Cell Differentiation Of Lung Cancer Research Articles

Application of Inorganic Nanomaterials for Drug Delivery in Lung Cancer

Lung cancer has been one of the most common cancers in the world for more than 100 years and has taken many people's lives. Many people suffer from lung cancer due to smoking, second-hand smoke, and serious air pollution or pollutant gathering environment. It is necessary to find a good medical solution. Compared with traditional chemotherapy and immunotherapy, targeted therapy has more advantages in the impact on patients and the efficiency of treatment. How to develop targeted therapeutic drugs has become a topic, and nanoscale targeted drugs have become the most appropriate choice. Among them, there are many inorganics suitable for nano-targeted drugs and nano-drug carriers, such as metals, metal oxides, and non-metals. By blocking the genetic material in lung cancer cells, it can avoid the division and differentiation of lung cancer cells, so as to achieve the purpose of treating lung cancer. This review will focus on the above three kinds of nano-targeted drugs, and discuss their therapeutic principles, drug delivery, activation mode, and biotoxicity.

Neuroendocrine Differentiation of Lung Cancer Cells Impairs the Activation of Antitumor Cytotoxic Responses in Mice.

Lung cancer has the highest mortality among all types of cancer; during its development, cells can acquire neural and endocrine properties that affect tumor progression by releasing several factors, some acting as immunomodulators. Neuroendocrine phenotype correlates with invasiveness, metastasis, and low survival rates. This work evaluated the effect of neuroendocrine differentiation of adenocarcinoma on the mouse immune system. A549 cells were treated with FSK (forskolin) and IBMX (3-Isobutyl-1-methylxanthine) for 96 h to induce neuroendocrine differentiation (NED). Systemic effects were assessed by determining changes in circulating cytokines and immune cells of BALB/c mice immunized with PBS, undifferentiated A549 cells, or neuroendocrine A549NED cells. A549 cells increased circulating monocytes, while CD4+CD8- and CD4+CD8+ T cells increased in mice immunized with neuroendocrine cells. IL-2 and IL-10 increased in mice that received untreated A549 cells, suggesting that the immune system mounts a regulated response against adenocarcinoma, which did not occur with A549NED cells. Cocultures demonstrated the cytotoxic capacity of PBMCs when confronted with A549 cells, while in the presence of neuroendocrine cells they not only were unable to show cytolytic activity, but also lost viability. Neuroendocrine differentiation seems to mount less of an immune response when injected in mice, which may contribute to the poor prognosis of cancer patients affected by this pathology.

Elevation of EIF4G1 promotes non‐small cell lung cancer progression by activating mTOR signalling

Eukaryotic translation initiation factor 4 gamma 1 (EIF4G1), as the key component of the transcription initiation factor complex EIF4F, is significantly upregulated in multiple solid tumours, including lung cancer. However, the function and mechanism of EIF4G1 in the regulation of non‐small‐cell lung cancer (NSCLC) remain unclear. Here, using the clinical samples and the comprehensive survival analysis platforms Kaplan‐Meier plotter, we observed aberrant upregulation of EIF4G1 in NSCLC tissues; furthermore, high expression of EIF4G1 showed association with low differentiation of lung cancer cells and poor overall survival in NSCLC patients. Non‐small‐cell lung cancer cell line A549 and H1703 stably infected with EIF4G1 shRNA were used to determine the function of EIF4G1 in regulating cell proliferation and tumorigenesis in vitro and in vivo. The results demonstrated that EIF4G1 promoted the G1/S transition of the cell cycle and tumour cell proliferation in non‐small cell lung cancer. Mechanistically, EIF4G1 was found to regulate the expression and phosphorylation of mTOR (Ser2448), which mediates the tumorigenesis‐promoting function of EIF4G1. The inhibition of mTOR attenuated the EIF4G1‐induced development and progression of tumours. These findings demonstrated that EIF4G1 is a new potential molecular target for the clinical treatment of non‐small cell lung cancer.

