转录因子Hey1在非小细胞肺癌中的表达及临床意义

目的检测Hey1与Notch1在非小细胞肺癌组织中的表达,探讨Hey1在非小细胞肺癌中的表达与Notch1的表达、各临床病理特征之间的关系。方法制作组织芯片,采用免疫组织化学染色检测Notch1和Hey1在246例非小细胞肺癌组织及相应117例癌旁正常肺组织中的表达,应用SPSS 21.0进行统计分析。结果 Notch1在肺鳞癌、肺腺癌和癌旁正常肺组织的阳性率分别为78.8%,28.9%和30.8%,Hey1在肺鳞癌、肺腺癌和癌旁正常肺组织中的阳性率分别为79.7%,23.4%和33.3%;肺鳞癌患者Notch1阳性表达与TNM分期和淋巴结转移情况呈正相关;肺鳞癌患者Hey1阳性表达与TNM分期呈正相关,与肿瘤分化程度呈负相关;Spearman相关分析显示,肺鳞癌和肺腺癌组织中Notch1表达与Hey1表达具有显著的正相关关系。结论 Hey1和Notch1与肺鳞癌的恶性程度密切相关,可能是肺鳞癌发生、发展的重要参与因子。     

ERCC-1和RRM-1在NSCLC中的表达及其病理临床意义

目的检测核苷酸切除修复(NER)系统的ERCC-1和RRM-1在非小细胞肺癌(NSCLC)组织中的表达并分析其表达的病理临床意义,为探索NSCLC的有关发生机制及术后辅助化疗提供有用的实验依据。方法应用免疫组织化学二步法检测128例NSCLC组织中ERCC-1和RRM-1的表达,以10例支气管扩张症肺组织作为对照。结果1.ERCC-1在NSCLC组织中的总体阳性表达率为44.53%(57/128),在对照组肺组织气道上皮的阳性表达率为100%(10/10),ERCC-1在NSCLC组织和对照组的阳性率差异有统计学意义(P<0.05);RRM-1在NSCLC组织中的总体阳性表达率为75%(96/128),在对照组肺组织气道上皮不表达,RRM-1在NSCLC组织和对照组的阳性率差异有统计学意义(P<0.05)。2.ERCC-1的表达与NSCLC患者的年龄、组织学类型及分化程度存在相关性(P<0.05);RRM-1的表达与NSCLC患者的年龄存在相关性(P<0.05)。NSCLC组织中ERCC-1与RRM-1的表达呈正相关(P<0.01)。结论ERCC-1表达降低或缺失可能与NSCLC发生有关;NSCLC组织中存在...     

MTA1-TJP1 interaction and its involvement in non-small cell lung cancer metastasis

Distant metastasis is the main cause of death in non-small cell lung cancer (NSCLC) patients. The mechanism of metastasis-associated protein 1(MTA1) in NSCLC has not been fully elucidated. This study aimed to reveal the mechanism of MTA1 in the invasion and metastasis of NSCLC.Bioinformatics analysis and our previous results showed that MTA1 was highly expressed in NSCLC tissues and correlated with tumor progression. Knockout of MTA1 by CRISPR/Cas9 significantly inhibited the migration and invasion of H1299 cells, but enhanced cell adhesion. Stable overexpression of MTA1 by lentivirus transfection had opposite effects on migration, invasion and adhesion of A549 cells. The results of in vivo experiments in nude mouse lung metastases model confirmed the promotion of MTA1 on invasion and migration. Tight junction protein 1 (TJP1) was identified by immunoprecipitation and mass spectrometry as an interacting protein of MTA1 involved in cell adhesion. MTA1 inhibited the expression level of TJP1 protein and weakened the tight junctions between cells. More importantly, the rescue assays confirmed that the regulation of MTA1 on cell adhesion, migration and invasion was partially attenuated by TJP1.In Conclusion, MTA1 inhibits the expression level of TJP1 protein co-localized in the cytoplasm and membrane of NSCLC cells, weakens the tight junctions between cells, and changes the adhesion, migration and invasion capabilities of cells, which may be the mechanism of MTA1 promoting the invasion and metastasis of NSCLC. Thus, targeting the MTA1-TJP1 axis may be a promising strategy for inhibiting NSCLC metastasis.     

