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Breast adenomyoepitheliomas are composed of a biphasic proliferation of myoepithelial cells around small epithelial-lined spaces. Due to the rarity of adenomyoepitheliomas, the molecular data describing them are limited. Adenomyoepitheliomas are considered to be benign or have low malignant potential, and be prone to local recurrence. Malignant transformation has been associated with homozygous deletion of CDKN2A or somatic mutations in TERT, but remains unexplained in many cases. Here, we describe a case of carcinomatous transformation of both epithelial and myoepithelial cells in an estrogen receptor-negative adenomyoepithelioma caused by amplification of MYC. Break-apart fluorescence in situ hybridization revealed an increase in the MYC gene copy number (3–4 copies/cell in 37%, > 4 copies/cell in 40%). Deregulation of MYC is responsible for uncontrolled proliferation and cellular immortalization in basal-like breast cancers. Our case demonstrates that genomic instability events associated with gene amplification may be involved in the carcinogenesis of malignant adenomyoepitheliomas.
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Adenomyoepithelioma , Breast Neoplasms , Breast , Carcinogenesis , Estrogens , Fluorescence , Gene Amplification , Genes, myc , Genomic Instability , In Situ Hybridization , In Situ Hybridization, Fluorescence , RecurrenceABSTRACT
Objective@#To observe the effect of different expression levels of USP28 on the radiosensitivity of ECA109 cells by gene transfection method, aiming to provide theoretical basis for comprehensive treatment of esophageal cancer.@*Methods@#The expression levels of USP28 and c-Myc in the esophageal epithelial cells Het-1A, ECA109 and ECA109R were quantitatively measured by qRT-PCR. The specific siRNA sequences were designed according to the USP28 and c-Myc genes. The pcDNA-USP28 and pcDNA-c-Myc plasmids were constructed. The esophageal cancer cell ECA109 was transfected with Lipofectamine 2000 to observe the transfection effect and related protein expression. ECA109 and ECA109R cells were exposed to 6 Gy X-ray radiation. The cell apoptosis in each group was detected by flow cytometry. The radiosensitivity was evaluated by clone formation assay.@*Results@#The expression levels of USP28 and c-Myc in ECA109 were significantly higher than those in Het-1A (both P<0.05), and the expression levels of USP28 and c-Myc in ECA109R were remarkably higher than those in ECA109(both P<0.05). The pcDNA-USP28 and pcDNA-c-Myc recombinant plasmids were successfully constructed. Compared with the negative control group, the expression of USP28 at the protein and mRNA levels in the si-USP28 group was significantly down-regulated, whereas those in the pcDNA-USP28 group were remarkably up-regulated. Similar results were obtained in terms of c-Myc. Compared with the control group, the expression level of c-Myc protein was significantly up-regulated in the pcDNA-USP28 group, whereas considerably down-regulated in the si-USP28 group. After 6 Gy irradiation, the apoptosis rate and radiosensitivity of ECA109 cells were significantly declined. The apoptosis rate and radiosensitivity of ECA109R cells were increased in the si-USP28 group.@*Conclusions@#The expression of USP28 protein is closely correlated with the radiosensitivity of esophageal cancer cells. The underlying mechanism may be related to the regulation of c-Myc expression by USP28.
