Expression In Lung Cancer Tissues Research Articles

Correlation between DCE-MRI quantitative perfusion histogram parameters, apparent diffusion coefficient and Ki-67 in different pathological types of lung cancer

Objective: To explore the efficacy of dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) histogram molecular imaging index, apparent diffusion coefficient (ADC) in different types of lung cancer and explore their correlation with Ki-67. Methods: A total of 33 cases of lung cancer patients confirmed by pathology in Shaoxing People's Hospital from March 2017 to March 2018 were collected, 28 males and 5 females aged 50-85 years old, including 15 cases of squamous cell carcinoma, 12 cases of adenocarcinoma, and 6 cases of small cell carcinoma. All patients performed DCE-MRI and DWI imaging within one week before surgery or puncture. ADC values, DCE-MRI quantitative perfusion parameters by histogram metrics analysis (mean value, skewness, kurtosis, uniformity, entropy, energy, quantile) of K(trans), K(ep), V(e), and V(p) were then collected. Ki-67 expression in lung cancer tissue was detected by immunohistochemical method. One-way analysis of variance and least significant difference were used to compare the differences among the parameters of the three groups which were normal distribution and equal variances, while Kruskal-Walls test and Mann-Whitney U test were used to compare the parameters that did not conform to normal distribution or variance. Pearson correlation analysis was used to compare the correlations between quantitative perfusion histogram parameters, ADC values and immunohistochemical scores of Ki-67. Results: The Ki-67 count in small cell lung cancer(458±82, P=0.011) and squamous cell carcinoma(355±277, P=0.034)were significantly higher than that in adenocarcinoma (168±164). The correlation analysis showed that there was a significant negative correlation between ADC values and Ki-67 (P=0.018, r=-0.416). And V(e) (Q5, Q10) was negatively related to Ki-67 (P=0.017, r=-0.420; P=0.040, r=-0.366). In squamous cell carcinoma patients, V(e) (homogeneity) was significantly negatively correlated with the expression of Ki-67 (P=0.033, r=-0.570). K(trans)(homogeneity) and V(e) (homogeneity, Q5, Q10, Q25) were significantly positively correlated with ADC values (P value from 0.001 to 0.035, r value from 0.545 to 0.765). Conclusion: DCE-MRI quantitative perfusion histogram parameters, ADC value can evaluate the lung cancer cell proliferation activity in different pathological types.

Prognostic value of TIM-1 expression in human non-small-cell lung cancer

BackgroundT-cell immunoglobulin and mucin domain 1 (TIM-1) is an important co-stimulatory molecule which serves as a surface marker for T cell activation, especially for Th2 cells. Recently, many studies have also shown that TIM-1 can be abnormally expressed in human cancers and may have a potential role in promoting cancer progression.MethodsThe immunohistochemistry was used to examine the TIM-1 expression in human non-small-cell lung carcinoma (NSCLC) tissues. The cellular studies were performed to investigate the role of TIM-1 in the regulation of biological functions of human lung cancer cell lines.ResultsWe found that the TIM-1 expression was increased in human NSCLC tissues compared with the adjacent normal tissues, and the OS rate of NSCLC patients with higher TIM-1 expression was significantly lower compared with the ones with lower TIM-1 expression. The COX model showed that higher TIM-1 expression in lung cancer tissues could be used as an independent prognostic predictor for the patients. Furthermore, we depleted TIM-1 in NSCLC cell lines A549 and SK-MES-1, and the cellular functional studies also revealed that depletion of TIM-1 could significantly inhibit the cell viability as well as the abilities of migration and invasion. In addition, our microarray data showed that certain signaling pathways were altered and enriched after depletion of TIM-1. We subsequently verified that PI3K/Akt signaling pathway was involved in the TIM-1-mediated regulation of cellular functions in NSCLC cells.ConclusionOur findings supported the notion that TIM-1 could serve as a potential therapeutic target for NSCLC.

MicroRNA‑214 upregulates HIF‑1α and VEGF by targeting ING4 in lung cancer cells.