Expression of monocyte chemoattractant protein-1 and CC chemokine receptor 2 in non-small cell lung cancer and its significance

Expression of monocyte chemoattractant protein-1 (MCP-1) and CC chemokine receptor 2 (CCR2) and its significance has been demonstrated in some cancer cells in recent clinical studies. However, the role of tumor MCP-1 and CCR2 expression in non-small cell lung cancer (NSCLC) remains unknown. The aim of the present study was to investigate the prognostic significance of MCP-1 and CCR2 expression in NSCLC cells. The relationship between MCP-1 and CCR2 expression in NSCLC cancer cells was examined by immunohistochemical staining of surgical specimens from 134 patients. Sixty-five of these patients had follow-up records. Kaplan–Meier analysis and Cox regression model were used to assess overall survival according to the presence or absence of MCP-1 and CCR2 expression in tumor cells. MCP-1 was detected in cancer cells of 107 NSCLC (79.9 %) and CCR2 was detected in cancer cells of 39 NSCLC (29.1 %). MCP-1 expression was correlated with sex, smoking habits, histology, and tumor size. Presence of MCP-1 in tumor cells was associated with better overall survival (P = 0.018). By multivariate analysis, MCP-1 expression in cancer cells showed an independent prognostic factor for overall survival (P = 0.002, hazard ratio [HR] = 0.256, 95 % confidence interval [CI] = 0.106–0.616). There was no significant relationship between CCR2 expression in tumor cells and clinical and pathological characteristics. Also, no significant positive correlation between MCP-1 and CCR2 expression was revealed by Spearman correlation analysis. Our data indicate that MCP-1 is overexpressed in NSCLC cells. Its expression in cancer cells is associated with better survival in NSCLC patients.     

ERCC1 expression as a prognostic and predictive factor in patients with non-small cell lung cancer: a meta-analysis

It is hypothesized that high expression of the excision repair cross-complementation group 1 (ERCC1) gene might be a positive prognostic factor, but predict decreased sensitivity to platinum-based chemotherapy. Results from the published data are inconsistent. To derive a more precise estimation of the relationship between ERCC1 and the prognosis and predictive response to chemotherapy of non-small cell lung cancer (NSCLC), a meta-analysis was performed. An electronic search of the PubMed and Embase database was performed. Hazard ratio (HR) for overall survival (OS) was pooled in early stage patients received surgery alone to analyze the prognosis of ERCC1 on NSCLC. HRs for OS in patients received surgery plus adjuvant chemotherapy and in patients received palliative chemotherapy and relative risk (RR) for overall response to chemotherapy were aggregated to analyze the prediction of ERCC1 on NSCLC. The pooled HR indicated that high ERCC1 levels were associated with longer survival in early stage patients received surgery alone (HR, 0.69; 95% confidence interval (CI), 0.58–0.83; P = 0.000). There was no difference in survival between high and low ERCC1 levels in patients received surgery plus adjuvant chemotherapy (HR, 1.41; 95% CI, 0.93–2.12; P = 0.106). However, high ERCC1 levels were associated with shorter survival and lower response to chemotherapy in advanced NSCLC patients received palliative chemotherapy (HR, 1.75; 95% CI, 1.39–2.22; P = 0.000; RR, 0.77; 95% CI, 0.64–0.93; P = 0.007; respectively). The meta-analysis indicated that high ERCC1 expression might be a favourable prognostic and a drug resistance predictive factor for NSCLC.     

Overexpression of polo-like kinase 1 and its clinical significance in human non-small cell lung cancer

Polo-like kinase 1 is a serine/threonine kinase which plays an essential role in mitosis and malignant transformation. The aim of this study was to investigate the prognostic significance of polo-like kinase 1 expression and determine its possibility as a therapeutic target in non-small cell lung cancer. Semi-quantitative RT-PCR assay was performed to detect polo-like kinase 1 mRNA expression in non-small cell lung cancer cells or tissues. Immunohistochemistry was performed to detect polo-like kinase 1 protein expression in 100 non-small cell lung cancer tissue samples, and the associations of polo-like kinase 1 expression with clinicopathological factors or prognosis of non-small cell lung cancer patients were evaluated. RNA interference was employed to inhibit endogenous polo-like kinase 1 expression and analyzed the effects of polo-like kinase 1 inhibition on the malignant phenotypes of non-small cell lung cancer cells including growth, apoptosis, radio- or chemoresistance. Also, the possible molecular mechanisms were also investigated. The levels of polo-like kinase 1 mRNA expression in non-small cell lung cancer cell lines or tissues were significantly higher than those in normal human bronchial epithelial cell line or corresponding non-tumor tissues. High polo-like kinase 1 expression was significantly correlated with advanced clinical stage, higher tumor classification and lymph node metastasis of non-small cell lung cancer patients (P = 0.001, 0.004 and 0.001, respectively). Meanwhile, high polo-like kinase 1 protein expression was also an independent prognostic molecular marker for non-small cell lung cancer patients (hazard ratio: 2.113; 95% confidence interval: 1.326-3.557; P = 0.017). Polo-like kinase 1 inhibition could significantly inhibit in vitro and in vivo proliferation, induce cell arrest of G₂/M phase and apoptosis enhancement in non-small cell lung cancer cells, which might be activation of the p53 pathway and the Cdc25C/cdc2/cyclin B1 feedback loop. Further, inhibition of polo-like kinase 1 could enhance the sensitivity of non-small cell lung cancer cells to taxanes or irradiation. Thus, polo-like kinase 1 might be a prognostic marker and a chemo- or radiotherapeutic target for non-small cell lung cancer.     

