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1.
Acta Cytol ; 65(5): 393-402, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34343999

RESUMO

OBJECTIVE: The aim of the study was to investigate the mutation status of multiple driver genes by RT-qPCR and their significance in advanced lung adenocarcinoma using cytological specimens. MATERIALS AND METHODS: 155 cytological specimens that had been diagnosed with lung adenocarcinoma in the Fourth Hospital of Hebei Medical University were selected from April to November 2019. The cytological specimens included serous cavity effusion and fine-needle aspiration biopsies. Among cytological specimens, 108 cases were processed by using the cell block method (CBM), and 47 cases were processed by the disposable membrane cell collector method (MCM) before DNA/RNA extraction. Ten drive genes of EGFR, ALK, ROS1, BRAF, KRAS, NRAS, HER2, RET, PIK3CA, and MET were combined detected at one step by the amplification refractory mutation system and ABI 7500 RT-qPCR. RESULTS: The purity of RNA (p = 0.005) and DNA (p = 0.001) extracted by using the MCM was both significantly higher than that extracted by using the CBM. Forty-seven cases of fresh cell specimens processed by the MCM all succeeded in multigene detections, while of 108 specimens processed by the CBM, 6 cases failed in multigene detections. Among 149 specimens, single-gene mutation rates of EGFR, ALK, ROS1, RET, HER2, MET, KRAS, NRAS, BRAF, and PIK3CA mutations were 57.71%, 6.04%, 3.36%, 2.68%, 2.01%, 2.01%, 1.34%, 0.67%, 0% and 0% respectively, and 6 cases including 2 coexistence mutations. We found that mutation status was correlated with gender (p = 0.047), but not correlated with age (p = 0.141) and smoking status (p = 0.083). We found that the EGFR mutation status was correlated with gender (p = 0.003), age (p = 0.015) and smoking habits (p = 0.007), and ALK mutation status was correlated with age (p = 0.002). CONCLUSION: Compared with the CBM, the MCM can improve the efficiency of DNA/RNA extraction and PCR amplification by removing impurities and enriching tumor cells. And we speculate that the successful detection rate of fresh cytological specimens was higher than that of paraffin-embedded specimens. EGFR, ALK, and ROS1 mutations were the main driver mutations in patients with advanced lung adenocarcinoma. We speculate that EGFR and ALK are more prone to concomitant mutations, respectively. Targeted therapies for patients with coexisting mutations need further study.


Assuntos
Adenocarcinoma/genética , Adenocarcinoma/patologia , Biomarcadores Tumorais/genética , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Adulto , Idoso , Quinase do Linfoma Anaplásico/genética , Biópsia por Agulha Fina , Humanos , Masculino , Pessoa de Meia-Idade , Proteínas Tirosina Quinases/genética , Proteínas Proto-Oncogênicas/genética
3.
Acta Cytol ; 65(3): 235-241, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33631757

RESUMO

PURPOSE: For anaplastic lymphoma kinase (ALK) gene detection, the centrifugal sedimentation method (CSM) and cell block method (CBM) are commonly used to process samples of bloody pleural effusions (BPEs). However, in practice, the impurity content in the processed samples often affects the results and even leads to the detection failure. The purpose of this study was to establish a cell enrichment method (CEM) by using a disposable membrane cell collector to remove blood and inflammatory cells and enrich lung adenocarcinoma cells in BPE for more efficient RNA extraction and ALK gene detection. MATERIALS AND METHODS: CEM proposed in this study and the traditional CSM and CBM were used to treat BPE samples collected from 37 lung adenocarcinoma patients. A DeNovix DS-11 ultraviolet spectrophotometer was used to measure the concentration and purity of extracted RNA. Amplification refractory mutation systems (ARMS) and ABI 7500 fluorescence qPCR were used to detect ALK gene. Through statistical analysis, the CEM was compared with the CSM and CBM in RNA concentration, purity, and ALK gene detection results. RESULTS: The concentration of RNA extracted by using the CEM was significantly higher than that extracted by using the CBM and CSM (p < 0.001). The purity of RNA extracted by using the CEM was significantly higher than that by the other 2 methods (p = 0.011, p = 0.005). ALK gene testing with PCR was successful in all the samples using the CEM, but 2 cases by the CSM and 1 case by the CBM failed. CONCLUSIONS: Using the disposable membrane cell collector to process BPE of lung adenocarcinoma patients for RNA extraction and ALK gene detection is more effective and successful compared with the traditional methods, and it is suggested to be further applied and popularized in clinical practice.