Immunohistochemical quantification of expression of a tight junction protein, claudin-7, in human lung cancer samples using digital image analysis method

Background and objectivesTight junction proteins are correlated with cancer development. As the pivotal proteins in epithelial cells, altered expression and distribution of different claudins have been reported in a wide variety of human malignancies. We have previously reported that claudin-7 was strongly expressed in benign bronchial epithelial cells at the cell-cell junction while expression of claudin-7 was either altered with discontinued weak expression or completely absent in lung cancers. Based on these results, we continued working on the expression pattern of claudin-7 and its relationship with lung cancer development. We herein proposed a new Digital Image Classification, Fragmentation index, Morphological analysis (DICFM) method for differentiating the normal lung tissues and lung cancer tissues based on the claudin-7 immunohistochemical staining. MethodsSeventy-seven lung cancer samples were obtained from the Second Affiliated Hospital of Zhejiang University and claudin-7 immunohistochemical staining was performed. Based on C++ and Open Source Computer Vision Library (OpenCV, version 2.4.4), the DICFM processing module was developed. Intensity and fragmentation of claudin-7 expression, as well as the morphological parameters of nuclei were calculated. Evaluation of results was performed using Receiver Operator Characteristic (ROC) analysis. ResultsAgreement between these computational results and the results obtained by two pathologists was demonstrated. The intensity of claudin-7 expression was significantly decreased while the fragmentation was significantly increased in the lung cancer tissues compared to the normal lung tissues and the intensity was strongly positively associated with the differentiation of lung cancer cells. Moreover, the perimeters of the nuclei of lung cancer cells were significantly greater than that of the normal lung cells, while the parameters of area and circularity revealed no statistical significance. ConclusionsTaken together, our DICFM approach may be applied as an appropriate approach to quantify the immunohistochemical staining of claudin-7 on the cell membrane and claudin-7 may serve as a marker for identification of lung cancer.

Econazole nitrate inhibits PI3K activity and promotes apoptosis in lung cancer cells

The phosphatidylinositol-3-kinase (PI3K)/AKT signaling pathway plays a pivotal role in many cellular processes, including the proliferation, survival and differentiation of lung cancer cells. Thus, PI3K is a promising therapeutic target for lung cancer treatment. In this study, we applied free and open-source protein-ligand docking software, screened 3167 FDA-approved small molecules, and identified putative PI3Kα inhibitors. Among them, econazole nitrate, an antifungal agent, exhibited the highest activity in decreasing cell viability in pathological types of NSCLC cell lines, including H661 (large cell lung cancer) and A549 (adenocarcinoma). Econazole decreased the protein levels of p-AKT and Bcl-2, but had no effect on the phosphorylation level of ERK. It inhibited cell growth and promote apoptosis in a dose-dependent manner. Furthermore, the combination of econazole and cisplatin exhibited additive and synergistic effects in the H661 and A549 lung cancer cell lines, respectively. Finally, we demonstrated that econazole significantly suppressed A549 tumor growth in nude mice. Our findings suggest that econazole is a new PI3K inhibitor and a potential drug that can be used in lung cancer treatment alone or in combination with cisplatin.

Class III/IV POU transcription factors expressed in small cell lung cancer cells are involved in proneural/neuroendocrine differentiation.

One-third of lung malignancies demonstrate a proneural/neuroendocrine phenotype or type of differentiation. However, it has not been clearly elucidated how proneural/neuroendocrine differentiation is controlled in lung cancers. We recently demonstrated that the POU3F2 gene plays a significant role in proneural/neuroendocrine differentiation of lung cancers. Because class III POU genes (POU3F1, POU3F2, POU3F3, and POU3F4) and class IV POU genes (POU4F1, POU4F2, and POU4F3) share similar properties in neural development, we analyzed the association between class III/IV POU genes and a proneural/neuroendocrine phenotype in lung cancers using seven small cell lung cancer (SCLC) cell lines and twelve non-SCLC (NSCLC) cell lines. Class III/IV POU gene expression was generally restricted to SCLC cells. However, the forced expression of class III/IV POU genes in the NSCLC cell lines induced the expression of neuroendocrine-specific markers (neural call adhesion molecule 1, synaptophysin, and chromogranin A) and proneural transcription factors (achaete-scute homolog-like 1, NeuroD1, and thyroid transcription factor 1) in various degrees. Furthermore, each class III/IV POU gene induced other class III/IV POU genes, suggesting the mutual induction of class III/IV POU genes. These findings suggest that the expression of class III/IV POU genes is important for the proneural/neuroendocrine differentiation of lung cancer cells.

