ABSTRACT
Objective To investigate the significance of abnormal gene in histologically normal mammary epithelial tissue for the early diagnosis of breast cancer. Methods Microarray technology was used to identify abnormal gene expres-sion and analyzed bioinformatics of normal mammary epithelial tissue in breast cancer patients and healthy normal control to establish a model for early diagnosis of breast cancer. The differentially expressed genes were screened by using signal path-way enrichment analysis. The accuracy (Ac), sensitivity (Sn) and specificity (Sp) were used to measure the prediction accura-cy of the different methods. Results The best prediction model was derived from the combination of differential genes en-riched from KEGG and BioCarta database. The number of differential expressed genes in three random created prediction models was reduced from 22 to 7, 14 to 3 and 18 to 4. However, the prediction accuracy was consistent with the model estab-lished from all of the differentially expressed genes, and the average accuracy of all models was 96.3%. Conclusion The prediction model can be simplified with the prediction accuracy unchanged, and thus facilitate the model apply to early diag-nosis and prevention of breast cancer.
ABSTRACT
Objective:To highlight the developmental process of 3D cell culture technology system, which is more suitable for isolating and identifying lung cancer stem-like cells than 2D cell culture technology system, and to explore the application of 3D cell cultures in the evaluation of proliferation, apoptosis, invasion, and drug resistance of lung cancer. Methods:Cells (104/well) from the human lung adenocarcinoma cell lines A549 and RPMI 1640 were cultured in complete medium containing 10%fetal bovine serum. Cell suspension was cultured in a BME basal medium. A growth curve was drawn after 7 d of culture. The stem-like cell was identified through a mammosphere culture, drug resistance and invasion assay, and flow cytometry. Data of A549 cells cultured in 3D and 2D tra-ditional cell culture technologies were compared. Results: Cells from the 3D cell culture had higher tumor formation rates [(20.75 ± 0.85) d vs. (60.25 ± 1.49) d, P<0.01)] and tumor sphere formation (28.50%± 1.17%vs. 8.67%± 0.80%, P<0.01) than those from the 2D cell culture. Moreover, cells from 3D cell culture were more invasive and resistant to therapy (58.17%± 2.19%vs. 41.70%±5.81%in 48 h, P<0.01;33.27%±5.76%vs. 27.30%±4.25%in 72 h, P<0.01). Phenotype experimental results demonstrated that the CD44 and CD326 cells were double-positive, whereas the CD24 cell was negative. Conclusion:The proportion of stem-like cells in A549 cell line after 3D cell culture significantly increased compared with 2D cell culture. The 3D cell culture can promote the proliferation of lung cancer stem cells.
ABSTRACT
Objective: This study aimed to examine the number of activated circulating endothelial cells (aCECs) in the peripheral blood of patients with non-small cell lung cancer (NSCLC), and investigate the relationship among aCECs, anti-angiogenic therapy, and prognosis of NSCLC patients. This study also aimed to identify novel predictive markers for anti-angiogenic therapy, and provide basic data and experimental basis for establishing an evaluation system for this therapy. Methods: A total of 142 NSCLC patients were randomly divided into the chemotherapy group (Group 1) and combined therapy group (i.e., chemotherapy plus endostatin, Group 2). The number of aCECs was measured using flow cytometry by detecting the expression status of CD105 and CD146 in the peripheral blood. The correlation between the changes in aCECs and efficacy of drug treatment was statistically analyzed using SPSS software. Results:The number of aCECs in Group 2 increased significantly at 8 and 29 d, two cycles, 50 and 71 d, and four cycles after treatment, respectively (P<0.05). In particular, aCECs amount in cases of progressive disease increased more significantly after combined therapy (P<0.05). A negative correlation was found between the treatment cycle and difference in aCECs amount before and after therapy (r=-0.970, P=0.001). A negative correlation was also observed between the difference in aCECs amount and time to tumor progression (TTP) (r=-0.351, P=0.039). Therefore, the difference in aCECs amount before and after therapy could serve as an important predictor for TTP in NSCLC patients. Conclusion:CD105 and CD146 reflected the activation status of endothelial cells, and responded to the drug treatment. Thus, CD105 and CD146 could act as ideal markers for aCECs. The number of aCECs increased during cancer progression, but significantly decreased after long-term treatment. Therefore, the change in aCECs amount may be a useful marker in predicting the efficacy of anti-angiogenic therapy.
