Immune Microenvironment Comparation Study between EGFR Mutant and EGFR Wild Type Lung Adenocarcinoma Patients Based on TCGA Database / 中国肺癌杂志

BACKGROUND@#Lung cancer is a malignant with high incidence and mortality and adenocarcinoma is among the most popular subtypes. Epidermal growth factor receptor (EGFR) mutation is one of the most important driver mutations for lung adenocarcinoma and EGFR-tyrosine kinase inhibitor (TKI) will benefit those patients with sensitive EGFR mutations. Recently, immune checkpoint inhibitor (ICI) therapy, provide a new breakthrough treatment for lung cancer patients. Whereas immunotherapy as an emerging treatment does not benefit patients with EGFR mutations, for which mechanistic studies are poorly defined and focused on the link of EGFR mutations and programmed cell death-ligand 1 (PD-L1) expression, we speculate that the different immune microenvironment associated with the two classes of patients.@*METHODS@#Lung adenocarcinoma datasets were collected from the Cancer Genome Atlas (TCGA) database, and clinical information and gene expression profiles were downloaded. The immune related lymphocyte infiltration in TCGA database were generated through timer 2.0 GSEA was used to analyze the difference of pathway expression between EGFR mutant patients and wild type patients.@*RESULTS@#EGFR mutation was more frequently among women and never smokers. Immunoinfiltration analysis showed that patients with EGFR mutation tends to have more tumor associated fibroblasts, common myeloid progenitor cells, hematopoietic stem cells, effector CD4⁺ T cells and natural killer T cells infiltration, and less memory B cells, naïve B cells, plasma B cells, plasmacytoid dendritic cells, memory CD4⁺ T cells, CD4⁺ helper T cells 2, naive CD8⁺ T cells, CD8⁺ T cells and central memory CD8⁺ T cells infiltration. Moreover, patients with more infiltration of CD8⁺ T cells, natural killer T cells, memory B cells and hematopoietic stem cells, tends have better prognosis (Log-rank test, P=0.017, 0.0093, 0.018, 0.016). However, the patients with more CD4⁺ T th2 infiltration in the tumor tends to have worse prognosis (Log-rank test, P=0.016). Furthermore, the results of gene set enrichment analysis showed that compared with the lung adenocarcinoma patients with EGFR wild type, the three pathways positive regulation of natural killer (NK) cell-mediated immune response to tumor cells, NK cell activation involved in immune response, and NK cell-mediated immune response to tumor cells related to natural killer cells in patients with EGFR mutation were down regulated, while the pathway the positive regulation of cytokine secretion involved in immune response was up-regulated in EGFR mutation patients.@*CONCLUSIONS@#The tumour microenvironment of patients with EGFR mutations lacks potent tumour killing effector cells and appears dysfunctional with effector cells. This may be a potential reason for the poor efficacy of immunotherapy in patients with EGFR mutations.

Apatinib Combined with CCI-779 Inhibits the Proliferation and Migration of Small Cell Lung Cancer NCI-H446 Cells In Vitro / 中国肺癌杂志

BACKGROUND@#Lung cancer is the most common malignancy world-wide. Small cell lung cancer is the deadliest subtype of lung cancer, which features such as rapid growth, early metastasis, and high vascularization. Apatinib is a vascular endothelial growth factor receptor 2 inhibitor independently developed in China, which has a significant inhibition in a variety of solid tumors. The purpose of this study is to investigate the effects of Apatinib alone or Apatinib combined with mammalian target of rapamycin (mTOR) inhibitor, CCI-779, on small cell lung cancer cell line NCI-H446 in vitro.@*METHODS@#The small cell lung cancer cell line NCI-H446 was grew in vitro. The effects of Apatinib alone or Apatinib combined with CCI-779 on proliferation, apoptosis, cell cycle and migration of NCI-H446 small cell lung cancer cells were detected by CCK8; FACS and transwell assays were also carried out; Western blot assays were used to detect vascular endothelial growth factor and cell cycle related protein expression.@*RESULTS@#CCK8 assays showed that high concentration of Apatinib could inhibit the proliferation of NCI-H446 cells. Apoptosis assays showed that high concentration of Apatinib could induce NCI-H446 cell apoptosis. Transwell assays showed that high concentration of Apatinib could inhibit NCI-H446 cell migration. After combined with mTOR inhibitor CCI-779, low concentration of Apatinib could inhibit the proliferation and migration of NCI-H446 small cell lung cancer cells and induce apoptosis.@*CONCLUSIONS@#Apatinib has a concentration-dependent effect on the small cell lung cancer cell line NCI-H446. High concentration of Apatinib can inhibit the proliferation and migration of NCI-H446 small cell lung cancer cells, induce apoptosis. Apatinib combined with the mTOR inhibitor CCI-779 can sensitize the NCI-H446 cells to Apatinib.