survivin和caspase-3在非小细胞肺癌中的表达及相关性研究

目的：研究survivin和caspase-3蛋白在非小细胞肺癌中的表达情况,分析二者的作用机制及相关性。方法：采用免疫组化KIT染色法,检测survivin和caspase-3在60例NSCLC组织和20例正常肺组织中的表达情况。结果：肺癌组织中的survivin阳性表达率（61．67％）明显高于正常肺组织（5％）;Ⅲ期的survivin阳性表达率（76．92％）明显高于Ⅰ＋Ⅱ期（50％）;均有显著性差异（P〈0．05）;而与年龄,病理类型,组织分化程度,淋巴结转移情况无关。肺癌组织中caspase-3阳性表达率（43．33％）明显低于正常组织（85％）;高＋中分化组的caspase-3阳性表达率（58．06％）明显高于低分化的（27．59％）;鳞癌组织中的caspase-3阳性表达率（56．25％）高于腺癌组织（28．57％）;均有显著性差异（P〈0．05）;而与年龄,TNM分期,淋巴结转移情况无关。结论：survivin在非小细胞肺癌中表达上调,而caspase-3却下降,且二者的表达呈负相关,提示survivin可能参与肺癌的发生发展,且抑制caspase-3的激活而抑制凋亡;caspase-3的失活可能导致肿瘤细胞的增殖。     

转录因子Snail在非小细胞肺癌中的表达及意义

目的检测转录因子Snail mRNA及蛋白在非小细胞肺癌(non-small cell lung cancer,NSCLC)中的表达,探讨Snail在NSCLC发生发展中的作用。方法采用免疫组织化学、RT-PCR的方法检测NSCLC、癌旁相对正常肺组织(>5cm)中转录因子Snail蛋白及mRNA的表达,并分析其与临床病理参数之间的关系。结果 NSCLC组织中Snail mRNA的阳性表达率(82.7%)高于癌旁相对正常肺组织(23.1%,P<0.05);NSCLC组织中Snail mRNA的阳性表达量(1.368±0.614)高于癌旁相对正常肺组织(0.579±0.217,P<0.05)。NSCLC组织中Snail蛋白的阳性表达率(80.8%)高于癌旁相对正常肺组织(23.1%,P<0.05),且癌组织中Snail蛋白阳性表达率与TNM分期、淋巴结转移有关(P<0.05),而与组织分型、分化程度、吸烟史无关(P>0.05)。结论 NSCLC的癌变、侵袭、转移可能与Snail蛋白、mRNA的高表达有关。     

Knockdown of LncRNAZFAS1 suppresses cell proliferation and metastasis in non-small cell lung cancer

ABSTRACT

To evaluate the effects of LncRNAZFAS1 on cell proliferation and tumor metastasis in non-small cell lung cancer (NSCLC), we detected the expression level of LncRNAZFAS1 in NSCLC-related tissues and cells. qRT-PCR results revealed that LncRNAZFAS1 in tumor tissues was significantly higher than that in normal lung tissue, especially significantly up-regulated in stage III / IV and in metastatic NSCLC tissues. LncRNAZFAS1 expression was dramatically up-regulated in 4 NSCLC-related cells (A549, SPC-A1, SK-MES-1, and NCI-H1299), with having the highest expression level in A549 cells. Furthermore, we implemented a knockdown of LncRNAZFAS1 in A549 cells, and the results of CCK8 and Transwell assays suggested that knockdown of LncRNAZFAS1 significantly inhibited NSCLC cell proliferation and metastasis. Next, we constructed a tumor xenograft model to evaluate the effect of LncRNAZFAS1 on the NSCLC cell proliferation in vivo. The results indicated that knockdown of LncRNAZFAS1 dramatically inhibited A549 cells proliferation and repressed tumor growth. Additionally, knockdown of LncRNAZFAS1 drastically weakened the expressions of MMP2, MMP9 and Bcl-2 proteins, whereas noticeably strengthened the expression of BAX protein. Our results altogether suggest that knockdown of LncRNAZFAS1 has a negative effect on the proliferation and metastasis of NSCLC cell, which implying LncRNAZFAS1 is a potential unfavorable biomarker in patients with NSCLC.     