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Objective To observe the effect of different expression levels of USP28 on the radiosensitivity of ECA109 cells by gene transfection method,aiming to provide theoretical basis for comprehensive treatment of esophageal cancer.Methods The expression levels of USP28 and c-Myc in the esophageal epithelial cells Het-1A,ECA109 and ECA109R were quantitatively measured by qRT-PCR.The specific siRNA sequences were designed according to the USP28 and c-Myc genes.The pcDNA-USP28 and pcDNA-c-Myc plasmids were constructed.The esophageal cancer cell ECA109 was transfected with Lipofectamine 2000 to observe the transfection effect and related protein expression.ECA109 and ECA109R cells were exposed to 6 Gy X-ray radiation.The cell apoptosis in each group was detected by flow cytometry.The radiosensitivity was evaluated by clone formation assay.Results The expression levels of USP28 and c-Myc in ECA109 were significantly higher than those in Het-1A (both P＜0.05),and the expression levels of USP28 and c-Myc in ECA109R were remarkably higher than those in ECA109(both P＜0.05).The pcDNA-USP28 and pcDNA-c-Myc recombinant plasmids were successfully constructed.Compared with the negative control group,the expression of USP28 at the protein and mRNA levels in the si-USP28 group was significantly down-regulated,whereas those in the pcDNA-USP28 group were remarkably up-regulated.Similar results were obtained in terms of c-Myc.Compared with the control group,the expression level of c-Myc protein was significantly up-regulated in the pcDNA-USP28 group,whereas considerably down-regulated in the si-USP28 group.After 6 Gy irradiation,the apoptosis rate and radiosensitivity of ECA109 cells were significantly declined.The apoptosis rate and radiosensitivity of ECA109R cells were increased in the si-USP28 group.Conclusions The expression of USP28 protein is closely correlated with the radiosensitivity of esophageal cancer cells.The underlying mechanism may be related to the regulation of c-Myc expression by USP28.
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AIM:To observe the effect of antisense locked nucleic acid (anti-LNA) blocking the translation initiation region of c-myc exon 2 on the viability and apoptosis of hepatocellular carcinoma cells.METHODS:The antiLNA that was complementary to the translation initiation region of c-myc exon 2 was designed,synthesized,and introduced into the HepG2 cells by cationic liposome-mediated transfection.The mRNA and protein levels of c-Myc in the cells were determined by RT-PCR and Western blot.The change of cell apoptosis was analyzed by flow cytometry,and the toxicity of anti-LNA to the cells was detected by MTT assay.RESULTS:Five days after transfection,the mRNA level of c-Myc in anti-LNA group was 0.335 ±0.016,and the protein level was 0.448 ± 0.037,significantly lower than those in control group (both P ＜ 0.05).The ratio of apoptotic cells in anti-LNA group was 32％ ±-6％,which was higher than that in control group (P ＜ 0.05).CONCLUSION:Antisense locked nucleic acid targeting at the translation initiation region of cmyc exon 2 shows strong inhibitory effects on the apoptosis of hepatocellular carcinoma cells.
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AIM:To observe the effect of antisense locked nucleic acid (anti-LNA) blocking the translation initiation region of c-myc exon 2 on the viability and apoptosis of hepatocellular carcinoma cells.METHODS:The antiLNA that was complementary to the translation initiation region of c-myc exon 2 was designed,synthesized,and introduced into the HepG2 cells by cationic liposome-mediated transfection.The mRNA and protein levels of c-Myc in the cells were determined by RT-PCR and Western blot.The change of cell apoptosis was analyzed by flow cytometry,and the toxicity of anti-LNA to the cells was detected by MTT assay.RESULTS:Five days after transfection,the mRNA level of c-Myc in anti-LNA group was 0.335 ±0.016,and the protein level was 0.448 ± 0.037,significantly lower than those in control group (both P ＜ 0.05).The ratio of apoptotic cells in anti-LNA group was 32％ ±-6％,which was higher than that in control group (P ＜ 0.05).CONCLUSION:Antisense locked nucleic acid targeting at the translation initiation region of cmyc exon 2 shows strong inhibitory effects on the apoptosis of hepatocellular carcinoma cells.