Assuntos
Adenocarcinoma de Pulmão/genética , Quinase do Linfoma Anaplásico/genética , Biomarcadores Tumorais/genética , Separação Celular/instrumentação , Equipamentos Descartáveis , Fusão Gênica , Neoplasias Pulmonares/genética , Membranas Artificiais , Derrame Pleural Maligno/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Adenocarcinoma de Pulmão/enzimologia , Adenocarcinoma de Pulmão/patologia , Centrifugação , Humanos , Neoplasias Pulmonares/enzimologia , Neoplasias Pulmonares/patologia , Inclusão em Parafina , Derrame Pleural Maligno/sangue , Derrame Pleural Maligno/enzimologia , Derrame Pleural Maligno/patologia , Valor Preditivo dos Testes , Fixação de Tecidos
4.
Int J Clin Exp Pathol ; 13(8): 2106-2114, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32922607

RESUMO

OBJECTIVE: To explore the significance of a DNA quantitative analysis of lung cancer cells with different pathological types taken from bronchial brush specimens and its relationship with the clinicopathological features. METHODS: 903 bronchial brush cytological specimens taken in the Cytology Department of the Fourth Hospital of Hebei Medical University were collected from March 2017 to December 2019 and divided into three groups: the squamous carcinoma (SC) group, the adenocarcinoma (AC) group, and the small-cell carcinoma (SCC) group. The maximum DNA index (DI) value, the percentage of cells with a DI larger than 2.5, the mean DI, and the peaks of the aneuploid cells of the three groups were compared. A single factor analysis was used to analyze the relationship between the DNA content, aneuploidy, and the clinico pathological features of the patients who had surgery. RESULTS: The peaks of the aneuploid cells in the SC group, the AC group, and the SCC group had no significant differences (P=0.57). The maximum DI, the percentage of cells with a DI larger than 2.5, and the mean DI of the three groups showed statistically significant differences (P<0.001). The clinicopathological features of the AC patients and the SC patients, such as gender, age, tumor type, the maximum tumor diameter, clinical stage, vascular invasion, nerve invasion, pleural invasion, tracheal spread, and lymph node metastasis were not independent factors that influence the DNA content and have no statistical significance (P > 0.05). CONCLUSION: The reason why the DNA content of small-cell lung cancer is lower than SC and AC remains to be further studied.

5.
Int J Clin Exp Pathol ; 13(7): 1662-1668, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32782686

RESUMO

OBJECTIVE: The aim of this study was to use cerebrospinal fluid (CSF) cytology to give undiagnosed patients admitted to the hospital with severe neurological symptoms and without any anti-tumor treatment history a definitive diagnosis. Further, the aim was to explore the relationship between the frequency of gene mutations and mortality on the incidence of CSF metastasis in advanced non-small-cell lung cancer (NSCLC). MATERIALS AND METHODS: 30 patients diagnosed with NSCLC through CSF cytology were retrospectively analyzed. We analyzed 30 CSF metastasis patients and a control group of 20 advanced NSCLC patients without CSF metastasis. RESULTS: 30 patients were diagnosed with CSF metastasis using CSF cytology and immunocytopathology. The frequencies of EGFR mutations and ALK fusion in the CSF metastasis group were higher than they were in the non-CSF metastasis group (80% and 50% respectively, P<0.05). The incidence of CSF metastasis with gene mutations was higher than it was with wild-type genes (70.6% and 37.5%, P<0.05), OR 4.0 (95% CI 1.14~13.99). The median survival time of the CSF metastasis group was 4.8 months (95% CI 4.2~5.3). However, the median survival time in the non-CSF metastasis group was 9.2 months (95% CI 3.3~15.1). The mortality of the CSF metastasis group (n=13, 43.3%) was significantly higher than it was in the non-CSF metastasis group (n=6, 30%). CONCLUSIONS: CSF metastasis in NSCLC patients has a higher frequency of gene mutations and mortality. EGFR mutation and ALK fusion patients have a higher incidence of CSF metastasis. EGFR mutations and ALK fusion may promote CSF metastasis and may be a predictor of prognosis in NSCLC patients.