榄香烯治疗非小细胞肺癌的作用机制研究进展

榄香烯注射液是从天然中草药姜科植物温郁金中提取获得的萜烯类化合物,其主要生物学活性为降低肿瘤细胞有丝分裂能力,诱导肿瘤细胞凋亡,抑制肿瘤细胞的生长,从而改善NSCLC患者的临床症状,增强患者免疫力.本文主要从杀灭癌细胞、诱导癌细胞凋亡、抑制癌细胞血管形成、增强患者免疫力等几方面综述榄香烯注射液对非小细胞肺癌的作用机制.     

Frequent epigenetics inactivation of KEAP1 gene in non-small cell lung cancer

The KEAP1/Nrf2 pathway is a master regulator of several redox-sensitive genes implicated in resistance of tumor cells against chemotherapeutic drugs. Recent data suggest that epigenetic mechanisms may play a pivotal role in the regulation of KEAP1 expression. We performed a comprehensive genetic and epigenetic analysis of the KEAP1 gene in 47 non-small cell lung cancer tissues and normal specimens. Promoter methylation analysis was performed using a quantitative methylation specific PCR assay in real time. Methylation at the KEAP1 promoter region was detected in 22 out of the 47 NSCLCs (47%) and in none of the normal tissues analyzed. Somatic mutations were detected in 7 out of the 47 tumors (15%) and loss of heterozygosity (LOH) in 10 out of the 47 (21%) of the cases. Overall, we found at least one molecular alteration in 57% of the cases. Approximately one third of the tumors had two alterations and this feature was associated with higher risk of disease progression in univariate COX regression analysis (HR = 3.62; 95% CI 1.24–10.65, p = 0.02). This result was confirmed by Kaplan-Meier analysis, which demonstrated an association between worst outcome and KEAP1 double alterations (p = 0.01, Log rank test). Our results further suggest that deregulation of the NRF2/KEAP1 system could play a pivotal role in the cancerogenesis of NSCLC. In addition identifying patients with KEAP1 genetic and epigenetic abnormalities may contribute to disease progression prediction and response to therapy in lung cancer patients.     

DJ-1 may contribute to metastasis of non-small cell lung cancer

Lung cancer is a leading cause of cancer-related death, about 40% human non-small cell lung cancer (NSCLC) patients showed lymph node involvements. However, the precise mechanism for the metastasis is still not fully understood. This study was to analyze the potential molecular mechanism for lung cancer metastasis. In the current study, proteomics analysis by two-dimensional electrophoresis (2-DE) was performed first to identify the differentially expressed protein between the higher metastasis lung adenocarcinoma cell line Anip973 and the lower metastasis lung adenocarcinoma cell line AGZY83-a. We confirmed the result by RT-PCR, immunoblotting and immunocytochemistry analyses in these two cell lines. Then we examined the expression of the differentially expressed protein in tumor tissues of NSCLC patients by immunoblotting and immunohistochemistry analyses. Using 2-DE analysis, we have identified DJ-1 was expressed higher in the higher metastasis Anip973 compared to the parental cell line AGZY83-a, that was confirmed by RT-PCR, immunoblotting and immunocytochemistry analyses. In NSCLC patients?? tumor tissues study, immunoblotting data showed that, DJ-1 expression level was significantly higher in 72.2% (13/18) of NSCLC tissue samples compared to that in paired normal lung tissues (P?=?0.044). Immunohistochemistry analysis demonstrated increased DJ-1 expression in 85 NSCLC tumor tissue samples compared with 7 normal lung tissue samples (P?=?0.044). DJ-1 expression was also found to be significantly correlated with cancer lymphatic metastasis (P?=?0.039). DJ-1 might contribute to the metastasis of NSCLC.     

Redd1 inhibits the invasiveness of non-small cell lung cancer cells

Redd1 acts as a negative regulator of mTOR in response to various stress conditions, but its specific physiological role is currently unclear. In the present study, we showed that Redd1 inhibits the invasive activity of non-small cell lung cancer (NSCLC) cells. Interestingly, expression of Redd1 was extremely low in H1299 cells displaying high invasiveness, compared with that in H460 cells with lower invasive activity. Overexpression of Redd1 inhibited the invasive activity of H1299 cells, while suppression with specific siRNAs enhanced the invasiveness of H460 cells. Knockdown of the mTOR downstream substrate, S6K, resulted in a decrease in the invasive property of H1299 cells. Our results provide preliminary evidence that Redd1 inhibits the invasive activity of NSCLC cells via suppression of the mTOR downstream pathway.     