Previous reports have indicated a potential link between microRNA (miR)‑214 and hypoxia. In the present study, the biological functions and potential mechanisms of miR‑214 were determined, as well as its correlation with HIF‑1α signaling in non‑small cell lung cancer (NSCLC) cells. Quantitative polymerase chain reaction revealed that miR‑214 expression was upregulated in lung cancer tissues compared with adjacent normal tissues. miR‑214 mimics were transfected into A549 cells, and MTT, colony formation, invasion and wound healing assays were performed. It was demonstrated that miR‑214 mimic transfection promoted the invasion, proliferation and migration of A549 cells. Furthermore, miR‑214 inhibitor transfection decreased H1299 cell invasion, proliferation and migration. Next, the association between miR‑214 expression and the HIF‑1α signaling cascade was examined. It was demonstrated that miR‑214 mimics upregulated the expression of hypoxia‑inducible factor (HIF)‑1α, vascular endothelial growth factor (VEGF), adenylate kinase 3 and matrix metalloproteinase (MMP)2, whereas miR‑214 inhibitor downregulated the expression of these factors. Using prediction software, it was demonstrated that tumor suppressor ING4 was a target of miR‑214. A luciferase reporter assay confirmed that ING4 was a direct target of miR‑214. There was a negative correlation between ING4 and miR‑214 expression in lung cancer tissues. In addition, ING4 siRNA and plasmid was transfected into cells in order to validate its effect on HIF‑1α, MMP2 and VEGF expression. ING4 overexpression downregulated HIF‑1α and its targets MMP2 and VEGF, while ING4 siRNA upregulated HIF‑1α, MMP2 and VEGF. In conclusion, it was demonstrated that miR‑214 targeted ING4 in lung cancer cells, and upregulated the HIF‑1α cascade, leading to MMP2 and VEGF upregulation. This approach may help to clarify the role of miRNA in non‑small lung cancer and may be a new therapeutic target for non‑small lung cancer.

LncRNA MSTO2P promotes proliferation and autophagy of lung cancer cells by up-regulating EZH2 expression.

This study was designed to investigate the specific mechanism underlying the regulatory effect of long noncoding ribonucleic acids (lncRNAs) MSTO2P on lung cancer (LCa) cell proliferation and autophagy via regulating enhancer of zeste homolog (EZH2) expression. Quantitative real-time polymerase chain reaction (qRT-PCR) was applied to analyze the levels of MSTO2P and EZH2 in 40 pairs of LCa tissues and corresponding adjacent tissues, as well as in LCa cell lines (H1299, H23, A549) and human bronchial epithelial cells (BEAS-2B). Besides, the effect of MSTO2P on cell proliferation ability was detected by cell counting kit-8 (CCK-8) and plate cloning experiments. The interaction between MTS02P and EZH2 as well as their effects on cell autophagy ability were examined by qRT-PCR and Western blot. The qRT-PCR results showed that MSTO2P expression in LCa tissues was remarkably higher than that in adjacent tissues. Meanwhile, compared with human bronchial epithelial cells, the level of MSTO2P was remarkably up-regulated in LCa cells. After down-regulating MSTO2P, the cell proliferation ability was weakened, and the protein levels of autophagy-related genes including Agt5, LC-3I, and LC-3II were remarkably down-regulated. At the same time, EZH2 expression in LCa tissues was also remarkably up-regulated relative to adjacent tissues, and it was positively correlated with the expression of MSTO2P. In addition, after down-regulating MSTO2P, the EZH2 level was also remarkably reduced. Further experimental results revealed that EZH2 down-regulation could impair the cell proliferation ability and down-regulate the expressions of autophagy genes such as Agt5, LC-3I, and LC-3II. LncRNA MSTO2P promotes LCa cell proliferation and autophagy by up-regulating EZH2. Therefore, MSTO2P may be a potential therapeutic target for LCa.

Rho GTPase Activating Protein 24 (ARHGAP24) Silencing Promotes Lung Cancer Cell Migration and Invasion by Activating β-Catenin Signaling.

