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Objective@#To investigate the clinical characteristics, therapy modality and prognosis of primary breast diffuse large B-cell lymphoma(PB-DLBCL).@*Methods@#A total of 68 patients with PB-DLBCL treated in Tianjin Medical University Cancer Institute and Hospital were enrolled between January 1, 2004 and January 31, 2017. Clinicopathological data were retrospectively analyzed. 67 patients were female and only one male. The median age was 56 years old. 46 patients had Ann Arbor clinical stageⅠ~Ⅱ disease, and the other 22 were stage Ⅲ~Ⅳ. The patients with and without B symptom were 11 and 57, respectively. Kaplan-Meier method was used for univariate analysis to calculate the 5-year overall survival (OS) rate and 5-year progress-free survival (PFS) rate, compared using the log rank test. Cox regression analysis was used for multivariate analysis.@*Results@#The 1, 3, 5-year OS rate were 84.0%, 78.0% and 73.0%, and 1, 3, 5-year PFS rate were 80.0%, 71.0% and 51.0%, respectively. Univariate analysis indicated that eastern cooperative oncology group (ECOG) score, Ann Arbor clinical stage, international prognostic index (IPI) score, risk stratification, B symptom, β2-microglobulin(β2-MG) level, size of the tumor and cycles of chemotherapy were prognostic factors for OS (all P<0.05), and Ann Arbor clinical stage, IPI score, risk stratification and B symptom were prognostic factors for PFS (all P<0.05). Multivariate analysis indicated that Ann Arbor clinical stage was independent prognostic factor for OS(P=0.029) and B symptom was independent prognostic factor for PFS(P=0.028).@*Conclusions@#Prognosis of PB-DLBCL was relatively good. Ann Arbor clinical stage and B symptom were independent prognostic factors for OS and PFS, respectively.
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Objective: To construct an aromatase-overexpressing breast cancer cell model and observe the real-time apoptosis of breast cancer cells induced by the aromatase inhibitor, letrozole. Methods: The lentivirus-mediated gene transfection method was used to construct the MCF-7-VC3AI and ZR7530-VC3AI cell lines,which stably expressed the apoptotic fluorescent indicator protein VC3AI.Simultaneously,letrozole-induced apoptosis models of the MCF-7-VC3AI and ZR7530-VC3AI breast cancer cell lines were also constructed.Real-time quantitative PCR(qPCR)and Western blot methods were used to detect the mRNA and protein levels of aroma-tase in the cells.Cell proliferation ability was measured using MTT.The proliferation of cells in vitro under testosterone,estradiol,or letrozole combined with testosterone treatments were also observed.Results:qPCR results showed that the expression of the aroma-tase mRNA was significantly higher in both the MCF-7 and ZR7530 cell models when compared to the MCF-7-VC3AI and ZR7530-VC3AI cell models.Western blot results showed that the expression of the aromatase protein was significantly increased in both cell models. MTT assay results showed that the proliferation of a cell model could be promoted by testosterone and estrogen stimulation.Under 100 nmol/L testosterone,the proliferation rate of over-expressed aromatase MCF-7-VC3AI cells was about 1.2 times than that of the control group(P<0.01)and the proliferation rate of ZR7530-VC3AI cells was about 1.5 times than that of the control group(P<0.01). However,letrozole inhibited the proliferation induced by testosterone in a dose-dependent manner.Under the effect of letrozole at 10 μmol/L,the proliferation rate of over-expressed aromatase MCF7-VC3AI cells was 80% of the control group(P<0.05),while the prolifer-ation rate of over-expressed aromatase ZR7530-VC3AI cells was 68% of the control group(P<0.05).Conclusions:The successful estab-lishment of cell models that can detect letrozole-induced apoptosis provides an important foundation for further investigating the mechanism of letrozole.