Transforming growth factor-beta signaling network regulates plasticity and lineage commitment of lung cancer cells

Identification of target cells in lung tumorigenesis and characterization of the signals that control their behavior is an important step toward improving early cancer diagnosis and predicting tumor behavior. We identified a population of cells in the adult lung that bear the EpCAM+CD104+CD49f+CD44+CD24loSCA1+ phenotype and can be clonally expanded in culture, consistent with the properties of early progenitor cells. We show that these cells, rather than being restricted to one tumor type, can give rise to several different types of cancer, including adenocarcinoma and squamous cell carcinoma. We further demonstrate that these cells can be converted from one cancer type to the other, and this plasticity is determined by their responsiveness to transforming growth factor (TGF)-beta signaling. Our data establish a mechanistic link between TGF-beta signaling and SOX2 expression, and identify the TGF-beta/SMAD/SOX2 signaling network as a key regulator of lineage commitment and differentiation of lung cancer cells.

Involvement of TRPC Channels in Lung Cancer Cell Differentiation and the Correlation Analysis in Human Non-Small Cell Lung Cancer

The canonical transient receptor potential (TRPC) channels are Ca2+-permeable cationic channels controlling the Ca2+ influx evoked by G protein-coupled receptor activation and/or by Ca2+ store depletion. Here we investigate the involvement of TRPCs in the cell differentiation of lung cancer. The expression of TRPCs and the correlation to cancer differentiation grade in non-small cell lung cancer (NSCLC) were analyzed by real-time PCR and immunostaining using tissue microarrays from 28 patient lung cancer samples. The association of TRPCs with cell differentiation was also investigated in the lung cancer cell line A549 by PCR and Western blotting. The channel activity was monitored by Ca2+ imaging and patch recording after treatment with all-trans-retinoic acid (ATRA). The expression of TRPC1, 3, 4 and 6 was correlated to the differentiation grade of NSCLC in patients, but there was no correlation to age, sex, smoking history and lung cancer cell type. ATRA upregulated TRPC3, TRPC4 and TRPC6 expression and enhanced Ca2+ influx in A549 cells, however, ATRA showed no direct effect on TRPC channels. Inhibition of TRPC channels by pore-blocking antibodies decreased the cell mitosis, which was counteracted by chronic treatment with ATRA. Blockade of TRPC channels inhibited A549 cell proliferation, while overexpression of TRPCs increased the proliferation. We conclude that TRPC expression correlates to lung cancer differentiation. TRPCs mediate the pharmacological effect of ATRA and play important roles in regulating lung cancer cell differentiation and proliferation, which gives a new understanding of lung cancer biology and potential anti-cancer therapy.

Modulation of endoplasmic reticulum calcium pump expression during lung cancer cell differentiation

Cellular calcium signaling plays important roles in several signal transduction pathways that control proliferation, differentiation and apoptosis. In epithelial cells calcium signaling is initiated mainly by calcium release from endoplasmic-reticulum-associated intracellular calcium pools. Because calcium is accumulated in the endoplasmic reticulum by sarco/endoplasmic reticulum calcium ATPases (SERCA), these enzymes play a critical role in the control of calcium-dependent cell activation, growth and survival. We investigated the modulation of SERCA expression and function in human lung adenocarcinoma cells. In addition to the ubiquitous SERCA2 enzyme, the SERCA3 isoform was also expressed at variable levels. SERCA3 expression was selectively enhanced during cell differentiation in lung cancer cells, and marked SERCA3 expression was found in fully differentiated normal bronchial epithelium. As studied by using a recombinant fluorescent calcium probe, induction of the expression of SERCA3, a lower calcium affinity pump, was associated with decreased intracellular calcium storage, whereas the amplitude of capacitative calcium influx remained unchanged. Our observations indicate that the calcium homeostasis of the endoplasmic reticulum in lung adenocarcinoma cells presents a functional defect due to decreased SERCA3 expression that is corrected during pharmacologically induced differentiation. The data presented in this work show, for the first time, that endoplasmic reticulum calcium storage is anomalous in lung cancer cells, and suggest that SERCA3 may serve as a useful new phenotypic marker for the study of lung epithelial differentiation.