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Low frequency electromagnetic fields (LF-EMFs) which can be generated from homes and workplaces appliances can lead to alteration in oncogenes causing cancer diseases such as leukemia, nervous system tumors, lymphoma and breast cancer. To investigate the effect of LF-EMFs on c-myc oncogene expression level, primary cell culture of human peripheral blood mononuclear cells (PBMCs) were exposed to AC (50 Hz) electromagnetic flux density: 0.37mT, 0.82mT, 1.22mT, 1.68mT, 2.1mT, 2.47mT, 2.85mT, 3.33mT, 3.72mT, 3.92mT, 4.32mT and 4.67mT using exposure unit. After four days of exposure, when initial changes in cell viability, morphology and count between exposed and unexposed cells were noted, total RNA was extracted to evaluate the expression level of c-myc oncogene by quantitative real time PCR (qRT-PCR). The results showed that c-myc oncogene expression level began to increase gradually from value 1.69% at 0.82mT reaching to the maximum expression level value 4.65% at 4.67mT. This result refers to an increasing in LF-EMFs exposure offset by an increasing in c-myc oncogene expression level, affecting different cellular functions. This study indicates that LF-EMF is environmental pollution effects on the expression level of oncogenes which increase the risk of human cancer diseases.
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Objective:To investigate the clinical significance of Bcl-6, c-myc gene abnormalities in Xinjiang Uygur and Han dif-fused large B-cell lymphoma (DLBCL) subtypes. Methods:Bcl-6, c-myc gene was detected by fluorescence in situ hybridization in 233 patients with DLBCL . A relationship was observed among Bcl-6, c-myc gene translocation, and clinical data in DLBCL patients. In addition, a difference was observed among Bcl-6, c-myc gene translocation, and different ethnic groups in different subtypes of DLB-CL. Results:Among the 233 patients, 51 cases (21.89%) had rearranged Bcl 6 gene, and 39 cases (16.74%) had rearranged c-myc gene. Bcl-6 gene translocation and expression was related with age, gender, disease location, clinical stage, and LDH levels (P>0.05), but was not related with nationality , international prognostic index score, extranodal involvement, B symptoms, DLBCL subtypes, and recent efficacy (P0.05), but was not related with nationality, IPI score, extranodal involvement, B symptoms, and recent effica-cy (P0.05). By con-trast, in the Uygur and Han non-GCB groups, Bcl-6, c-myc gene translocation showed significant difference (P>0.05). Conclusion:Bcl-6, C-myc gene translocation was related with age, gender, disease location, clinical stage, and LDH levels. Bcl-6 gene translocation was also correlated with different subtypes of DLBCL.
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Objective To study the effect of p38MAPK on the activity and c-myc protein expression in rat acetaldehyde-induced hepatic stellate cell(HSC),and to investigate the alcoholic liver fibrosis related mechanism.Methods The different concentrations of SB203580 as the p38 specific blocker was adopted to conduct the intervention on rat acetaldehyde-induced HSC.The cellular mor-phological change was observed by the microscope.The cell proliferation was detected by MTT,the cell cycle was analyzed by flow cytometry(FCM),and the expression of c-myc protein was examined by the SABC method.Results (1)after acetaldehyde stimula-tion,HSC was increased in size and proliferated rapidly,but with the added SB203580 concentration increase,the cellular prolifera-tion was slowed down,the cells size was diminished and the deformed cells were increased.(2)The proliferation of acetaldehyde-in-duced HSC was inhibited by different doses of SB203580,and the higher concentration has the more significant inhibiting effect.(3) With the SB203580 concentration increase,the cells at the phase G0 and G1 were increased,while the cells at the phase S were de-creased,at the same time the expression positive rate of c-myc protein was decreased.Conclusion Blocking p38MAPK pathway ac-tivity could inhibit the proliferation of acetaldehyde-induced HSC,which may be related to the down-regulation of C-myc protein ex-pression and blocking the DNA synthesis in cells entering from G0/G1 phase to S phase.
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Objective: To analyze the diagnosis and therapy for double-hit lymphoma (DHL) and triple-hit lymphoma (THL). Methods:This study involves three patients with follicular lymphoma (FL) that transformed into DHL or THL. These patients were ad-mitted to our hospital between January 2011 and December 2012. All patients were diagnosed by immunohistochemistry and fluores-cence in situ hybridization (FISH). Results:One FL patient transformed into THL and died after 3 months. The other two FL patients who transformed into DHL and who received R-CHOP and R-ESHAP regimens still failed to achieve complete remission. Conclusion:DHL is a rare type of lymphoma that usually involves the bone marrow and central nervous system. This condition is highly resistant to intensive chemotherapy. Part of the DHL cases result from FL. FISH is important for diagnosis. However, a standard treatment for this type of lymphoma remains lacking.