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Objective To observe the clinical effect of lobaplatin combined with IL -2 in the treatment of malignant pleural effusion .Methods 42 patients with malignant pleural effusion in Datong Coal Group Co Cancer Hospital form 2014 to 2016 were enrolled as observation group,and treated with lobaplatin combined with IL -2. Another 45 patients with malignant pleural effusion in the same period were selected as control group ,and treated with lobaplatin.The efficacy and toxicity between the two groups were compared .The VEGF,CEA and TSGF were recorded in the two groups .The quality of life was evaluated by QLQ-C30 .Results The CR and PR rates in the observation group were 35.7％and 45.2％,respectively,which in the control group were 15.6％,22.2％,respectively,the differ-ences between the two groups were statistically significant (χ2 =4.672,5.178,all P<0.05).Both two groups showed different degrees of adverse reactions in treatment period,but there was no statistically significant difference in the incidence rate(P>0.05).The levels of VEGF,CEA and TSGF in the observation group were (194.6 ±25.4)ng/L, (42.4 ±9) ng/L and (81.3 ±10.3)U/mL,respectively,which were significantly lower than those in the control group[(195.0 ±24.1)ng/L,(43.1 ±9.5)ng/L,(80.9 ±9.4)U/mL](t =-0.075,-0.340,0.189,all P<0.05 ) .The improvement of dyspnea symptoms ,mood and overall health status in the observation group was better than those in the control group (t =23.326,-2.275,10.757,all P <0.05).Conclusion Lobaplatin combined with IL-2 pleural perfusion has significantly curative effect ,it can improve the quality of life and without increased toxicity in the treatment of malignant pleural effusion .
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Objective:To investigate the mechanism of histone deacetylase (HDAC) inhibitor in down-regulating the expression of HER-2 in breast cancer cells and to provide an innovative therapeutic option to overcome the disadvantages of anti-HER-2 therapy. Meth-ods:HER-2-positive breast cell lines were treated with HDAC inhibitors. The changes in the gene and protein levels of HER-2 were de-tected by qPCR and Western blot. MiRNA microarray was used to identify the HDAC inhibitors, whereas qPCR was used to verify the miRNA expression. Results:In vitro cell experiments confirmed that the HDAC inhibitors TSA and SAHA can down-regulate the expres-sion of HER-2 in breast cancer cell lines. TSA can down-regulate the expression of HER-2 gene in BT474 and decrease the concentra-tions of 100 nmol by 10.7%and 200 nmol by 38.9%(P<0.05). TSA had no effect on the primary cells. The expression of HER-2 gene of BT474 was down-regulated by 93.9%(P<0.05) in the 5μmol/L group but not in the 1μmol/L group. SAHA significantly affected the pri-mary cells at a concentration of 1μmol/L and reduced the cells at 87.1%at a concentration of 5μmol/L. Seven miRNAs were identified from the miRNA microarray. MiR-762 was used as a basis to identify the changes in miRNA. The miRNA sputum identified by miRNA microarray and qPCR may be associated with the down-regulation of HER-2 by HDAC inhibitors. Conclusion: HDAC inhibitors may down-regulate the expression of HER-2 in breast cancer cells by changing some miRNAs.
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Objective To investigate the effects and possible mechanisms of Mfn-2 gene overexpression on photodynamic therapy (PDT) sensitivity of T47D cells in human breast cancer.Methods pEGFP and pEGFP-Mfn-2 were transfected into human breast cancer T47D cells,and then the mRNA and protein expression of Mfn-2 gene in T47D cells were detected by real-time PCR (RT-PCR) and Western blotting in pEGFP group and EGFP-Mfn-2 group,respectively.After pEGFP and pEGFP-Mfn-2 were transfected into human breast cancer T47D cells for 48 hours,methyl thiazolil tetracolium (MTT) assay was used to measure the PDT sensitivity of T47D cells in human breast cancer in pEGFP group and pEGFP-Mfn-2 group.pEGFP + PDT group and pEGFP-Mfn-2 + PDT group were obtained by PDT irradiating pEGFP and pEGFP-Mfn-2 which were transfected into human breast cancer T47D cells.Cell apoptosis and mitochondrial membrane potential of T47D cells were assayed by flow cytometry in pEGFP + PDT group and pEGFP-Mfn-2 + PDT group.Laser scanning confocal fluorescence microscope was applied to observe the morphological ultrastructure of mitochondria in pEGFP group,pEGFP-Mfn-2 group,pEGFP + PDT group,and pEGFP-Mfn-2 + PDT group.Results RT-PCR showed that after transfecting T47D cells with pEGFP,the expression of Mfn-2 mRNA was 1.01 ±0.12.After transfecting T47D cells with pEGFP-Mfn-2,the expression of Mfn-2 mRNA was 1 067.00 ±41.72.There was statistical significance (t =67.541,P ＜ 0.001).Western blotting revealed that compared with the pEGFP group,the pEGFP-Mfn-2 transfection group had higher expression of Mfn-2 gene in T47D cells.MTT assay showed that pEGFP-Mfn-2 transfection significantly enhanced the PDT sensitivity of T47D cells in hunan breast cancer compared with the pEGFP + PDT group.When the concentration of methylene blue was 5.00 μmol/ml,the survival rate of the pEGFP +PDT group and pEGFP-Mfn-2 + PDT group were (59.96％ ± 1.21％) vs.