The significance of MAGED4 expression in non-small cell lung cancer as analyzed by real-time fluorescence quantitative PCR

The aim of this study was to detect differences in the expression levels of melanoma-associated antigen D4 (MAGED4) mRNA between non-small cell lung cancer (NSCLC) tissues and normal tissues, and to compare differences in the expression levels of MAGED4 in tumor patients. Patients were grouped according to age, gender, smoking history, tumor size, pathological classification, degree of lung cancer cell differentiation and presence of lymph node metastasis. The expression levels of MAGED4 were detected using real-time fluorescence quantitative PCR. MAGED4 expression was higher in squamous cell carcinomas compared to adenocarcinomas (P<0.05), in poorly differentiated tissues compared to well-differentiated tissues (P<0.05), and in patients with lymph node metastasis compared to patients without lymph node metastasis (P<0.05). MAGED4 may be used as a specific antigen for NSCLC to influence the improvement of diagnosis, prognosis and immunological therapy outcomes in lung cancer patients.

Expression profiles of thioredoxin family proteins in human lung cancer tissue: correlation with proliferation and differentiation

Lung cancer is one of the most common causes of cancer lethality worldwide. Despite recent progress, long-term survival remains poor. The aim of this study was to explore the expression pattern of the thioredoxin superfamily of proteins as potential new diagnostic and/or predictive markers. The expression of thioredoxin 1 (Trx1), thioredoxin reductase 1 (TrxR1), the isoforms TrxR1-v.2,3,5, glutaredoxin 1 (Grx1) and glutaredoxin 2 (Grx2) was examined by immunohistochemistry on paraffin-embedded sections from 42 cases of non-small cell lung cancer patients. Additional cases of lung cancer from tissue microarray were examined and the immunoreactivity was compared. All proteins except TrxR1 showed a significant correlation with the degree of differentiation in adenocarcinoma. Trx1 and TrxR1-v.2,3,5 also showed a significant correlation with differentiation in squamous carcinoma. Furthermore, Grx1 and Grx2 showed a clear inverse correlation with proliferation. The proliferation rate was further analysed in vitro in stably transfected Grx2 overproducing cells, showing that the proliferative effect of Grx2 is strictly dependent on subcellular localization. The thioredoxin family of proteins is important for growth and differentiation of lung cancer cells. The correlation with differentiation and proliferation of these enzymes makes them promising predictive/diagnostic markers.

Expressions of Ezrin protein (Ezrin) in lung cancer and its clinical implication

Objective To detect the expression of Ezrin (a member of ERM family) in lung cancer and its relation with invasion and metastasis of lung cancer cells. Methods Immunohistochemical staining was used to detect the expression of Ezfin in 45 cases of lung cancer tissue,23 cases of benign tumor tissue,and 12 cases of normal lung tissue. The relationship between the expression of Ezrin and clin-icopathologic features of lung cancer was analyzed. Results None of the normal intestinal tissues and be-nign tumor tissue were positive for Ezrin. In 45 lung cancer specimens, the positive expression rate of Ezrin was 61.3% (27/45). The Ezrin expression was correlated with the differentiation and metastasis of lung cancer cells (P<0.05),but not with age,sex and tumor size. Conclusion The expression of Ezrin in tumor tissue may be a new marker of the biological behaviors of lung cancer. Key words: Lung carcinoma; Ezrin; Immunohistochemical; Metastasis

