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Background: Lung adenocarcinoma (LUAD) is the most common subtype of lung malignancy. However, the expression of cell division cycle-associated protein-3 (CDCA3) and its significance in LUAD remain unclear. In this study, we investigated the functional role of CDCA3 in LUAD through bioinformatics analysis and expected to provide a new direction for clinical treatment. Methods: The expression of CDCA3 was analyzed by online database. The association between the expression of CDCA3 and clinical parameters with LUAD was explored in TCGA. Survival and independent prognostic analysis were performed by TCGA database and the GSE30219 and GSE31210 datasets. Furthermore, Enrichment analyses were conducted to analyze the functions of CDCA3. Afterward, the relationship between CDCA3 and immune infiltration was investigated. Additionally, a competing endogenous RNA (ceRNA) regulatory network related to CDCA3 was constructed. Finally, CDCA3 expression was validated in clinical tissues by immunohistochemistry (IHC), real-time quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), and western blotting (WB). Results: CDCA3 expression was upregulated in 20 tumors and was significantly higher in LUAD compared with normal tissues in3 datasets. In addition, CDCA3 was significantly correlated with age, gender, stage, N, and smoking status. Kaplan-Meier survival curves showed that LUAD samples with higher CDCA3 expression were associated with poorer overall survival (OS) and disease-free survival (DFS). Univariate Cox regression analysis showed that the p value of CDCA3 expression was less than 0.05 (P<0.05) and it appeared in the results of multivariate Cox regression analysis (HR ≥1), indicating that CDCA3 can be used as an independent prognostic factor for LUAD. Intriguingly, Gene Set Enrichment Analysis (GSEA) suggested that CDCA3 was correlated with DNA-related terms and metabolic-related pathways in LUAD. CDCA3 expression was correlated with four immune scores and 14 immune cells in different groups. Next, a ceRNA network was constructed with CDCA3, and the experimental results of IHC, qRT-PCR, and WB were consistent with the bioinformatic analysis. Conclusions: CDCA3 could serve as a prognostic biomarker for LUAD.
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BACKGROUND: The cellular immunity of lung cancer patients is mainly the immune response of T cells, which plays an important role in tumour cell killing and immune surveillance. Transforming growth factor 1 (TGF-ß1) is secreted by tumour cells that can suppress the immune response and is an important group of immune down-regulation factors. Our study aims to investigate the effect of TGF-ß1 on the morphology and cellular immune function of A549 and peripheral blood mononuclear cells (PBMCs). METHODS: A549 cell line was cultured, PBMCs were cultured with different concentrations of TGF-ß1, and the morphology of A549 cells and PBMCs were seen. The levels of interleukin (IL)-2, IL-4, IL-6, IL-10, IFN-γ, and TNF and the numbers of CD3, CD4, CD8, CD4/CD8, and CD3 CD25 and CD4 CD25 in PBMCs were detected. RESULTS: During co-culture of A549 with PBMCs, TGF-ß1 can induced A549 showing epithelial-to-mesenchymal transition, enhanced its ability of migration and infiltration. Simultaneously, TGF-ß1 can depressing the growth and proliferation of PBMCs, inhibiting T-cell activation, and accelerating the PBMCs apoptosis. TGF-ß1 can inhibits A549 Th1 related-cytokines, enhance Th2 related-cytokines, cause the disorder of Th1/Th2, resulting in the Th1 cellular dominate immunity decline. CONCLUSIONS: TGF-ß1 may affect the secretion of related cytokines, hinder the activation of T lymphocytes, destroy the immune surveillance and killing effect of the body, and thus inhibit the cellular immunity.
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BACKGROUND: The role of miRNAs in non-small cell lung cancer (NSCLC) has been broadly studied and confirmed, and miR-107 has attracted an ever-growing level of attention. This study set out to research the mechanism of the effect of miR-107 on the malignant biological behavior of NSCLC in vivo and vitro. METHODS: The expression of miRNAs related to the development of NSCLC was detected by RT-qPCR. Western blotting was carried out to detect expression levels of serine/threonine kinase 33 (STK33) and proteins related to the extracellular regulated protein kinases (ERK) signaling pathway, while cell proliferation was detected using cell counting kit-8 (CCK-8). The cell apoptosis rate was measured using flow cytometry. The invasion ability was detected by Transwell assay. In vivo tumor growth assays were performed on mice. The expression ERK signaling pathway-related proteins in vivo was evaluated by immunohistochemistry staining. The targeted relationship between miR-107 and STK33 was confirmed by the dual luciferase reporter gene. RESULTS: In NSCLC cell lines and tissues, miR-107 was downregulated. Overexpression of miR-107 inhibited malignant biological behavior of NSCLC cell lines, and suppressed tumor growth in vivo. In addition, STK33 is one of the target genes of miR-107. Therefore, miR-107 suppressed cell proliferation and invasion and promoted tumor growth in vivo and cell apoptosis of NSCLC in vitro. The mechanism was found to be miR-107 targeting STK33, and a lack of STK33 led to the activation of ERK signaling pathway. CONCLUSIONS: miR-107 inhibited malignant biological behavior of NSCLC through regulation of the STK33/ERK signaling pathway.