(46.50％ ± 1.72％),with significant difference (t =34.403,P ＜ 0.001).Flow cytometry assay showed that the cell apoptosis rates in pEGFP-Mfn-2 + PDT group was markedly higher compared with the pEGFP + PDT group [(81.21 ± 2.13)％ vs.(68.82 ±2.64)％,P=0.024],with statistical significance.Also,the mitochondrial membrane potential was obviously lower in pEGFP-Mfn-2 +PDT group compared with the pEGFP +PDT group [(1.37 ±0.12)％ vs.(23.33 ± 1.86)％,P＜0.001],with statistical significance.Laser scanning confocal fluorescence microscope showed that cells in pEGFP group showed network structure,both pEGFP-Mfn-2 group and pEGFP + PDT group could cause mitochondrial fusion,and the pEGFP-Mfn-2 + PDT group could induce mitochondrial disintegration and lose its normal morphology completely.Conclusion Mfn-2 may enhance the PDT sensitivity of T47D cells in human breast cancer,which is possibly related with the normal morphology alteration of mitochondria and the inducement of mitochondrial apoptosis.
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<p><b>OBJECTIVE</b>To elucidate the impact of Hes1 on the proliferation and apoptosis of acute myeloid leukemia (AML) cells.</p><p><b>METHODS</b>The expression levels of Hes1 and p21 in AML patient samples and myeloid leukemia cell lines were analyzed by real-time PCR. Hes1 was up-regulated by retrovirus transfection in AML cell lines and the proliferation capacity were assayed by MTT, cell cycle by Hoechst/PY, apoptosis by AnnexinV.</p><p><b>RESULTS</b>The expression of Hes1 in primary AML cells and HL-60, U937, KG1a cell lines were 0.67 ± 0.24, 0.59 ± 0.43, 0.42 ± 0.03, and 0.32 ± 0.26, respectively, and p21 were 0.54 ± 0.01, 0.44 ± 0.12, 0.36 ± 0.12, and 0.59 ± 0.43, respectively. Hes1 expression levels after transduction in HL-60, U937, KG1a were 4.9 ± 0.2, 5.2 ± 0.4, 5.8 ± 0.5, respectively. Induced activation of Hes1 led to AML cells growth arrest and apoptosis, which was associated with an enhanced p21 expression. Besides, activated Hes1 led to AML cells growth inhibition in vivo.</p><p><b>CONCLUSION</b>Hes1 could mediate growth arrest and apoptosis in AML cells, which may be a novel target for AML.</p>
Subject(s)
Humans , Apoptosis , Basic Helix-Loop-Helix Transcription Factors , Cell Cycle , Cell Line, Tumor , Homeodomain Proteins , Leukemia, Myeloid, Acute , Transcription Factor HES-1 , Up-RegulationABSTRACT
Objective:This study aimed to observe the synergistic effect of a new tumor vaccine combined with metronomic che-motherapy in vivo on breast cancer. This study was also conducted to investigate the mechanism of this combination. Methods:Balb/c mice inoculated with 4T1 mouse breast cancer cell were used as tumor models. High-mobility group nucleosome-binding protein 1 (HMGN1) gene was used to transfect 4T1 cell lines as cancer vaccines. After 4T1 cell was inoculated, the mice were randomized into four groups:normal saline (NS);metronomic gemcitabine (GEM) alone;cancer vaccine alone;and combination therapy group. Tumor growth and potential toxicities of these regimens were observed. The Foxp3 expression of regulatory T cells (Tregs) was detected by western blot and immunohistochemical staining. The microvessel density (MVD) of the tumor was also detected by immunohistochemi-cal staining. Results:The tumor volume of the mice was significantly lower in the combination group than in the MET group or cancer vaccine group (P<0.05). This result exhibited a higher significant difference than the tumor volume of the mice in the NS group (P<0.01). Foxp3 expression was significantly lower in the mice treated with GEM (combination or MET group). MVD was significantly lower in these two groups than in the cancer vaccine group or NS group (P<0.05). Furthermore, adverse reactions slightly occurred in each group. Conclusion: The combination of cancer vaccines and metronomic GEM is a very active and well-tolerated regimen for breast cancer in mice.