The Role of Plasma CDO1 Methylation in the Early Diagnosis of Lung Cancer / 中国肺癌杂志

BACKGROUND@#The incidence and mortality of lung cancer often rank first in all malignant tumors. DNA methylation, as one of epigenetics, often participates in the development and progression of tumors. CDO1 as a tumor suppressor gene always undergoes methylation changes early in tumor development. Therefore, this study aims to discuss the value of CDO1 methylation in the early diagnosis of lung cancer.@*METHODS@#Peripheral blood samples were collected from tumor patients and healthy people. Detection of the methylation level of CDO1 in plasma by sulfite modification and quantitative real-time PCR.@*RESULTS@#The level of gene methylation in peripheral blood of lung cancer patients was significantly higher than that of benign lung disease patients and healthy people. The methylation level of CDO1 was significantly different in the stratified comparison of gender, lymph node metastasis and tumor-node-metastasis (TNM) stage (P<0.05). The sensitivity and specificity of CDO1 were 52.2% and 78.6%, respectively. The overall accuracy of the diagnosis was significantly higher than that of the clinical tumor markers, and the sensitivity of CDO1 to stage I and II patients was the highest (40.8%, 47.1%). In addition, CDO1 could effectively increase the sensitivity of diagnosis in multiple joint examinations.@*CONCLUSIONS@#Detecting the methylation level of CDO1 has a potentially huge advantage for the early diagnosis of lung cancer.

Research Progress of Epigenetics in Pathogenesis and Treatment of Malignant Tumors / 中国肺癌杂志

Epigenetic modification is closely related to the occurrence and development of tumors. It mainly regulates gene function and expression level through DNA methylation, histone modification, regulation of non-coding RNA and chromatin structure reconstruction. At present, epigenetic drugs have been gradually applied to the treatment of malignant tumors. Common drug types include: DNA methyltransferase inhibitors and histone deacetylase inhibitors. However, these drugs still have many shortcomings and a wide range of clinical applications need further research. Encouragingly, the epigenetic drugs in combination with various anti-tumor drugs have shown great application potential. In this paper, we summarized the development mechanism of epigenetics in malignant tumors and the progress of related drugs.

Role of EZH2 Inhibitor Combined with Gefitinib in EGFR-TKIs Resistant Lung Cancer Cells / 中国肺癌杂志

BACKGROUND@#Lung cancer is one of the common malignant tumors that impair human health. With the development of epigenetics, the researchers found that enhancer of Zeste homolog 2 (EZH2) is highly expressed in lung cancer tissue and its expression is closely related to the prognosis. EZH2 inhibitor can also enhance the sensitivity of tumor cells to a variety of anti-tumor drugs. The purpose of this study is to investigate the effect of combination of EZH2 inhibitor and gefitinib on the proliferation, apoptosis and migration of Gefitinib-resistant lung cancer cells.@*METHODS@#PC9 and PC9/AB2 cells were used for this study. CCK-8 and EdU experiment were used to detect combined treatment on cell viability and proliferation activity; Wound healing assay and Transwell chamber experiment were used to determine the effects of combination therapy on cell migration ability; Flow cytometry was used to detect the effect of combination therapy on EZH2 and apoptosis; Western blot was used to observe the effect of combination therapy on epidermal growth factor receptor (EGFR) signaling pathway-related proteins expression.@*RESULTS@#In gefitinib-resistant cell line PC9/AB2, gefitinib combined with EZH2 inhibitor GSK343 can significantly inhibit cell viability, reduce cell migration and increase cell apoptosis. At the same time, combination therapy can significantly inhibit the expression of EZH2 and phosphorylation EGFR proteins.@*CONCLUSIONS@#The combination of EZH2 inhibitor GSK343 and gefitinib sensitize PC9/AB2 cell to gefitinib response. This study also suggests that synergistic therapy plays a role in the reversal of EGFR-tyrosine kinase inhibitor (EGFR-TKIs) resistance in lung cancer.