6.
Pathol Res Pract ; 216(8): 153036, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32703499

RESUMO

OBJECTIVE: To investigate the mutation status and clinical characteristics of multigene detection in advanced lung adenocarcinoma using cytological specimens. MATERIALS AND METHODS: 137 advanced lung adenocarcinoma patients with 10 driver genes detection in the Fourth Hospital Hebei Medical University from January 2019 to November 2019 was analysized. 137 cytological specimens including fine-needle aspiration specimens and maligant serous cavity effusion (pleural effusion, peritoneal and pericardial effusion). Ten driver mutations of EGFR, ALK, ROS1, BRAF, KRAS, NRAS, HER2, RET, PIK3CA and MET were detected by the amplification refractory mutation system (ARMS). Meanwhile, 90 of 137 patients were detected with biopsies for parallel gene detection. RESULTS: 78.10 % (107/137) of patients with advanced lung adenocarcinoma harbored at least one of 10 driver mutations. The three main mutations were EGFR (69.16 %, 74/137), ALK (6.57 %, 9/137)and ROS1 (3.65 %, 5/137) mutations. Besides, we found 6 cases including two concomitant mutations: EGFR Exon19 del/HER2 (1/137), EGFR Exon21 L858R/PIK3CA (2/137), EGFR Exon21 L858R/RET (1/137), and ALK/KRAS (2/137). Among 137 patients, women aged 64 or older were more likely to have the mutations (P < 0.05). Female patients (P = 0.003) older or equal to 64 years (P = 0.015) with non-smoking habbit (P = 0.027) were more detected with EGFR mutations, while ALK was more detectable in patients yonger than 64 years. Parallel analysis showed that rates of single EGFR, ALK, ROS1, RET, KRAS, NRAS, HER2, MET mutations and concomitant different mutations were not significantly different between cytological specimens and matched histological specimens. CONCLUSIONS: In the study, cytological specimens and biopsy samples have a very high coincidence rate of gene detection. EGFR, ALK and ROS1 mutations were the main driver mutations in patients with advanced lung adenocarcinoma.We speculate that EGFR and ALK are more prone to concomitant mutations respectively and the treatment of advanced lung adenocarcinoma patients with concomitant mutations deserves further study. The rate of KRAS, NRAS, BRAF, PIK3CA, RET and MET exon14 skipping mutation were low but may had a significant impact on the targeted therapy of patients with advanced lung adenocarcinoma.


Assuntos
Adenocarcinoma de Pulmão/genética , Citodiagnóstico/métodos , Análise Mutacional de DNA/métodos , Neoplasias Pulmonares/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Biópsia por Agulha Fina/métodos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Manejo de Espécimes
7.
J Cancer ; 11(8): 2265-2272, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32127953

RESUMO

Objectives: Lung adenocarcinomas with or without epidermal growth factor receptor (EGFR) mutations have shown different drug effects against EGFR inhibitors. But it is not very clear if EGFR mutation status affects the biological behavior of lung adenocarcinoma, because tumor gene regulation is very complicated and can be affected by many factors. We aimed to explore if EGFR mutation status is related with tumor malignant degree by investigating the relevance of EGFR mutation status with DNA content and aneuploid peaks of lung adenocarcinoma cells in pleural fluids without using EGFR-TKIs. Materials and Methods: 591 cases of lung adenocarcinoma patients in Hebei Tumor Hospital who had undergone EGFR gene detection and DNA quantitative analysis were collected from January 2012 to August 2018.They were divided into two groups: EGFR mutant group and non-mutant group. EGFR mutations were detected by Amplification Refractory Mutation System (ARMS) and ABI 7500 Fluorescence quantitative PCR with pleural effusions. DNA content and aneuploid peaks were detected by LD DNA image cytometry (DNA-ICM). Rank-sum test of SPSS 16 was used for statistical analysis. Results: The maximum DI, the mean DI of the first 20 cells greater than 5C, the percentage of cells greater than 5C and the number of cells greater than 9C of the first 20 cells in the mutant group were all higher than those in the non-mutant group, having statistical significance (p<0.001); the peaks of aneuploid cells in the mutant group occurred more often than those in the non-mutant group, having statistical significance (p<0.001). Conclusions: Our study has shown that advanced lung adenocarcinomas with EGFR-mutations had higher DI values, more aneuploid cells and more frequent aneuploid peaks compared with those without EGFR-mutations, suggesting that advanced lung adenocarcinomas with EGFR mutations are more aggressive than those without EGFR mutations.

8.
Int J Clin Exp Pathol ; 13(2): 132-141, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32211093

RESUMO

Non-small cell lung cancer (NSCLC) is the leading cause of cancer-related mortality all over the world, particularly in China. Metastasis is the main factor resulting in the poor prognosis of patients with NSCLC. CXCR4 and EGFR have been widely studied due to their critical role in tumor metastasis, but it remains more elusive then the relationship between CXCR4 and EGFR. Studies have demonstrated that many tumors have been found the existence of the "cross-talk" between EGFR and CXCR4 signaling pathways. In this context, we explored the relationship between EGFR and CXCR4 signaling pathways in lung cancer invasion and metastasis by both in vitro and in vivo experiments.