BackgroundRho GTPase activating protein (RhoGAPs) is an important negative regulator of the Rho signaling pathway that is involved in tumorigenesis in liver, colon, and renal cancer. However, the mechanism by which Rho GTPase activating protein 24 (ARHGAP24) regulates cell invasion and migration of lung cancer has not been fully explained.Material/MethodsIn this study, ARHGAP24 expression in lung cancer tissues and cell lines was measured by immunohistochemical and Western blot analysis. Transwell or wound healing analysis was performed to detect the cell migration and invasion of ARHGAP24 modulated A549 and NCI-H1975 cells with β-catenin inhibitor XAV-939 (10 μM) treatment, and the expression of MMP9, VEGF, and β-catenin protein was measured by Western blotting.ResultsOur results showed that ARHGAP24 expression was downregulated in lung cancer tissues and cell lines. pLVX-Puro-ARHGAP24 transfection in A549 cells significantly inhibited cell invasion and migration, along with increased E-cadherin and decreased MMP9, VEGF, Vimentin, and β-catenin protein expression. pLKO.1-ARHGAP24-shRNA transfection in NCI-H1975 cells significantly promoted cell invasion and migration, accompanied with decreased E-cadherin and increased MMP9, VEGF, and β-catenin protein expression. Moreover, NCI-H1975 cells with XAV-939 treatment showed decreased cell invasion and migration when compared with pLKO.1-ARHGAP24-shRNA transfection. ARHGAP24 silencing promoted the transcriptional activity of β-catenin in NCI-H1975 cells.ConclusionsOur findings indicate that ARHGAP24 silencing promotes lung cancer cell migration and invasion through activating β-catenin signaling.

Sulforaphane Inhibits the Acquisition of Tobacco Smoke-Induced Lung Cancer Stem Cell-Like Properties via the IL-6/ΔNp63α/Notch Axis.

Background: Tobacco smoke (TS) critically contributes to the development of lung cancer; however, the underlying molecular mechanisms remain unclear. The induction of cancer stem cells (CSCs) by TS represents an early event in tumor initiation. The lung cancer-related gene ΔNp63α is highly expressed in epithelial tissues and drives tumor formation and cancer stem cell properties. This study investigated the role of ΔNp63α in the long-term acquisition of TS-induced lung CSC-like properties.Methods: The expression levels of ΔNp63α, lung CSC markers, and interleukin (IL)-6 in lung carcinoma specimens were determined by western blotting and enzyme linked immunosorbent assays. Human bronchial epithelial (HBE) cells were chronically exposed to 2 % cigarette smoke extract for 55 passages, following which colony formation capacity, expression of proteins associated with malignant transformation, lung CSC markers, and tumor incidence were investigated. The effects of ΔNp63α on long-term TS exposure-induced lung CSC-like properties and Notch activation were analyzed using tumorsphere formation ability, immunofluorescence assays, luciferase reporter assays, and western blotting. The roles of IL-6 on chronic TS exposure-induced lung CSC-like properties and ΔNp63α expression were also examined. Moreover, the effects of sulforaphane (SFN) on TS-transformed lung CSC-like properties, IL-6 and ΔNp63α expression, and Notch signaling were investigated in vitro and in vivo.Results: Higher levels of ΔNp63α were observed in the lung cancer tissues of smokers than in those of non-smokers, whereas ΔNp63α was positively correlated with CD133 and Oct4 expression in lung cancer tissues. Data from the in vivo and in vitro experiments demonstrated that long-term TS exposure-transformed HBE (THBE) cells acquired lung CSC-like properties. Furthermore, ΔNp63α transcriptionally activated the Notch signaling pathway to promote the acquisition of CSC-like properties by the THBE cells. TS upregulated IL-6, which increased ΔNp63α expression in THBE sphere-forming cells. Finally, SFN inhibited the TS-induced CSC-like properties of THBE cells via the IL-6/ΔNp63α/Notch axis.Conclusion: Our data suggest that the IL-6/ΔNp63α/Notch axis plays an important role in the long-term TS exposure-induced acquisition of lung CSC-like properties and SFN intervention.

Overexpression of cancer-testis antigen melanoma-associated antigen A1 in lung cancer: A novel biomarker for prognosis, and a possible target for immunotherapy.

We investigated melanoma-associated antigen A1 (MAGE-A1) expression in lung cancer tissues and its correlation with prognostic factors. In this cross-sectional study, samples from 101 patients with lung cancer were obtained between 2007 and 2014 and stained for MAGE-A1 by immunohistochemistry. Correlation with prognostic factors was assessed by t test, and χ 2 , and Pearson's tests. Eighty non-small-cell lung cancer (NSCLC) and 21 small-cell lung cancer specimens were stained for MAGE-A1. MAGE-A1 was detected more commonly in adenocarcinomas and was expressed more frequently in male and patients >60 years. MAGE-A1 was found in several lung cancer patients. MAGE-A1 was expressed more commonly in NSCLC, elderly, and men. Further investigations are needed to assess MAGE-A1 as potential cancer biomarkers.