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Introdução: A hipótese de que os efeitos do alumínio em células humanas podem ter implicações clínicas tem sido levantada há algum tempo, especialmente no que concerne ao câncer de mama. As evidências laboratoriais mostrando altos níveis de alumínio nos tecidos da mama e os efeitos biológicos conhecidos sobre esse metal não são suficientes para estabelecer uma relação causal entre a exposição ao alumínio e o risco aumentando para o desenvolvimento do câncer de mama. O objetivo deste estudo foi estabelecer a concentração de alumínio nas áreas centrais e periféricas de tumores de mama, assim como na área glandular normal da mama e correlacionar esses achados com a instabilidade dos genes ERBB2, C-MYC e CCND1 e a aneuploidia dos cromossomos que contêm estes genes. Métodos: Para este estudo foram incluídas 176 mulheres com diagnóstico de carcinoma invasor de mama, com tumores maiores de 1cm3, sem quimioterapia neoadjuvante, operadas enter 2008 e 2010 no Hospital da Mulher Prof. Dr. José Aristodemo Pinotti - Centro de Atenção Integral à Saúde da Mulher (CAISM) - UNICAMP. Para a análise da concentração de alumínio intracelular, amostras de 150 pacientes foram consideradas viáveis; para a análise da instabilidade genômica em função da concentração de alumínio, 118 amostras foram consideradas viáveis, definindo o espaço amostral de cada um dos artigos apresentados. As amostras das áreas centrais e periféricas dos tumores de mama e das áreas glandulares normais da mama foram obtidas. A quantificação do alumínio contido nos tecidos da mama foi feita através da técnica de Espectrometria de Absorção Atômica em Forno de Grafite (GFAAS). Uma lâmina de Tissue Microarray (TMA), contendo as amostras de tumor e tecido normal foi utilizado para a realização da técnica de FISH para acessar o status dos genes ERBB2, C-MYC e CCND1 e dos centrômeros dos seus respectivos cromossomos 17, 8 e 11. Os dados clínico-patológicos foram obtidos dos prontuários de pacientes...
Introduction: It has long been hypothesized if the effects of aluminum on human cells may have clinical implications, especially regarding to breast cancer. The current laboratorial evidence showing higher levels of aluminum in breast tissues and the known biological effects of this metal, are not sufficient to establish a causal relationship between aluminum exposure and increased risk of developing breast cancer. The objective of this study was to establish the aluminum concentration in the central and peripheral areas of breast tumors as well as in normal glandular area of the breast and to correlate these findings with the instability of ERBB2, C-MYC and CCND1, and aneuploidy of chromosomes harboring these genes. Methods: This study included 176 women diagnosed with invasive breast carcinoma with tumors larger than 1cm3 without neoadjuvant chemotherapy, operated between 2008 and 2010 at the Women's Hospital Professor. Dr. José Aristodemo Pinotti - Centro de Atenção Integral à Saúde da Mulher (CAISM) - UNICAMP. To analyze the intracellular concentration of aluminum, samples from 150 patients were considered viable; for the analysis of genomic instability as a function of the concentration of aluminum, 118 samples were considered viable. These figures define the sample of each of the two articles that this PhD thesis comprises. Evaluation of tissue aluminum content was carried out using Graphite Furnace Atomic Absorption Spectrometry (GFAAS). A TMA slide containing the tumor and normal samples was used in FISH assays to assess ERBB2, C-MYC and CCND1 and the respective chromosomes 17, 8 and 11 centromeres status. Clinicopathological data were obtained from patients' records. Results: The average aluminum content found in breast was 1.88 mg/kg in the central tumor areas, 2.10 mg/ kg in the peripheral tumor areas and 1.68 mg/ kg in the normal tissue areas...