9.
Medicine (Baltimore) ; 98(22): e15843, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31145331

RESUMO

A clinically useful immune biomarker could potentially assist clinicians in their decision making. We stimulated T-cell proliferation to secret interferon gamma (IFN-γ) by phytohemagglutinin, and then measured the production of IFN-γ (mitogen value [M value]). We aimed to determine the relationship between the M value, clinical severity, and outcomes of diseases.In all, 484 patients admitted to intensive care units were enrolled in this retrospective study. The Acute Physiology and Chronic Health Evaluation II (APACHE II) scores were collected within the first 24 hours. M value, C-reaction protein (CRP), procalcitonin (PCT), erythrocyte sedimentation rate (ESR), and routine blood tests were analyzed and collected during the study.When APACHE II scores were greater than 15 and M values were less than 6, the hospital mortality rose in a straight line. There was an inverse correlation between APACHE II score and M value (rs = -0.212, P < .001). There was a positive correlation between M value and lymphocyte numbers (b' = 0.249, P < .001); however, there was an inverse correlation between M value and WBC (b' = -0.230, P < .001), and ESR (b' = -0.100, P = .029). Neurological diseases had the greatest influence on APACHE II scores (b' = 10.356, P < .001), whereas respiratory diseases had the greatest influence on M value (b' = 1.933, P < .001). Furthermore, in the respiratory system, severe pneumonia had a greater influence on M value. Taking the APACHE II score as the gold standard, the area under the curve of M was 0.632 (95% confidence interval [CI] 0.575-0.690, P < .001), PCT was 0.647 (95% CI 0.589-0.705, P < .001), CRP was 0.570 (95% CI 0.511-0.629, P = .022), and ESR was 0.553 (95% CI 0.494-0.612, P = .078). Divided by M value = 5, the positive predictive value of the M value is 37.22% (115/309) and negative predictive value is 75.43% (132/175).The results show that the M values, PCT, and CRP were better than ESR to predict the severity of diseases. The number and proportion of lymphocytes also affected the result of the M value. To a certain extent, the M value may be a clinically useful immune biomarker, which may help clinicians objectively evaluate the severity of diseases, especially in the respiratory system.


Assuntos
APACHE , Interferon gama/sangue , Mitógenos/administração & dosagem , Fito-Hemaglutininas/administração & dosagem , Doenças Respiratórias/sangue , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Área Sob a Curva , Biomarcadores/sangue , Sedimentação Sanguínea , Proteína C-Reativa/análise , Feminino , Humanos , Unidades de Terapia Intensiva , Contagem de Linfócitos , Masculino , Pessoa de Meia-Idade , Mitógenos/imunologia , Doenças do Sistema Nervoso/sangue , Fito-Hemaglutininas/imunologia , Pneumonia/sangue , Valor Preditivo dos Testes , Pró-Calcitonina/sangue , Estudos Retrospectivos , Adulto Jovem
10.
Cell Biol Int ; 38(10): 1155-62, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25044980

RESUMO

While Bcl-2 protein is involved in the regulation of apoptosis, recent research showed that Beclin1, described as the essential autophagy effector and haploinsufficient tumor suppressor, was originally isolated as a Bcl-2 interacting protein. Beclin1 interacts with Bcl-2 through a BH3 domain; nevertheless, the function of the anti-apoptotic gene, Bcl-2, in autophagy is not well understood. We explored the role of Bcl-2 in autophagy in human SGC-7901 cells in which Bcl-2 is overexpressed. Knockdown of Bcl-2 by small interfering RNA in human SGC-7901 cells downregulated Bcl-2 protein levels ∼82% and induced autophagy. Beclin1 protein, the first identified autophagy gene product, was induced by as much as 58%. Transmission electron microscopy and DNA fragmentation assay showed that autophagy was enhanced, but not apoptosis, in Bcl-2 siRNA treated cells. The results provide evidence that knockout the anti-apoptotic gene Bcl-2 induces autophagy in SGC-7901 cells and Bcl-2 specific siRNA may be used as a potential therapeutic strategy in gastric cancer cells that overexpress Bcl-2.


Assuntos
Proteínas Reguladoras de Apoptose/metabolismo , Autofagia , Regulação para Baixo , Proteínas de Membrana/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , RNA Interferente Pequeno/metabolismo , Proteína Beclina-1 , Linhagem Celular Tumoral , Fragmentação do DNA , Humanos , Proteínas Proto-Oncogênicas c-bcl-2/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-bcl-2/genética , Interferência de RNA
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