ABSTRACT
Tissue factor pathway inhibitor-2 (TFPI-2), a member of the Kunitz-type family, is a broad-spectrum serine proteinase inhibitor. The expression of TFPI-2 is inversely related to increasing degree of malignancy, suggesting a role of TFPI-2 in the mainte-nance of tumor stability and inhibition of the growth of neoplasma. Aberrant methylation of TFPI-2 promoter cytosine-phosphorothio-ate-guanine (CpG) islands has been widely documented to be responsible for diminished expression of TFPI-2 mRNA and protein dur-ing cancer progression. TFPI-2 expression is significantly up-regulated by the ERK1/2 and JNK signaling pathways and modestly in-creased by VEGF, TNF-alpha, and fibroblast growth factor in time-and dose-dependent manners. TFPI-2 can maintain the stability of the tumor environment and inhibit invasiveness and growth of neoplasms. TFPI-2 has also been shown to regulate proliferation, apopto-sis, and vasculogenic mimicry of tumor cells, which may contribute significantly to tumor growth inhibition. Restoration of TFPI-2 ex-pression in tumor tissue inhibits tumor growth and metastasis, which creates a novel possibility of cancer patient treatment. This review focuses on the expression and the molecular regulation mechanisms of TFPI-2 in malignant tumors that control the functions of TFPI-2 in proliferation, apoptosis, and angiogenesis. Insight into these processes will improve our understanding of TFPI-2 and provide new ap-proaches for rational treatment strategies.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the expression of apoptotic regulator c-FLIP(L) in invasive breast carcinoma tissues, and to evaluate its correlation with molecular subtyping and clinical prognosis.</p><p><b>METHODS</b>Immunohistochemistry using EnVision staining for c-FLIP(L) was performed in 264 cases of invasive breast carcinomas and matched adjacent normal breast tissue samples from January 1996 to December 1999. ER, PR, HER2, Ki-67, CK5/6 and EGFR were evaluated by immunohistochemistry in order to classify the tumors into five molecular subtypes and the difference of c-FLIP(L) expression in these molecular subtypes was also analyzed. The influence of c-FLIP(L) expression on prognosis was evaluated by Kaplan-Meier curves and multi-factor Cox proportional risk model.</p><p><b>RESULTS</b>High expression of c-FLIP(L) was observed in 84.5% (223/264) of cases of invasive breast carcinomas which were significantly higher than the 45.1% (119/264) of cases in adjacent normal epithelium of breast (χ² = 89.78, P = 0.000). The expression of c-FLIP(L) in luminal B (HER2 positive) and basal-like breast cancers was 78.1% (25/32) and 46.2% (18/39), respectively, with significant difference (P < 0.05). Moreover, the expression of c-FLIP(L) in luminal B (HER2 positive) was higher than in luminal A cancers (P < 0.05), and the expression of c-FLIP(L) in HER2 positive cancers was higher than in basal-like cancers (P < 0.01). C-FLIP(L) showed deep yellow staining in node positive breast cancer with a high-expression rate of 93.1% (134/144); whereas the expression was sporadic and light yellow in node negative breast cancer with a lower high-expressed rate of 72.5% (87/120, P < 0.01). C-FLIP(L) expression had significant influence on disease-free survival time, with c-FLIP(L)-positive patients showing poor prognosis (P < 0.01). Multi-factor Cox proportional risk model analysis showed that expression of c-FLIP(L), lymph nodes status and molecular subtypes were independent prognostic factors for invasive breast carcinomas (P < 0.05).</p><p><b>CONCLUSIONS</b>C-FLIP(L) is highly expressed in invasive breast carcinomas, and its expression level is closely related to the molecular subtypes and clinical prognosis of breast cancer patients. Thus, c-FLIP(L) could be used as an important tumor marker for personalized cancer therapy and prognostic prediction.</p>
Subject(s)
Aged , Female , Humans , Biomarkers, Tumor , Metabolism , Breast , Metabolism , Breast Neoplasms , Classification , Metabolism , Mortality , CASP8 and FADD-Like Apoptosis Regulating Protein , Metabolism , Disease-Free Survival , Immunohistochemistry , Prognosis , Receptor, ErbB-2 , MetabolismABSTRACT
Purpose:To analyze the immunophenotype character of malignant lymphoma with bone marrow involvement by means of Flow Cytometry (CD45/SSC gate).Methods:Bone marrow of malignant lymphoma patients was detected by Flow Cytometer(CD45/SSC gate),bone marrow smear was simultaneously performed as control.Results:(1) Bone marrow of 34 malignant lymphoma patients was examined by Flow Cytometry.23 cases were detected to have bone marrow involvement.(2) Among these 23 cases,19 cases were non-Hodgkin's lymphoma(NHL),4 cases were Hodgkin's lymphoma(HL).The highest frequency antigen marker of B cell NHL was CD19 and CD20,T cell NHL was CD7,and HL was CD9.Conclusions:(1)Flow Cytometry(CD45/SSC gate) is a feasible and effective method to detect patients with bone marrow involvement.(2) The antigen marker of B cell NHL is CD19 and CD20,T cell NHL is CD7 and HL is CD9.
ABSTRACT
Objective: To explore the effects of various concentrations IFN-? on human oral epidermic cancer cell line KB and its multidrug resistant counterpart KBv200.Methods: The toxicity of IFN-? and/or VCR was assayed by MTT method; intracellular rhodamin123 concentration and p-glycoprotein expression were measured by flow cytometry; mdr-1 mRNA was assayed by RT-PCR. Results: High-dose IFN-?(10 000 IU/ml) had significant antiproliferative effects on KB/KBv200 cell lines. Low-dose IFN-? did not have any significant effect on growth of KB/KBv200 cells, but did increase the cytotoxic effect of VCR and the accumulation of Rhodamine123 in KBv200 cell line and had no effect on parent VCR-sensitive KB cell line. IFN-? down-regulated expression of P-gp and mdr-1mRNA . Conclusions: IFN-? has significant effect on the reversal of multidrug resistance of KBv200 cell line via a mechanism of P-gp and mdr1 mRNA down-regulation.
ABSTRACT
Objective:To study the direct effects of gradient concentration inteiferon-a(IFN-a) on lymphoma cells line Daudi and Jur-kat and a group of 15 cases refractory lymphoma. Methods: The effects of IFN-a on the growth of tumor cells were assayed by MTT, apoptosis were measured by flow cytometty, TUNEL and electron-microscope, 15 refractory lymphoma cases were treated by local injection of IFN-a combination chemotherapy.Results: Low-dose IFN-a did not have any significant effects on growth of Daudi and Jurkat cells, high-dose IFN-a (10 000U/ml) not only have significant anti-proliferative effects with dependence of time and dosage but also can induce apoptosis on both Daudi and Jurkat cells. 5 patients achieved a complete remission and 7 patients achieved a partial remission, the overall response rate was 80% .Conclusion:IFN-a( 10 000 U/ml) had significant anti-proliferative effects and inducing apoptosis on lymphoma cells.Local injection of IFN-a is one of effective therapy on refractory lymphoma.