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Humans , Female , Aluminum/adverse effects , Breast Neoplasms , Genomic Instability , Cyclin D1 , Genes, mycABSTRACT
Objective To investigate aberrations of bcl-6,p53,c-myc genes in diffuse large B-cell lymphoma (DLBCL) and its clinical significance.Methods Interphase fluorescence in situ hybridization (I-FISH) was detected in 59 DLBCL patients in vivo tissue bcl-6,p53 protein,c-myc gene status.The patients were treated with CHOP or R-CHOP chemotheralpy,and the survival rates and treatment efficiency were compared.Results The p53 deletion was detected in 18 of the 59 cases (30.5 ％),bcl-6 rearrangement in 11 cases (18.6 ％),5 cases with c-myc rearrangement (8.5 ％).In the aspects of remission rate,p53 deletion positive group contained less advantage than negative ones (33.3 ％ vs 75.6 ％,x2 =9.560,P =0.002).The prognosis of bcl-6 gene rearrangement positive group different from negative group,but the difference was not statistically significant (OS,P =0.107; PFS,P =0.094),p53 deletion positive patients was in significantly worse prognosis than the negative group (OS,P =0.031; PFS,P =0.028),c-myc rearrangement positive group difference in gene rearrangement negative group,but the difference was not statistically significant (OS,P =0.163; PFS,P =0.167).In the CHOP group,prognosis of p53 deletion,c-myc rearrangement positive group were significantly worse than the negative group,the difference was statistically significant (P ＜ 0.05).In R-CHOP group,the prognostic significance of bcl-6 gene rearrangement positive group were worse (OS,P =0.003; PFS,P =0.007).Conclusion DLBCL patients with bcl-6,p53,c-myc genes aberrations are related with poor prognosis,and they can be used as prognostic factors for predicting DLBCL and guiding therapy.
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Objective To investigate how SDF-1α and c-MYC protein regulates TACRl-Tr expression. Methods c-myc shRNA vector was constructed, small interfering RNA was employed for silencing c-myc gene in MCF-7 breast cancer cell. SDF-1α neutralized antibody was used in c-myc+ cell group and c-myc- cell group, while other c-myc+ cell group and c-myc- cells group were cultured under normal condition. The mRNA level of TACRl-Tr was determined by real-time PCR. Results c-myc shRNA vector was constructed successfully, in the normal presence of SDF-la, the level of TACRl-Tr mRNA in c-myc- cell group were lower than that in c-myc+ cell group( P < 0.05). But in the presence of SDF-la neutralized antibody, TACRl-Tr mRNA level of c-myc- cell group was higher than that of c-myc+ cell group(P < 0.05). Conclusion In the normal culture condition, c-MYC protein may transactivate TACRl-Tr transcription in MCF-7 cell, in the presence of SDF-1α neutral antibody, c-MYC protein lost the activity of transactivating for TACRl-Tr transciption.
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Objective To determine whether HBx gene can directly induce hepatocellular carcinoma in vivo, and to explore the mechanism of transplantation tumor in nude mice.Methods pCMVX/QSG7701 cell lines were vaccinated into subcutaneous tissue of nude mice. pRcCMV2/QSG7701 and QSG7701 cell lines were used as controls. The sections of transplantation tumor were observed microscopically by HE staining. RT-PCR was used to detect the expression of mutant p53 and c-Myc mRNA in transplant tumor and an other 3 cell lines. Results The transplant tumor occurred within the subcutaneous tissue of the nude mice inoculated with pCMVX/QSG7701 cell lines at 2nd week after the vaccination. No metastatic tumor was found in other organs. Transplant tumor was not formed in all the controls. HE staining confirmed that the transplant tumor was hepatocellular carcinoma. The mutant p53 mRNA and c-Myc mRNA expression level of transplant tumor and pCMVX/QSG7701 cells was significantly higher than that of pRcCMV2/QSG7701 and QSG7701 cells, respectively (P＜0.01).Conclusion HBx gene can up-regulate the expression of mutant p53 and c-Myc genes, and directly induce hepatocellular carcinoma in vivo.