Secreted Phosphoprotein 1 Promotes the Development of Small Cell Lung Cancer Cells by Inhibiting Autophagy and Apoptosis.

This study aimed to investigate the expression of secreted phosphoprotein 1 (SPP1) on lung cancer cells and explore its underlying mechanism on autophagy and apoptosis which effect the development of lung cancer cells. GSE19804 related to lung cancer cells was screened from Gene Expression Omnibus (GEO) database, and we screened the 47 pairs of differential expressed mRNAs in lung cancer cells and adjacent tissues using microarray analysis. The expression of the core gene SPP1 was detected by qRT-PCR and western-blot. The transfection efficiency of lung cancer cells was detected by qRT-PCR and the expression of transfected group was tested by western-blot. Cell proliferation after transfection was tested by MTT assay and plate cloning experiment. The apoptosis rate of each transfection group was detected by flow cytometry. We use western-blot to test protein expression of autophagy-related proteins Beclin-1, LC3-I, LC3-II and p62 of each transfected group. Through analysis of GSE19804,the heat map showed SPP1 was the highest expressed in tumor tissues. qRT-PCR and western-blot detected SPP1 expression in lung cancer tissues was higher than that in normal adjacent tissues and was significantly increased in lung cancer cell lines. After transfection with pcDNA3.1-SPP1 (p-SPP1 group), siRNA1-SPP1 (siRNA1 group) and siRNA2-SPP1 (siRNA2 group), showed different expression of SPP1. Up-regulation of SPP1 enhanced cell viability and promoted tumor cell proliferation, while knockdown of SPP1 inhibited tumor cell proliferation. From the results of apoptosis rate, SPP1 inhibited the tumor cell apoptosis. However, in normal lung cell, SPP1 had no effect on cell proliferation and apoptosis. And to test autophagy-related proteins, we found that overexpression of SPP1 inhibited autophagy. High expression of SPP1 inhibited autophagy and apoptosis to promote the development of small cell lung cancer cells.

<em>FAM129B</em> promoted tumor invasion and proliferation via facilitating the phosphorylation of FAK signaling and associated with adverse clinical outcome of non-small cell lung cancer patients

BackgroundFamily with sequence similarity 129, member B (FAM129B), also called MINERVA, is upregulated and promotes tumor invasion in multiple types of cancer. However, the mechanism and clinicopathological significance of FAM129B remains unclear.Materials and methodsOnline KM-plotter tool and immunohistochemistry were used to predict the prognostic value of FAM129B expression in lung cancer tissues. Western blotting analysis, MTT, colony formation assay and matrigel invasion assay were performed after overexpressing or depleting FAM129B.ResultsIn this study, using the online KM-plotter tool, we found FAM129B was correlated with adverse outcome in non-small cell lung cancer (NSCLC) patients (P<0.001). Immunohistochemistry results revealed that FAM129B showed negative or dim expression in normal lung tissues while presented positive cytoplasmic expression in both squamous cell lung carcinoma and lung adenocarcinoma. The positive ratio of FAM129B in clinical NSCLC tissue samples (77/187, 41.2%) was significantly higher than that in normal lung tissue samples (8/68, 11.8%; P<0.001). FAM129B expression associated with advanced TNM staging (P<0.001) and positive regional lymph node metastasis (P<0.001). The results of Kaplan-Meier analysis suggested that the survival time of patients with positive FAM129B expression was significantly shorter than those with negatively FAM129B expression (P<0.001). Proliferation and invasion assay revealed that FAM129B prominently facilitated tumor proliferation and invasion in NSCLC cells. Western blotting results revealed that FAM129B upregulated the expression of MMP2 and Cyclin D1 by enhancing the phosphorylation of FAK at Tyr 397 and Tyr 925. Incorporation of FAK inhibitor in the medium significantly downregulated the phosphorylation of FAK and subsequently attenuated increasing expression of MMP2 and Cyclin D1 induced by FAM129B overexpression.ConclusionOur results indicated that FAM129B may be a new prognosis predictor of NSCLC patients and impact tumor invasion and proliferation of NSCLC cells through promoting the activation of FAK signaling.

KAP1 inhibits the Raf-MEK-ERK pathway to promote tumorigenesis in A549 lung cancer cells.