Effect of Apatinib on Invasion and Migration of Lung Cancer Cells and Its Mechanism / 中国肺癌杂志

BACKGROUND@#Lung cancer is one of the most deadly cancers in the world for human. In recent years, the effect of targeted therapy has become increasingly significant. Apatinib is a multi-target anti-tumor drug that is currently under study. The purpose of this study is to investigate the effects of Apatinib on the biological characteristics of lung cancer cells and its possible mechanism.@*METHODS@#Lung cancer cell lines H1299 and H3255 were cultured in vitro. The effects of Apatinib on proliferation, migration and invasion of H1299 and H3255 cells were detected by cell proliferation assays wound healing assays and Transwell assays. The protein expression related to cancer angiogenesis and invasion was detected by Western blot.@*RESULTS@#Apatinib significantly inhibited the proliferation, migration and invasion of H1299 and H3255 in a concentration-dependent manner. Western blot showed that with the increasing of drug concentration, VEGF, VEGFR2, N-cadherin, MMP9, MMP2 and Vimentin were down-regulated, and E-cadherin were up-regulated.@*CONCLUSIONS@#Apatinib can inhibit the invasion and migration of lung adenocarcinoma cells H1299 and H3255. By regulation of epithelial-mesenchymal transition and the expression of matrix metalloproteinase-related proteins.

RNF126 Quenches RNF168 Function in the DNA Damage Response / 基因组蛋白质组与生物信息学报·英文版

DNA damage response (DDR) is essential for maintaining genome stability and protecting cells from tumorigenesis. Ubiquitin and ubiquitin-like modifications play an important role in DDR, from signaling DNA damage to mediating DNA repair. In this report, we found that the E3 ligase ring finger protein 126 (RNF126) was recruited to UV laser micro-irradiation-induced stripes in a RNF8-dependent manner. RNF126 directly interacted with and ubiquitinated another E3 ligase, RNF168. Overexpression of wild type RNF126, but not catalytically-inactive mutant RNF126 (CC229/232AA), diminished ubiquitination of H2A histone family member X (H2AX), and subsequent bleomycin-induced focus formation of total ubiquitin FK2, TP53-binding protein 1 (53BP1), and receptor-associated protein 80 (RAP80). Interestingly, both RNF126 overexpression and RNF126 downregulation compromised homologous recombination (HR)-mediated repair of DNA double-strand breaks (DSBs). Taken together, our findings demonstrate that RNF126 negatively regulates RNF168 function in DDR and its appropriate cellular expression levels are essential for HR-mediated DSB repair.

MR-fluorescent dual-modality molecular probes anchored with LyP-1 for pancreatic cancer / 中国医学影像技术

Objective To construct the LyP-1 targeted MR fluorescence dual-modality molecular probe for pancreatic cancer,and to observe its features and MRI charicteristics.Methods The 50 nm MR-fluorescent dual-modality molecular probe with surface modified with cyclic nine-amino acid peptide LyP-1 (Cys-Gly-Asn-Lys-Arg-Thr-Arg-Gly Cys) was rationally designed.Whether the molecular probe could specifically recognize the pancreatic cancer cells were validated by the combination of fluorescent imaging and MR T2WI.Results The new MR-fluorescent dual-modality molecular probe anchored with LyP-1 could be used for the fluorescent imaging and MR T2WI of pancreatic cancer in mouse.And the molecular probe was demonstrated to be effective in conjugating with pancreatic cancer cells on fluorescent images and caused obvious MR signal reduction under T2 relaxometry in vitro.In vivo experiment,the molecular probe could be used for fluorescent labeling tumor tissue and detecting orthotopic pancreatic cancer in C57BL/6 mouse as MR contrast agent.Conclusion The LyP1 immobilized MR-fluorescent dual-modality molecular probe can actively target to mouse orthotopic xenograft of pancreatic cancer,which is hopeful to the application in early probing and diagnosis of pancreatic cancer by multimodal imaging.

