injection of crenolanib significantly inhibited the growth of tumor mass by inducing apoptosis in tumor cells. Our results provide strong evidence supporting the use of crenolanib as a potential therapeutic agent in treating NSCLC. This work sets a foundation for further development of targeted and personalized therapeutics for lung cancer.

OBJECTIVE:To study the expression of hyperplasic suppress gene(HSG),epidermal growth factor receptor(EGFR) and their correlation in breast cancer tissues.METHODS:The method of real-time Fluorescene quantitative polymerase chain reaction(RT-PCR) were applied to detect the proliferation of HSG.EGFR mRNA and the protein expression of HSG,EGFR were detected by using SP immunohistochemistry in normal breast tissue,breast carcinoma,breast fibroadenoma tissues and their relevance in breast cancer tissues.RESULTS:The expression of HSG mRNA was significantly lower in breast cancer than those in breast fibroadenoma and normal breast tissue(P0.05);The expression of EGFR mRNA was significantly higher in breast cancer than those in breast fibroadenoma and normal breast tissue(P0.05);The expression of HSG mRNA and EGFR mRNA in breast fibroadenoma and normal breast tissues showed no statistically significant difference(P0.05).The expression of HSG and EGFR mRNA was correlated with lymph node metastasis,different pathological grades of breast cancer,ER and HER-2 and that was incorrelated with the age of the patient,tumor type,tumor size and clinical stage(P0.05).The expressions of HSG and EGFR were negative correlation in breast cancer tissues(r=-0.456,P=0.002).CONCLUSIONS:With the decline in the degree of differentiation or lymph node metastasis and the negative expression of ER and HER-2,the expression of HSG mRNA decreased significantly in breast cancer tissues.The expression level of EGFR is significantly increased in low pathological grades and the positive expression of ER and HER-2 of breast cancer.The expression of HSG is negatively correlated with EGFR in breast cancer tissues.The expression of HSG is positively correlated with inhibitting the occurrence of cancer tissue and the metastasis of lymph node.

Objective: To establish an ovarian cancer cells with high expression of LPA 1 by transfecting LPA 1 genes into the ovarian cancer cell lines and to study the biological characteristics of the transfected cells and to study the effects of lysophosphatidic acid (LPA) on the expressions and activities of MMP-2 and MMP-9 in ovarian cancer cells with stable expression LPA 1 genes. Methods: The pcDNA3. 1-LPA 1 was transfected into ovarian cancer cells line (SKOV3 and 3AO) with lipofectamine reagent. Control and vector groups were set up. Transfection and expression were confirmed by the means of RT-PCR. The biological characteristics of the transfected cells with expression LPA receptor genes were studied by the observation of their growth and proliferative period. The effects of LPA on the of MMP-2 and MMP-9 protein expressions and activities in transfected ovarian cells were analyzed by gelatin zymography. Results: It was confirmed that the LPA 1 genes had been stably integrated into 3AO and SKOV3 cell lines. The growth characteristics of transfected cells were not different before and after transfectation. The high expression of LPA 1 gene was revealed by the means of RT-PCR. The expression levels of LPA 1 mRNA in transfected cells (SKOV3-LPA 1 and 3AO-LPA 1) were significantly higher than those in control transfected cells and an empty vector groups, there were significant differences (P<0.05). The activities and protein expressions of MMPs in the cells transfected with LPA 1 receptor genes did not increase, compared with control and empty vector groups (P>0.05). Conclusion: The ovarian cancer cells with high expression of LPA 1 gene in vitro have been established. LPA 1 plays a role as negative growth receptor in the process of LPA-induced MMPs invasion and metastasis of ovarian cancer cells.

Accumulation of single nucleotide polymorphisms (SNPs) in the displacement loop (D-loop) of mitochondrial DNA (mtDNA) may be associated with an increased cancer risk. We investigated the lung cancer risk profile of D-loop SNPs in a case-controlled study. The minor alleles of nucleotides 235A/G and 324A/G were associated with an increased risk for lung cancer patients. The minor alleles of the nucleotides 151C/T, 200A/G, 524C/CA, and 16274G/A were specifically associated with the cancer risk of squamous cell carcinoma, whereas the minor allele of nucleotide 16298T/C was specifically associated with the risk of small cell lung cancer. In conclusion, SNPs in mtDNA are potential modifiers of lung cancer risk. The analysis of genetic polymorphisms in the mitochondrial D-loop can help identify subgroups of patients who are at a high risk of developing lung cancer.

Accumulation of single-nucleotide polymorphisms (SNPs) in the displacement loop (D-loop) of mitochondrial DNA (mtDNA) may be associated with disease outcome. Our team investigated the prediction power of D-loop SNPs in non-small cell lung cancer (NSCLC) outcome. In an overall multivariate analysis, allele 16390 was identified as an independent predictor for NSCLC outcome. The length of survival of patients with allele 16390A was significantly shorter than that of patients with allele 16390G (relative risk, 0.323; 95% CI, 0.109-0.951; p=0.040). The analysis of genetic polymorphisms in the mitochondrial D-loop can help identify NSCLC patient subgroups at a high risk for a poor disease outcome.