Role of diffusion-weighted magnetic resonance imaging for predicting of tumor invasiveness for clinical stage IA non-small cell lung cancer☆

Recently, diffusion-weighted MR imaging (DWI) for the whole body has become available for clinical use, as has been previously used for the central nervous system. Favorable results have been reported using this imaging system to differentiate between benign and malignant lesions in some organs, and to correlate with the degree of cell differentiation in lung cancer. The purpose of this study was to assess the role of DWI for predicting tumor invasiveness of non-small cell lung cancers (NSCLC), especially for clinical stage IA patients. From January 2006 to September 2007, preoperative DWI and 18F-FDG-PET/CT were performed on 41 patients with clinical stage IA NSCLC who had undergone curative operations. Lung cancers that exhibited nodal, lymphovascular or pleural invasion were defined as invasive lung cancers. Nodules with strong dark signal, as observed by DWI in spinal cords, were defined as DWI-positive. We analyzed the associations between the pathological findings and the following preoperative clinical factors: age, gender, smoking history, preoperative CEA levels (<5.0 or >/=5.0ng/ml), preoperative tumor size, SUV max on PET/CT (<5.0 or >/=5.0) and DWI (positive or negative). A total of 15 lesions (37%) were assessed as DWI-positive and 26 lesions (63%) were DWI-negative. Univariate analyses showed positive correlations for development of invasive cancer with the preoperative CEA level (p=0.049), SUV max (p=0.001) and DWI (p<0.001). Multivariate analysis showed that DWI (p=0.005) was an independent predictive factor for tumor invasiveness. Our results suggest that DWI might be a useful method for predicting tumor invasiveness for clinical stage IA NSCLC.

Prognostic Significance of Neuroendocrine Differentiation in Non-small Cell Lung Cancer Patients' Survival

Objective: To observe the relationship between non-small cell lung cancer with neuroendrocrine differentiation (NSCLC-NE) and patients' postoperative survival. Methods: During April 1997 to April 1999, 98 cases of lung cancer were surgically treated. The tumor specimens of the patients were stained by NE markers, i.e. neuron specific enolase (NSE) and synaptophysin (SY). The intensity of NE markers reaction was divided as "+", "++", "+++" scale groups. The same specimens were also examined under an electron microscope for the specific neuroendocrine granules. All enrolled patients were followed up for 36 months, and the longest follow-up time was 60 months. The COX proportional hazard model multivariate analysis was applied to observe the relationship between the NSCLE-NE and the patients' postoperative survival. Results: In 91 cases of NSCLC, 63.7% (58/91) were positive for NE stain reaction. Among them, 59.3% (54/91) were positive for NSE and 24.1% (22/91) for SY. 48.4% (44/91) were considered as NSCLC-NE by the combination of NE marker stain reaction and electron microscopic examination. COX proportional hazard model multivariate analysis showed that the NSCLC-NE patients' survival was significantly shortened (P=0.048). The following factors were related to NSCLC-NE patients' survival: lung cancer cell differentiation (P=0.006), clinical lung cancer stage (P=0.001), the NE markers reaction (P=0.054). Conclusion: NSCLE-NE is significantly related to the cancer cell differentiation and the patients' postoperative survival. The NE markers should be applied clinically as one of prognostic factors to evaluate the postoperative survival of NSCLC patients.

Prognostic significance of neuroendocrine differentiation in non-small cell lung cancer patients' survival