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BACKGROUND: Lung cancer represents the most leading causes of cancer-related deaths worldwide, especially in Xuanwei in eastern Yunnan province, China. WD repeat and SOCS box containing protein (WSB) has been reported to participate in the carcinogenesis of lung cancer. However, there is no report about the role of WSB2 in the carcinogenesis and development of lung cancer in Xuanwei. Here, we investigated the functional role of WSB2 in Xuanwei lung cancer and uncovered its underlying molecular mechanisms. METHODS: The expression of WSB2 in lung cancer cell lines and tissues were measured using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Western blotting was used to determine the protein levels of WSB2, E-cadherin, N-cadherin, vimentin, c-Myc and ß-catenin in lung cancer cells. Cell viability was detected using 3-(4,5-diethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-etrazolium, inner salt (MTS) assay. While cell apoptosis and cell cycle distribution were quantified using flow cytometry following indicated staining. The change of cell invasion ability was detected using Transwell assay. FH535 was employed to block Wnt/ß-catenin pathway. A xenograft tumor model was applied to confirm the tumor properties of WSB2 in vivo. RESULTS: Our data showed that WSB2 was frequently up-regulated in Xuanwei lung cancer tissues and cells, when compared with paired non-cancerous tissues and normal lung epithelial cells. Knockdown of WSB2 notably reduced cell viability, cell invasion, epithelial-mesenchymal transition (EMT) process, while induced apoptotic cell death and cell cycle arrest of Xuanwei lung cancer cells. Moreover, in vivo findings also confirmed that WSB2 knockdown could effectively delay the growth of tumor. Mechanistic studies revealed that c-Myc and ß-catenin were notably decreased at both protein and mRNA levels after knocking down of WSB2, while overexpression of WSB2 showed a contrary tendency. In addition, blocking Wnt/ß-catenin pathway using FH535 rescued the cancer promoting effect mediated by overexpression of WSB2. Furthermore, WSB2 activated Wnt/ß-catenin pathway and accelerated the progression of lung cancer. CONCLUSIONS: WSB2 promoted the progression of lung cancer in Xuanwei by triggering Wnt/ß-catenin signaling pathway.
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A novel paradigm in tumor biology suggests that non-small cell lung cancer (NSCLC) growth is driven by lung cancer stem cell-like cells (LCSCs), but molecular mechanisms regulating tumorigenic and self-renewal potential of LCSCs are still unclear. Here, we aim to investigate biological function of SLC34A2 in regulating tumorigenicity of LCSCs and its underlying mechanisms. Our findings testified that CD166(+) cells which were derived from fresh primary NSCLC samples displayed stem cell-like features. Fluorescence-activated cell sorting (FACS) analysis showed the presence of a variable fraction of CD166 cells in 15 out of 15 NSCLC samples. Significantly, CD166(+) LCSCs from primary NSCLC tumors expressed high level of SLC34A2 which was required for CD166(+) LCSCs tumorigenic and self-renewal potential. In NSCLC patient cohort, increased SLC34A2 expression correlated with histology, which suggests a potential role of SLC34A2 in CD166(+) LCSCs. Furthermore, Wnt/ß-catenin pathway and Bmi1 were found necessary for tumorigenicity and self-renewal capacity of CD166(+) LCSCs by a series in vitro and in vivo experiments. Then, our study indicated that SLC34A2 regulated Bmi1 to promote tumorigenic and self-renewal potential of CD166(+) LCSCs through Wnt/ß-catenin pathway. In this study, the characterization of molecular basis of SLC34A2 in CD166(+) LCSCs not only allows for better understanding of the mechanisms regulating tumorigenicity of this specific population of NSCLC cells but also provides insight into the gradual improvement of more effective cancer therapies against this disease.