OBJECTIVE:To evaluate the influence of LPA1,LPA2 and LPA3 antisense oligodeoxynucleotides(ASODN) on inhibition of human ovarian cancer 3AO and SKOV3 cells,and study the effects of lysophosphatidic acid (LPA) on MMP-2 and MMP-9 expressions and activities in ovarian cancer cells with transfected LPA receptors ASODN. METHODS:The LPA1,LPA2 and LPA3 antisense oligodeoxynucleotides (ASODN) were integrated into 3AO and SKOV3 cells lines. Transfection and expression were confirmed by the means of RT-PCR and Western blotting. The biological characteristics of the transfected cells were studied by the observation of their growth and proliferative periods. The effect of LPA on MMP-2 and MMP-9 protein expressions and activities in transfected ovarian cells was analyzed by gelatin zymography. RESULTS:The LPA1 gene,LPA2 and LPA3 protein expressions were decreased in ovarian cancer cells with LPA1,LPA2 and LPA3 ASODN transfection. The transfected cells growth characteristics were not different before and after the transfectation. The gelatin zymography confirmed that LPA-induced the activities and protein expressions of MMP-2 and MMP-9 decreased in the transfected cells with LPA2 and LPA3 receptors ASODN,compared with those in the control group and SODN group (P0.05). The activities and protein expressions of MMPs in LPA1 receptors ASODN cells did not decrease,compared with those in the control group and SODN group (P0.05). CONCLSUIONS:LPA receptors antisense oligodeoxynucleotides can inhibite the expressions of LPA receptors gene and protein. LPA2 and LPA3 were more efficient in LPA-induced MMPs activity. LPA2 and LPA3 play important roles in the process of LPA-induced MMP invasion and metastasis of ovarian carcer cells.

OBJECTIVE:To investigate the lysophosphatidic acid (LPA) receptors expression situation and their biological significance in human ovarian cancer cell lines and in human epithelial ovarian neoplasms. METHODS:The reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were employed to measure the expression levels of LPA(1), LPA(2) and LPA(3) mRNA, LPA(2) and LPA(3) protein expression in cultured human ovarian cancer cell lines (3AO, SKOV3 and OVCAR3) and in human epithelial ovarian neoplasms. The immunocytochemical method was used to detect LPA(2) and LPA(3) protein expression in cultured human ovarian cancer cell lines. RESULTS:RT-PCR revealed that all ovarian cancer cell lines expressed LPA(1), LPA(2) and LPA(3) mRNA. The positive rates (100%; 86.4%; 88.2%) of LPA(1) mRNA in normal ovarian tissue, benign tumor and ovarian cancer were no significant difference (p>0.05). The expression level of LPA(1) mRNA was significantly higher in normal ovarian tissue compared with that in benign tumor and in ovarian cancer tissue (p<0.01). LPA(1) expression level was no significant difference in both benign tumor and ovarian cancer tissue (p>0.05). LPA(2) mRNA-positive rates (92.6%) and expression level were significantly higher in ovarian cancer compared with that in benign tumor (31.8%) and in normal ovarian tissue (31.3%) (p<0.01); LPA(2) mRNA-positive rates and expression level were no significant difference in both benign tumor and normal ovarian tissue (p>0.05). LPA(3) mRNA-positive rates (92.6%) and expression level were significantly higher in ovarian cancer compared with that in benign tumor (31.8%) and in normal ovarian tissue (31.3%) (p<0.01), LPA(3) mRNA-positive rates and expression level were no significant difference in both benign tumor and normal ovarian tissue (p>0.05). LPA(1) mRNA expression level was significantly decreased compared with that of LPA(2) and LPA(3) in ovarian cancer (p<0.01); Western blotting clearly revealed that all ovarian cancer cell lines showed LPA(2) and LPA(3) protein. The positive rates and expression level of LPA(2) and LPA(3) protein were significantly increased in ovarian cancer (92.6%; 92.6%) compared with that in benign tumor (45.5%; 45.5%) and that in normal ovarian tissue (43.8%; 43.8%) (p<0.01); LPA(2) and LPA(3) protein-positive rates and expression level were no significant difference in both benign tumor and normal ovarian tissue (p>0.05). Correlation of clinicopathological parameters showed that LPA receptors mRNA and protein expression were associated with FIGO stage and histological grade, except pathologic types and age. The mRNA and protein expression of LPA(2) and LPA(3) in stages III and IV was significantly higher than that in stages I and II epithelial ovarian cancer (p<0.05). The mRNA and protein expression of pathologic grade G(3) was significantly higher compared with grade G(1) (p<0.05). CONCLUSION:LPA(1), LPA(2) and LPA(3) mRNA and protein expressed widely in human epithelial ovarian neoplasms. LPA(2) and LPA(3) may be involved in the development and progression of human ovarian cancer. There was a significant correlation between LPA(2), LPA(3) and invasion and metastasis of epithelial ovarian cancer. LPA(2) and LPA(3) may be a prognostic indicator in patients with epithelial ovarian cancer.

OBJECTIVE:To investigate the expressions of lysophosphatidic acid (LPA) receptors at both mRNA and protein levels and their biological significance in different human ovarian cancer cell lines.METHODS:Reverse transcription-polymerase chain reaction (RT-PCR), streptovidin peroxidase-conjugated (SP) immunohistochemical assay and Western blotting were employed to measure the expression levels of LPA(1), LPA(2) and LPA(3) mRNA and LPA(2) and LPA(3) protein in cultured human ovarian cancer cell lines 3AO.SKOV3, and OVCAR3.RESULTS:All the ovarian cancer cell lines expressed LPA(1), LPA(2) and LPA(3) mRNAs and LPA(2) and LPA(3) proteins.CONCLUSION:LPA(1), LPA(2) and LPA(3) expressions occur extensively in human epithelial ovarian cancer cell lines, and LPA(2) and LPA(3) may be involved in the development and progression of human ovarian cancer.