MR-fluorescent dual-modality molecular probes anchored with LyP-1 for pancreatic cancer / 中国医学影像技术

Objective To construct the LyP-1 targeted MR fluorescence dual-modality molecular probe for pancreatic cancer,and to observe its features and MRI charicteristics.Methods The 50 nm MR-fluorescent dual-modality molecular probe with surface modified with cyclic nine-amino acid peptide LyP-1 (Cys-Gly-Asn-Lys-Arg-Thr-Arg-Gly Cys) was rationally designed.Whether the molecular probe could specifically recognize the pancreatic cancer cells were validated by the combination of fluorescent imaging and MR T2WI.Results The new MR-fluorescent dual-modality molecular probe anchored with LyP-1 could be used for the fluorescent imaging and MR T2WI of pancreatic cancer in mouse.And the molecular probe was demonstrated to be effective in conjugating with pancreatic cancer cells on fluorescent images and caused obvious MR signal reduction under T2 relaxometry in vitro.In vivo experiment,the molecular probe could be used for fluorescent labeling tumor tissue and detecting orthotopic pancreatic cancer in C57BL/6 mouse as MR contrast agent.Conclusion The LyP1 immobilized MR-fluorescent dual-modality molecular probe can actively target to mouse orthotopic xenograft of pancreatic cancer,which is hopeful to the application in early probing and diagnosis of pancreatic cancer by multimodal imaging.

Histopathology of the temporal artery in young ischemic stroke patients with elevated serum immunoglobu-lin E / 医学研究生学报

Objective Ischemic stroke with elevated serum immunoglobulin E ( IgE) in some young patients is regarded as cerebral vasculitis clinically though without sufficient pathological evidence .This study was to investigate the characteristics of vascular lesions in these patients by temporal artery biopsy . Methods We performed histopathologic examinations on the temporal arteries of 32 young ischemic stroke patients with unknown etiology , 16 with normal and the other 16 with elevated serum IgE .We observed inflammatory cells infiltration and mast cells by HE staining and toluidine blue stai-ning respectively and determined the expressions of matrix metalloproteinase -9 (MMP-9), monocyte chemotaxis protein -1 (MCP-1) and serum IgE by immunohistochemistry . Results Compared with the patients with normal IgE , those of the elevated IgE group showed a significantly higher rate of inflammatory cells infiltration (12.5%vs 62.5%, P0.05).Nor was any signifi-cant difference observed in the number of the mast cells between the normal and elevated IgE groups (2.8 ±1.5 vs 3.6 ±2.3, P>0.05). Conclusion The infiltration and necrosis of inflammatory cells and fibrin exudation in the temporal artery of the young pa-tient with elevated serum IgE are likely to be the manifestations of vasculitis , and MCP-1 may play a role in the pathogenesis of the disease.

Predictive value of leukocyte differential count in patients with acute cerebral infarction / 医学研究生学报

Objective Inflammation response is involved in the whole pathological process of acute cerebral infarction ( ACI) , but few reports are seen on its clinical implication in ACI patients .The purpose of this study was to investigate the predictive value of the differential count of leukocytes for stroke severity and early clinical outcomes in the acute phase of cerebral infarction . Methods We collected the clinical and laboratory data of 635 patients diagnosed with ACI within 72 hours of symptom onset and eval-uated the association between the differential count of peripheral blood leukocytes and stroke severity at admission and within 3 days af-ter admission as well as the clinical outcomes at discharge .The neural function impairment scores of the patients were obtained with The NIH Stroke Score ( NIHSS) at admission and on the third day after admission , and the therapeutic results evaluated with the modi-fied Rankin Scale ( mRS) , mRS >2 as poor prognosis .Analyses were performed on the correlation of the differential count of leuko-cytes with NIHSS and mRS scores and its influence on the ACI patients . Results At discharge , the mRS related influencing factors included the total count of leukocytes (OR=1.147, 95% CI:1.038-1.268), count of neutrophil cells (OR=1.227, 95% CI:1.00-1.369 ), count of lymphocytes ( OR =0.508, 95% CI:0.342-0.753), and neutrophil to lymphocyte ratio (NLR) (OR=1.150, 95%CI:1.008-1.314).the NIHSSs were correlated with the counts of leucocytes (r=0.078, P=0.024), neutrophil cells (r=0.083, P=0.019), and lymphocytes (r=0.010, P=0.004) at admission, and with the counts of leucocytes ( r =0.238, P <0.001), neutrophil cells (r=0.335, P<0.001), lymphocytes (r=-0.269, P<0.001), and NLR (r=0.423, P<0.001) on the third day after admission. Conclusion In the acute phase of cer-ebral infarction , the differential count of leukocytes and NLR are valuable for predicting the severity of neurologic impairment and early poor functional outcome .