Aberrant activation of the Raf-MEK-ERK pathway has frequently been associated with various cancers, especially lung cancer. However, the key regulators of this pathway are largely unknown. Using functional proteomics screening, we found that KAP1 interacts with c-Raf. Knocking out KAP1 decreased c-Raf phosphorylation at serine 259 and increased its phosphorylation at serine 338, which activated MEK and ERK. We detected higher KAP1 expression in lung cancer tissues than in normal peri-tumoral tissues. KAP1 knockdown arrested A549 lung cancer cells in the G0/G1 phase of the cell cycle and attenuated cell growth, metastasis, the epithelial-mesenchymal transition, angiogenesis, stemness, and colony formation. Furthermore, knocking out KAP1 remarkably increased the susceptibility of A549 cells to the anti-cancer drug 5-Fluorouracil, which correlated with increasing ERK phosphorylation. In vivo xenograft experiments suggested that KAP1 deficiency significantly decreases the tumorigenicity of A549 cells. Taken together, our findings indicate that KAP1 acts as a key module in the c-Raf-interactome complex and regulates lung cancer development through the Raf-MEK-ERK pathway. Therefore, KAP1 may represent a potential diagnosis biomarker and new treatment target for lung cancer.

RAD52 motif‑containing protein1 promotes non‑small cell lung cancer cell proliferation and survival via cell cycle regulation.

DNA repair proteins such as RAD52 have been implicated in tumor progression and response to chemotherapy. RAD52 motif‑containing protein1 (RDM1) has been implicated in the response to chemotherapeutic agent cisplatin; however, its function in lung cancer progression remains unclear. This study aimed to investigate the role of RDM1 in the progression of non‑small cell lung cancer (NSCLC). We found elevated RDM1 mRNA and protein expression in NSCLC tissues and cell lines compared to levels in normal lung cells. RDM1 protein expression in lung cancer tissues was found to correlate with tumor size, histological differentiation, lymph node metastasis and tumor‑node‑metastasis (TNM) stage. Knockdown of the RDM1 gene with siRNA significantly reduced the cellular proliferation rate and increased apoptosis in the human NSCLC cell line, NCI‑H1299. Compared to wild‑type NCI‑H1299 cells, RDM1 knockdown enhanced the activity of caspase‑3 and caspase‑7, and decreased the proportion of cells in the S‑phase of the cell cycle. Taken together, these data imply that RDM1 promotes the survival and proliferation of NSCLC cells. Due to its similarity to RAD52, we hypothesized that RDM1 potentially repairs DNA double‑strand breaks arising through DNA replication, thereby preventing G2/M cell cycle arrest. Accordingly, specific targeting of RDM1 may be a novel therapeutic strategy in the treatment of NSCLC.

LncRNA NBAT-1 is down-regulated in lung cancer and influences cell proliferation, apoptosis and cell cycle.

To explore the expression and role of lncRNA NBAT-1 in lung cancer. LncRNA NBAT-1 expression in lung cancer tissues and adjacent areas was detected via reverse transcriptase-polymerase chain reaction (RT-PCR). RAC1 protein was analyzed via Western blotting assay. Cell counting kit-8 (CCK-8) and flow cytometry were used to evaluate cell proliferation and apoptosis as well as cell cycle. The expression level of lncRNA NBAT-1 in cancer specimen was remarkably lower than that in adjacent areas. Furthermore, the low expression of lncRNA NBAT-1 had a significant correlation with patient's tumor size, differentiation degree of tumor cells and lymph node metastasis. The overexpression of lncRNA NBAT-1 could inhibit the proliferation and cell cycle, promote the apoptosis of A549 cells, and down-regulate the expression level of RAC1. The low expression of lncRNA NBAT-1 is involved in the progression of lung cancer.

High CXC Chemokine Ligand 16 (CXCL16) Expression Promotes Proliferation and Metastasis of Lung Cancer via Regulating the NF-κB Pathway.