Expression of the cell adhesion molecule CD146/MCAM in non-small cell lung cancer.

Expression of MCAM is observed in a variety of human malignancies. We aimed to determine the rate of MCAM expression in our non-small cell lung cancer (NSCLC) collection and to clarify its correlation with clinicopathological parameters and patient survival. 85 NSCLC were analysed immunohistochemically using a monoclonal MCAM antibody (clone N1238) on an NSCLC tissue micro array. The staining was semiquantitatively scored. We found MCAM expression in 51% of NSCLC, preferentially squamous cell carcinomas (p=0.004). No other correlations to tumour size, grade, or stage were found. Univariate survival analysis showed no significant differences of MCAM positive and negative tumours. In adenocarcinomas however, MCAM positivity was significantly associated with shorter patient survival (p=0.016). We conclude, that MCAM is expressed in a high proportion of NSCLC and might be predictive of shortened patient survival in adenocarcinomas of the lung. Colour figure can be viewed on http://www.esacp.org/acp/2003/25-2/kristiansen.htm     

Endothelin-1 is a novel prognostic factor in non-small cell lung cancer

Endothelin-1 (ET-1) is a potent vasoactive peptide and a hypoxia-inducible angiogenic growth factor associated with the development and growth of solid tumours. This study evaluated the expression of big endothelin-1 (big ET-1), a stable precursor of ET-1, and ET-1 in non-small cell lung cancer (NSCLC). Big ET-1 expression was evaluated in paraffin-embedded tissue sections from 10 NSCLC tumours using immunohistochemistry and in situ hybridisation. The production of big ET-1 and ET-1 was studied in six established NSCLC cell lines. The plasma concentrations of big ET-1 were measured in 30 patients with proven NSCLC prior to chemotherapy by means of a sandwich enzyme-linked immunoassay and compared to levels in 20 normal controls. Big ET-1 immunostaining was detected in the cancer cells of all tumours studied. Using in situ hybridisation, tumour cell big ET-1 mRNA expression was demonstrated in all samples. All six NSCLC cell lines expressed ET-1, with big ET-1 being detected in three. The median big ET-1 plasma level in patients with NSCLC was 5.4 pg/mL (range 0-22.7 pg/mL) and was significantly elevated compared to median big ET-1 plasma levels in controls, 2.1 pg/mL (1.2-13.4 pg/mL) (p = 0.0001). Furthermore, patients with plasma big ET-1 levels above the normal range (upper tertile) had a worse outcome (p = 0.01). In conclusion, big ET-1/ET-1 is expressed by resected NSCLC specimens and tumour cell lines. Plasma big ET-1 levels are elevated in NSCLC patients compared to controls with levels > 7.8 pg/mL being associated with a worse outcome. The development of selective ET-1 antagonists such as Atrasentan indicates that ET-1 may be a therapeutic target in NSCLC.     

p120ctn、Kaiso及matrilysin在非小细胞肺癌中的表达及其意义

目的探讨p120catenin(p120ctn)、Kaiso和matrilysin在非小细胞肺癌(non-small cell lung cancer,NSCLC)发生发展中的作用。方法采用免疫组织化学法检测98例NSCLC(52例肺鳞状细胞癌和46例肺腺癌)以及16例支气管扩张症组织中p120ctn、Kaiso和matrilysin的表达情况,分析NSCLC中p120ctn、Kaiso和matrilysin与临床病理特征的关系及三者表达的相关性。结果在NSCLC组织和支气管扩张症组织中,p120ctn蛋白异常表达率分别为74.5%、6.3%,两组之间表达率比较有统计学差异(P<0.001);在肺癌细胞、杯状细胞、纤毛细胞、基底细胞中Kaiso蛋白异位表达率分别为63.3%、6.3%、6.3%、37.5%,肺癌细胞与杯状细胞、纤毛细胞表达率比较有统计学差异(P<0.001)。p120ctn蛋白异常表达、matrilysin蛋白细胞质表达均与NSCLC组织学类型密切相关(P<0.05),Kaiso蛋白异位表达与NSCLC分化程度密切相关(P<0.05)。在NSCLC组织中,Kaiso细胞核表达与matrilysin细胞质表达存在显著负相关(r=-0.23,P=0.02)。结论 p120ctn可能通过与Kaiso在胞质内异位表达,解除了Kaiso对靶基因的抑制作用,促进matrilysin的细胞质表达,而参与肺癌的发生发展。