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Objective:To determine the effect of Hes1 on bone marrow CD34+cells in acute myeloid leukemia (AML). Meth-ods:Bone marrow mononuclear cells were isolated by using Ficoll. Then, the proportion and cell cycle of CD34+cells were analyzed by using fluorescence-activated cell sorting (FACS). CD34+cells were cultured in vitro for colony-forming cells (CFC). The expression of Hes1 in CD34+cells was evaluated by using real-time polymerase chain reaction. After upregulating the expression of Hes1 in CD34+cells, the cell cycle was analyzed through FACS, and the colony formation of CD34+Hes1+cells was analyzed by CFC. Results:The ra-tio of CD34+cells in the bone marrow was lower in the AML group than in the control group. In addition, more CD34+cells underwent quiescence in the AML group than in the control group. In vitro assay showed that the colony formation of CD34+cells was lower in the AML group than in the control group. The expression of Hes1 was higher in the CD34+cells from the AML patients than that in the CD34+ cells from normal donors. After Hes1 transduction, more CD34+ cells underwent quiescence and showed weak proliferation. Conclusion:The proportion of CD34+cells in the bone marrow was lower in AML patients than in normal donors. A large proportion of CD34+cells underwent quiescence, which was related to Hes1, in AML patients.
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Objective To construct the recombinant RIP3 over-expressed plasmids and transfect them in breast cancer MCF7 cells, and identify the expression and localization of fusion protein, as well as its effect on the death way of MCF7 cells. Methods The expression levels of RIP3 mRNA in four breast cancer cell lines and normal mammary epithelial cells were detected by reverse transcription polymerase chain reaction (RT-PCR). The RIP 3 coding sequence was amplified by polymerase chain reaction and subcloned into mCherry vector to construct recombinant plasmids. The plasmids were transfected into MCF7 cells by lentivirus after DNA sequencing, then screened by basticidin (4 mg/L) for 1 week. The efficiencies of RIP3 expression were validated by Western blotting assay. The death way of mCherry-RIP3-MCF7 cells was observed under the treatment of TNF-αand Z-VAD-FMK. Results The lowest expression of RIP3 mRNA was found in MCF7 cells. The sequencing results validated the well recombinant plasmids. The expression of mCherry-RIP3 fusion pro-tein with a molecular weight of 85 ku was detected by Western blot assay. The mCherry-RIP3 expression enhanced the sensi-tivity of MCF7 cells to TNF-αand Z-VAD-FMK induced cell death. Conclusion The recombinant RIP3 over-expressed plasmids were successfully constructed, and the stable MCF7 cells with ectopic RIP3 transfection were obtained. The mCher-ry-RIP3 fusion protein was expressed in the cytoplasm and was conformed to mediate TNF-αinduced necroptosis.
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Objective:To analyze the prognostic factors in patients with metastatic breast cancer (MBC). Methods:A total of 205 patients with pretreated MBC were included in this study. These patients were admitted to the Tianjin Medical University Cancer Insti-tute&Hospital and had undergone radical surgery of breast cancer between January 2008 and December 2010. The clinicopathologic information of the patients was collected in this retrospective analysis. Results: The median overall survival of the patients was 32 months (1 month to 132 months). Luminal A, Luminal B, HER-2 overexpression, and triple-negative patients had a median overall sur-vival of 36 months (4 months to 132 months), 32 months (7 months to 122 months), 29 months (1 month to 85 months), and 24 months (1 month to 98 months), respectively. Univariate analysis showed that lymph node metastases, clinical stage, molecular type, visceral disease, first multiple metastatic sites, and shorter metastasis-free interval were significantly associated with poor outcomes. In multivar-iate analysis, lymph node metastases, clinical stage, molecular type, visceral metastasis, and the number of first metastatic sites were significant predictors of patient survival. Conclusion:Lymph node metastasis, clinical stage, triple-negative breast cancer, and visceral metastasis were used as independent poor prognostic indicators for survival in patients. Results of this study may assist physicians in evaluating the survival potential and determining the appropriate therapeutic strategy for MBC patients.