BackgroundsCXC chemokine ligand 16 (CXCL16) is a soluble chemokine with a transmembrane domain, playing an important role in inflammatory regulation. NF-κB has a critical role in tumor progression. Recent studies focused on the effect of CXCL16 on tumor progression. However, few reports showed the influence of CXCL16 on lung cancer, especially in regulating NF-κB activity. Here we investigated CXCL16 expression and its clinical significance in lung cancer, as well as the effect on lung cancer cell biological characteristics by regulating NF-κB.Material/MethodsCXCL16 expression in lung cancer was detected and its associations with clinical characteristics were analyzed. Proliferation and invasion of A549 and PC-9 cells was measured before and after silencing CXCL16 or inhibiting the NF-κB pathway, separately.ResultThe positive rate of CXCL16 in lung cancer tissue was significantly higher than that in adjacent tissue, and that in patients with lymphatic metastasis was significantly higher than that in patients without (all, P<0.05). The positive rate of CXCL16 was significantly (P<0.05) positively corrected with poor prognosis of lung cancer. Silencing CXCL16 not only suppressed proliferation and invasion of A549 and PC-9 cells, but also significantly (P<0.05) inhibited c-Rel, p105, and Rel-B in the NF-κB pathway. Inhibiting NF-κB also suppressed proliferation and invasion of A549 and PC-9 cells, which was similar to the results after silencing CXCL16.ConclusionsEnhanced CXCL16 expression in lung cancer tissue promoted the proliferation and invasion of lung cancer cells. CXCL16 might promote proliferation and invasion of lung cancer by regulating the NF-κB pathway.

TTF-1及TS在肺腺癌中的表达及与培美曲塞化疗疗效相关性分析

目的：探讨晚期肺腺癌中TTF-1和TS的表达与培美曲塞化疗疗效的关系。方法：回顾性分析80例接受培美曲塞化疗的晚期肺腺癌患者的临床病理资料。免疫组织化学法检测肿瘤标本TTF-1和TS蛋白的表达，并对疗效进行分析。结果：TTF-1阳性表达率为71.25% (57/80)，TS阳性表达率为22.50% (18/80)。TTF-1和TS在肺癌中的表达呈负相关性(r = −0.356，P = 0.002)。化疗有效率(CR + PR)为46.25%，TTF-1(+)患者有效率(64.91%)显著高于TTF-1(−)患者(39.13%)，差异有显著性(P < 0.05)。TS(+)患者有效率(27.78%)显著低于TS(−)患者51.61%，差异有显著性(P < 0.05)。联合检测显示：TTF-1(−)且TS(+)组有效率为28.57%，中位无疾病进展时间为3.9个月，1年生存率为28.6%，TTF-1(+)且TS(−)组有效率为81.13%，中位无疾病进展时间为7.4个月，1年生存率为69.8%，差异均有显著性(P < 0.05)。结论: 联合检测TTF-1和TS的表达可作为培美曲塞化疗方案治疗晚期肺腺癌的疗效预测指标之一。 Objective: To investigate the relationship between expression of TTF-1, TS and chemotherapy effect of pemetrexed in advanced lung adenocarcinoma. Methods: Eighty patients with advanced lung adenocarcinoma treating by pemetrexed were enrolled in this study and their tumor samples were collected retrospectively for analysis. The expression of TTF-1, TS protein in tumor samples was de-tected by the immunohistochemical method. The data of therapeutical effect and toxicity were col-lected and analyzed. Results: The TTF-1 positive staining accounted for 71.25% (57/80). The TS positive staining accounted for 22.50% (18/80). TTF-1 and TS expression in lung cancer tissues had negative correlation (r = −0.356, P = 0.002). The response rate (CR + PR) was 46.25%. The efficiency of patients with over-expression of TTF-1 (64.91%) was significant higher than patients with low-expression of TTF-1 (39.13%) (P < 0.05). The efficiency of patients with over-expression of TS (27.78%) was significant lower than patients with low-expression of TS (51.61%) (P < 0.05). The ef-ficiency, median progress time and 1-survival rate of patients with over-expression TS and low-expression of TTF-1 (28.57%, 3.9 months, 28.6%) were all significant lower than patients with over-expression TTF-1 and low-expression of TS (81.13%, 7.4 months, 69.8%) (P < 0.05). Conclu-sions: The expression of TTF-1 and TS is associated with chemotherapy effect of pemetrexed in ad-vanced lung adenocarcinoma and can be used to detect the sensitivity.

ZNF259 inhibits non-small cell lung cancer cells proliferation and invasion by FAK-AKT signaling.