Correlation of previous treatment of hypertension with the severity of ischemic stroke / 医学研究生学报

Objective Hypertension is a leading modifiable risk factor for cardiovascular disease .However , a lot of hyper-tension patients hold inactive attitudes to hypertension treatment .The purpose of this study was to investigate the relationship between previous treatment of hypertension and stroke severity in acute ischemic stroke . Methods We retrospectively analyzed the clinical data of 653 in-hospital ischemic stroke patients with hypertension between January 2011 and December 2014 .According to the National Institutes of Health Stroke Scale (NIHSS) at admission, the stroke patients were divided into a mild group (NIHSS≤3) and a severe group (NIHSS >3) and, based on their history of hypertension treatment , allocated to a regular treatment, an irregular treatment, a non-treatment , and an unawareness group .We studied the relationship of previous hypertension treatment with stroke severity by Spearman correlation analysis and identified the potential factors associated with stroke severity by multivariate logistic regression anal-ysis. Results Previous treatment of hypertension was positively correlated with stroke severity (r=0.146, P=0.000 2).Compared with the patients of the regular treatment group , those in the irregular treatment group (OR: 2.21; 95%CI:1.39 -3.52; P =0.001), non-treatment group ( OR: 2.18; 95%CI: 1.41 -3.36; P =0.0004) and unawareness group (OR:1.80;95%CI:1.12-2.88; P=0.015) tended to have more severe stroke. Conclusion Previous treatment of hypertension is closely related to the severity of ischemic stroke .

Protective effect of tissue kallikrein against inflammation following focal cerebral ischemia-reperfusion in diabetic rats / 医学研究生学报

Objective It remains to be confirmed whether tissue kallikrein has neuroprotective effect in diabetes-induced stroke.This study was to investigate the neuroprotection of tissue kallikrein against cerebral ischemia-reperfusion injury in diabetic rats. Methods Healthy male SD rats were randomly divided into a sham operation, a saline control, and a tissue kallikrein group.Diabetes mellitus was induced in the animals by intraperitoneal injection of streptozotocin and the model of focal cerebral ischemia-reperfusion was made with an intraluminal vascular occlusion method. At 24 hours after modeling, we obtained the neurological deficit score, in-farct size, and brain water content, counted Iba1-and MPO-positive cells by immunohistochemistry, and determined the expressions of ICAM-1 and VCAM-1 by real-time PCR. Results In comparison with the saline controls, the rats treated with tissue kallirein showed significant decreases in the neurological deficit score (P<0.01), the infarct size ([23.57 ±5.79] vs [47.97 ±1.19]%, P<0.01), brain edema ([81.73 ±2.10] vs [84.94 ±2.34]%, P<0.05), the counts of Iba1-and MPO-positive cells (12.33 ±4.46 vs 31.83 ±8.13 and 13.83 ±4.49 vs 37.50 ±7.64, both P<0.01), and the expressions of ICAM-1 and VCAM-1 (both P<0.05). Conclusion Tissue kallikrein has a neuroprotective effect against cerebral ischemia-reperfusion injury in diabetic rats, which may be associated with its anti-inflammation property.

Research progress on the intracellular mechanisms of kallikrein-kinin system / 医学研究生学报

Kallikrein-kinin system consists of kininogen , kallikrein, bradykinin and kinin .Kinins, derived from kininogen by tissue kallikrein , play their biological effects via bradykinin 1/2 receptors or protease activated receptors .Existing researches suggest that kinins exert various effects through different intracellular and mitochondrial signal pathways such as MAPK , PI3K/Akt/GSK3 be-ta, NO, JAKs/STATs.This review aims to elucidate the roles and the intracellular signal pathways of KKS in different diseases .