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Familial non-medullary thyroid carcinoma (FNMTC) is de fi ned as the presence of two or more affected fi rst-degree relatives with non-medullary thyroid cancers without other known familial syndromes. FNMTC is one of the most inheritable forms of all cancers, with a high risk of a first-degree relative developing the disease. Compared with sporadic non-medullary thyroid carcinoma (NMTC), FNMTC presents at a younger age and is associated with a higher incidence of multifocal disease and metastasis. This in-creased aggressiveness has been hypothesized to translate into higher recurrence rates and decreased survival of patients with FNMTC. The genes involved in the pathogenesis of FNMTC are yet to be elucidated, although some recent studies identified several predisposi-tion loci with a high degree of genetic heterogeneity. Since 2005, next-generation sequencing (NGS) technologies have been developing as rapid, high-throughput, and cost-effective approaches to fulfill medical sciences and research demands. With the use of NGS, the un-derlying causative genes can be directly distinguished via systematic filtering, through which the identified gene variants are verified for novelty and functionality.
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Antitumor effects of erythromycin and the related mechanism were investigated in the present study. Neuroblastoma cells (SH-SY5Y) were exposed to erythromycin at different concentrations for different durations. Cell proliferation was measured by cell counting, and cell viability was examined by MTT assay. Cell cycle phase distribution and the cytosolic calcium level were detected by flow cytometry. Mitochondrial membrane potential was measured by the JC-1 probe staining and fluorescent microscopy. The expression of an oncogene (c-Myc) and a tumor suppressor [p21 (WAF1/Cip1)] proteins was analyzed by using Western blotting. Erythromycin could inhibit the proliferation of SH-SY5Y cells in a concentration- and time-dependent manner. The cell cycle was arrested at S phase. Mitochondrial membrane potential collapsed and the cytosolic calcium was overloaded in SH-SY5Y cells when treated with erythromycin. The expression of c-Myc protein was down-regulated, while that of p21 (WAF1/Cip1) protein was up-regulated. It was concluded that erythromycin could restrain the proliferation of SH-SY5Y cells. The antitumor mechanism of erythromycin might involve regulating the expression of c-Myc and p21 (WAF1/Cip1) proteins.
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Objective To investigate clinical biologic characteristics and factors that impact prognosis of patients with follicular thyroid carcinoma(FTC).Method Clinical data of 105 FTC patients treated surgically in the Department of Head& Neck,Tianjin Cancer Hospital from 1970 to 1990 was analyzed retrospectively.Results The overall 5-year,10-year and 15-year survival rates of these patients were 85.3％,76.7％ and 72.9％ respectively.The overall 15-year survival rates of patients ≥45 years and those ＜ 45 years were 45.9％,89.8％ respectively.The overall 15-year survival rates of patients with unilateral and bilateral carcinoma was 50.0％,76.2％ respectively.The overall 15-year survival rates of patients with neck lymph node metastasis and without were 54.2％,79.2％ respectively.The overall 15-year survival rates of patients at stage Ⅰ,stage Ⅱ,stageⅢ and stageⅣ were 89.3％,70.0％,45.5％ and 35.3％respectively.During the follow-up period,11 patients were diagnosed with distant metastasis from 1 year to 33 years after surgical treatment and 9 died of cancer within 5 years after diagnosis.Seventeen patients had local recurrence from 3 months to 34 years after surgery and 10 of these patients died of local recurrence.Conclusions The factors influencing prognosis of patients with FTC were age,clinical stage,bilateral carcinoma and neck lymph node metastasis,therefore early treatment and close following-up are essential to improve the prognosis of patients with FTC.