BackgroundZinc finger protein 259 (ZNF259) is known to play essential roles in embryonic development and cell cycle regulation. However, its expression pattern and clinicopathological relevance remain unclear.Materials and methodsA total of 114 lung cancer specimens were collected. The ZNF259 expression was measured between the lung cancer tissues and the adjacent normal lung tissues by immunohistochemical staining and Western blotting. Moreover, the correlation of ZNF259 expression with clinicopathological features was analyzed in 114 cases of lung cancer. Additionally, ZNF259 was depleted in the lung cancer cells in order to analyze its effect in the lung cancer.ResultsImmunohistochemical staining of 114 lung cancer specimens revealed significantly lower ZNF259 expression in lung cancer tissues than in adjacent normal lung tissues (53.5% vs 71.4%, P<0.001). In addition, ZNF259 downregulation was significantly associated with larger tumor size (P=0.001), advanced TNM stage (P=0.002), and positive lymph node metastasis (P=0.02). Western blotting of 20 paired lung cancer samples revealed lower ZNF259 protein levels in lung cancer tissues than in those of corresponding normal lung tissues (P=0.0032). Depletion of ZNF259 resulted in enhanced levels of p-FAK and p-AKT, CyclinD1, and MMP2, which in turn increased the proliferation and invasion of lung cancer cells. The effects of ZNF259 depletion were reversed by treatment with specific FAK or AKT inhibitors.ConclusionZNF259 depletion is correlated with the development of non-small cell lung cancer (NSCLC) and serves as a predictor of adverse clinical outcome in NSCLC patients. The inhibitory effect of ZNF259 on proliferation and invasion can be attributed to downregulation of CyclinD1 and MMP2 via inactivation of the FAK-AKT pathway.

Associations between abnormal vitamin D metabolism pathway function and non-small cell lung cancer.

Lung cancer is a type of malignant tumor derived from the respiratory system, which is the leading cause of cancer-associated mortality worldwide, of which ~80% of cases are attributable to non-small cell lung cancer (NSCLC). A previous study demonstrated that 1α,25-Dihydroxyvitamin D3 (1α,25(OH)2D3), derived from the vitamin D metabolic pathway contributes an antitumor effect. Aberrant expression of the essential enzyme encoding genes, Cytochrome P450 Family 27 Subfamily A Member 1 (CYP27A1), Cytochrome P450 Family 27 Subfamily B Member 1 (CYP27B1), and Cytochrome P450 Family 24 Subfamily A Member 1 (CYP24A1) may be associated with lung cancer. However, a lack of evidence exists concerning the association between CYP27A1, CYP27B1, CYP24A1 expression and NSCLC. The aim of the present study was to investigate the functions of CYP27A1, CYP27B1 and CYP24A1 expression in NSCLC. Lung cancer tissue and para-carcinoma control tissue were collected from patients with NSCLC. Reverse transcription-quantitative polymerase chain reaction was applied to analyze CYP27A1, CYP27B1 and CYP24A1 mRNA expression in lung cancer tissues. An association analysis was performed between the aforementioned metabolic enzymes and patients with NSCLC age, gender, tumor node metastasis (TNM) stage, pathological type, differentiation and prognosis. CYP27B1 and CYP24A1 mRNA were upregulated in NSCLC compared with controls (P<0.05). However, no significant differences in CYP27A1 expression were observed between NSCLC and control. In addition, CYP24A1 expression was not associated with age, sex, smoking or TNM stage, but was associated with pathological type, differentiation and prognosis (P<0.05). CYP27B1 expression was significantly associated with TNM stage, differentiation, and prognosis, but not age, sex, smoking or pathological type. In conclusion, CYP27B1 and CYP24A1 may be considered as independent prognostic factors of NSCLC and may be novel therapeutic targets to assist clinical diagnosis, treatment and prognosis of the disease.

TMEM17 depresses invasion and metastasis in lung cancer cells via ERK signaling pathway.

Transmembrane protein 17(TMEM17) is a newly identified protein, its expression pattern and clinicopathological relevance is still unclear. In this study, western blot assay was performed in 20 paired lung cancer samples and found that TMEM17 protein levels were lower in lung cancer tissues than that in the corresponding normal lung tissues (p=0.010). Immunohistochemistry staining in 143 cases lung cancer specimens also showed that TMEM17 expression in lung cancer tissues were significantly lower than adjacent normal lung tissues (35.7% vs 63.2%, p<0.001). And negative TMEM17 expression was significantly associated with poor histological differentiation (p=0.027), advanced TNM stages (p=0.006), positive lymph node metastasis (p=0.002) and poor prognosis (p=0.002). After overexpressing TMEM17, levels of p-ERK and its downstream molecules, p-P90RSK and Snail, were down-regulated, while levels of Occludin and Zo-1 were up-regulated, which result in the inhibition of invasion and migration ability of lung cancer cells. The effects were reversed by the incorporation of specific ERK inhibitor PD98059. In conclusion, loss of TMEM17 correlates with the development of non-small cell lung cancer (NSCLC) and predicts adverse clinical outcome of NSCLC patients. The effect of TMEM17 on inhibiting invasion and migration may attribute to restoring Occludin and Zo-1 expression through inactivating ERK-P90RSK-Snail pathway.