To observe the relationship between non-small cell lung cancer with neuroendocrine differentiation (NSCLC-NE) and patients' postoperative survival. During April 1997 to April 1999, ninety-eight patients with lung cancer who were surgical treated were enrolled in this study. The tumor specimens were stained by NE markers: neuron specific enolase (NSE) and synaptophysin (SY). The NE marker reactions were divided as +, ++ and +++ scale groups. The same specimens were also examined by electron microscope for the specific neuroendocrine granule. All enrolled patients were followed up to 36 months, and the maximum follow-up time was 60 months. The Cox proportional hazard model multivariate analysis was applied to observe the relationship between the NSCLC-NE and the patients' postoperative survival. Ninety-one cases were diagnosed as NSCLC. In these patients, 63.7% (58/91) were positive for NE stain. Among them, 59.3% (54/91) were NSE-positive, and 24.1%(22/91) were SY-positive. The specific NE granules were found by the electron microscope examination in 33.0%(30/91). Combination of NE marker stain reaction and electronic microscope examination results, 48.4% (44/91) were considered as NSCLC-NE. The Cox proportional hazard model multivariate analysis showed that the NSCLC-NE patients' survival was significantly reduced (P=0.048). The following factors were related to NSCLC-NE patients' survival: lung cancer cell differentiation (P=0.006), stages of the cancer (P=0.001), and the NE markers reaction (P=0.054). NSCLC-NE is significantly related to the cancer cell differentiation and the patients' postoperative survival. The NE markers should be applied clinically as one of prognostic factors to evaluate the postoperative survival for NSCLC patients.

Expressions of matrix metalloproteinase 9 and tissue inhibitor of metalloproteinase 1 and their correlation with metastasis and prognosis in lung cancer

To investigate the expressions of matrix metalloproteinase 9 (MMP-9) and tissue inhibitor of metalloproteinase 1 (TIMP-1) and their correlation with metastasis and prognosis in human lung cancer. Immunohistochemical S-P method was used to detect the expression of MMP-9 and TIMP-1 in 65 lung cancer tissues, 35 hyperplastic and dysplastic epithelium from patients with non-cancerous pulmonary diseases, and 30 normal epithelial tissues of the lung. The positive expression rates of MMP-9 in normal tissue, hyperplastic or dysplastic epithelium, and lung cancer tissue were 16.7%(5/30), 42.9%(15/35) and 72.3%(47/65) respectively, whereas the positive rates of TIMP-1 expression in normal tissue, hyperplastic or dysplastic epithelium, and lung cancer tissue were 6.7%(2/30), 28.6%(10/35) and 50.8%(33/65) respectively. Significant differences of the expression rates of MMP-9 and TIMP-1 were found between lung cancer and normal groups, between lung cancer and hyperplasia groups, and between hyperplasia and normal groups (P < 0.05). Small cell carcinoma and adenocarcinoma had higher MMP-9 expression than squamous cell carcinoma (P < 0.025). Expression rate of MMP-9 had negative relation with cell differentiation of lung cancer (P < 0.05), and positive relation with TNM stage (P < 0.025). Between the survival time < 2 years group and ≥2 years groups, both the expressions of MMP-9 and TIMP-1 had significant difference (P < 0.05 ). The expression of MMP-9 was closely related to metastasis of lung cancer (P < 0.005), but the expression of TIMP-1 was not related to metastasis. Overexpression of MMP-9 may appear in precancerous lesion and at the early stage of lung cancer. Activation of MMP-9 gene may be an important factor for oncogenesis of the lung. MMP-9 and TIMP-1 may play important roles in lung cancer invasion and metastasis, their overexpression could act as a reference to evaluate metastasis and unfavourable prognosis of lung cancer.

Expression of endostatin and its relationship with the clinical pathophysiological characteristics in non-small cell lung cancer

To study the relationship between expression of endostatin and clinical and pathophysiological characteristics in the non-small cell lung cancer (NSCLC). The expression of endostatin was detected in 46 lung cancer tissues and paracancerous lung tissues, 14 benign pulmonary lesion tissues as control by immunohistochemical staining (LSAB method). The expression of endostatin in lung cancer tissues (84.91%±7.65%) was significantly higher than that in paracancerous tissues (63.70%±12.45%) and benign pulmonary lesion tissues (40.29%±15.01%) (P < 0.01); The expression of endostatin was closely related to the size of primary tumor, distant metastasis of the cancer, P-TNM stages and cell differentiation of lung cancer (P < 0.05), but not to the histological classification, lymph node status, location of the tumor, smoking or not, age and sex of the patients with lung cancer (P > 0.05). The expression of endostatin in NSCLC cancer tissues might be helpful to evaluate the biological behavior of lung cancer.