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Objectives To evaluate the potential of antisense c myc gene therapy for acute promyelocytic leukemia by investigating the biological effects and molecular mechanisms in induction of differentiation of HL 60 cell line using recombinant antisense c myc adenovirus (Ad AS c myc). Methods Cultured HL 60 cells treated with Ad AS c myc or Ad LacZ with polybrene and protamine sulfate were analyzed by X gal staining, morphology, MTT, flow cytometric analysis, RT PCR, and immunocytochemical techniques in vitro . Results HL 60 cells could be transfected effectively by Ad LacZ+protamine sulfate (79.8%). The level of c myc transcription and the version could be strongly inhibited by Ad AS c myc in the transfected HL 60 cells. Ad AS c myc could strongly inhibit the cell growth in HL 60 cells (51%). Ad AS c myc could reduce the ratio of nuclear/cytoplasm, and increase the activity of peroxidase in HL 60 cells. Ad AS c myc could lead to the blocking of G 0/G 1 phase in HL 60 cells. Ad AS c myc could also increase the expression of c fos in HL 60 cells. Conclusion The expression of Ad AS c myc can inhibit the growth and induce differentiation of HL 60 cells in vitro . The biological effects of Ad AS c myc may be closely associated with the activity of peroxidase and c fos gene in HL 60 cells. Ad AS c myc is of clinical potential in gene thera py for acute promyelocytic leukemia.
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Objective:To evaluate the biological functions and underlying mechanisms of transcription factor PAX5 in promoting cell proliferation and anti-apoptosis in multiple myeloma cells.Methods:a PAX5 knockdown cell line was established by stable transfection of vector-based PAX5-siRNA into multiple myeloma IM9 cells.The expressing level of PAX5,p53,XBP-1 and c-Myc was examined by either or both Western blot and RT-PCR for the targets.Cell proliferation and apoptosis were assayed by MTT and flow cytometry,respectively.Results:PAX5 was expressed selectively in IM9 cells,but neither in another common used multiple myeloma KAS6 cells nor in the prostate cancer cells DU145 and PC3.Knockdown PAX5 led to a significant up-regulation of p53 and XBP1,but decreased c-Myc expressions in IM9 cells that were correlated with the increased sensitivity of drug-induced apoptosis and decreased proliferation rates when compared to the control cells.Conclusion:PAX5 is specifically expressed in IM9 cells,which promotes cell proliferation and anti drug-induced apoptosis.In addition to inhibiting the expression of p53 and XBP-1,PAX5 is found to induce expression of oncogene c-Myc in IM9 cells,and this finding indicates an undiscovered signal pathway that may contributes to the malignancy of Multiple Myeloma cells.
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Through the improvement on determination of rhTNF-? bioassay, we established the formal procedure . Methods; We choose L929 cell for rhTNF-? bioassay determination. To identify the influence of rhTNF-? on the growth of HL-60 cells, various concentration and effecting time were studied. Results: The results showed that the inhibition of rhTNF-? on HL-60 cells based on dose and time. Northern Dot Blot and DNA electrophoresis showed c-myc gene was surpressed, and DNA was damaged by rhTNF-?. Conclusion: Autometic calculation is responsible for bioassay of rhTNF-?. rhTNF-? could inhibit the proliferation of HL-60 cells and its mechanism is different from doxirubicin.
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Objective To investigate the relationship of c-myc protein expression in skin hemangioma of infant with different stage,cell proliferative activity and apoptos is.Methods Proliferating cell nudear antigen(PCNA) and c-myc were detected by immunohistoc hemical staining,and apoptosis was detected using in situ cell death detection( TUNEL method) in 58 skin hemangioma specimens of infant.Results Among the 58 hemangiomas,the positive rate of c-myc protein was significantly higher in involuting hemangiomas than that in proliferative hemangiomas(P