HMGA2 regulates lung cancer proliferation and metastasis.

BackgroundThis study aimed to explore the effects of HMGA2 on cell proliferation and metastases in lung cancer and its underlying mechanism.Methods HMGA2 expression in lung cancer tissues and its association with overall survival were analyzed based on data from a public database. The roles of HMGA2 were validated via loss‐of‐function and gain‐of‐function experiments in vitro. HMGA2 regulation by microRNA‐195 (miR‐195) was validated by real time‐PCR, Western blotting, and luciferase reporter assays.Results HMGA2 was upregulated and associated with reduced overall survival in patients with lung adenocarcinoma. HMGA2 knockdown suppressed the proliferation and motility of H1299 cells, while HMGA2 ectopic expression in A549 cells increased cell proliferation and migration. HMGA2 affected cell apoptosis through caspase 3/9 and Bcl‐2, and regulated epithelial‐to‐mesenchymal transition by targeting Twist 1. Moreover, miR‐195 was found to directly target the 3′ untranslated region of HMGA2 messenger RNA and suppress its expression in lung cancer.ConclusionThis study demonstrated that HMGA2, regulated by miR‐195, played important roles in proliferation, metastases, and epithelial‐to‐mesenchymal transition in lung cancer. HMGA2 might serve as a biomarker and potential therapeutic target for lung cancer treatment.

Alex3 suppresses non-small cell lung cancer invasion via AKT/Slug/E-cadherin pathway.

Alex3, is a newly identified mitochondrial protein, regulates mitochondrial dynamics and is involved in neural development. However, its expression pattern and clinicopathological relevance in human tumors are still unclear. In this study, Immunohistochemistry assay was performed in 109 cases of lung cancer samples and found that Alex 3 expression in lung cancer tissues was significantly lower than adjacent normal lung tissues (28.4% vs 52.6%, p < 0.001). Sequent statistical analysis indicated that negative Alex3 expression was significantly associated with advanced tumor-node-metastasis stages (p = 0.001), positive lymph node metastasis (p = 0.005), and poor prognosis (p = 0.008). After overexpression of Alex3, levels of p-AKT and Slug were downregulated, while level of E-cadherin was upregulated, which results in the inhibition of invasion and migration ability of lung cancer cells. In conclusion, reduction of Alex3 correlates with the development of non-small cell lung cancer and predicts adverse clinical outcome of non-small cell lung cancer patients. The effect of Alex3 on inhibiting invasion and migration may attribute to upregulation of E-cadherin expression through AKT-Slug pathway inactivation.

MiR-26a enhances invasive capacity by suppressing GSK3β in human lung cancer cells

Lung cancer is the common cause of death from cancer, and most lung cancer patients die of metastasis. MicroRNAs (miRNAs) function as either oncogenes or tumor suppressors, playing crucial role not only in tumorigenesis, but also in tumor invasion and metastasis. There are several studies showed that miR-26a is involved in carcinogenesis, however, its role in tumor metastasis need to be elucidated. In this study, we showed that ectopic expression of miR-26a enhanced migration and invasion of lung cancer cells. Glycogen synthase kinase-3β (GSK3β) was identified as a direct target of miR-26a. GSK3β expression negatively correlated with miR-26a expression in lung cancer tissues. Silencing of GSK3β achieved similar effect as miR-26a over-expression; over-expression of GSK3β reversed the enhanced effect of miR-26a on lung cancer cell migration and invasion. Further study indicated that miR-26a increased β-catenin expression and nuclear translocation. C-myc and cyclin D1, the downstream genes of β-catenin, were also up-regulated by miR-26a. Furthermore, xenograft study showed that miR-26a promoted lung cancer cell growth in vivo, and suppressed GSK3β expression. Collectively, our results demonstrated that miR-26a enhanced metastatic potential of lung cancer cells via activation of β-catenin pathway by targeting GSK3β, suggesting the potential applicability of miR-26a as a target for cancer treatment.