RESUMO
<p><b>OBJECTIVE</b>To study the expression of MUM-1/IRF4 and its significance in follicular lymphoma.</p><p><b>METHODS</b>Ninety-eight cases of follicular lymphoma were enrolled into the study. They were graded according to the 2008 WHO criteria. The expression of MUM-1/IRF4 protein and other markers (CD10, bcl-6, bcl-2 and Ki-67) was studied using tissue microarray and immunohistochemistry.</p><p><b>RESULTS</b>Amongst the 98 cases studied, there were 24 grade 1 cases, 30 grade 2 cases, 26 grade 3A cases and 18 were grade 3B cases. The rates of expression of MUM-1/IRF4, CD10, bcl-6, bcl-2 and Ki-67 (≥ 25%) were 39.8% (39/98), 62.2% (61/98), 80.6% (79/98), 87.8% (86/98) and 50.0% (49/98), respectively. MUM-1/IRF4 predominantly expressed in high-grade follicular lymphoma and showed a significantly positive correlation with lymphoma grade (r = 0.628, P = 0.000) and Ki-67 index (r = 0.473, P = 0.000). MUM-1/IRF4 expression had a significantly negative correlation with CD10 expression (r = -0.597, P = 0.000), but no correlation with bcl-6 and bcl-2 expression.</p><p><b>CONCLUSIONS</b>MUM-1/IRF4 expression is significantly higher in high-grade follicular lymphoma, indicating that these cases have a high proliferative activity, more aggressive behavior and poorer prognosis. MUM-1/IRF4, when strongly expressed, is another helpful marker for the diagnosis of high-grade follicular lymphoma.</p>
Assuntos
Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Biomarcadores Tumorais , Metabolismo , Proteínas de Ligação a DNA , Metabolismo , Fatores Reguladores de Interferon , Metabolismo , Antígeno Ki-67 , Metabolismo , Linfoma Folicular , Metabolismo , Patologia , Gradação de Tumores , Neprilisina , Metabolismo , Proteínas Proto-Oncogênicas c-bcl-2 , Metabolismo , Proteínas Proto-Oncogênicas c-bcl-6RESUMO
<p><b>OBJECTIVE</b>To investigate the sensitizing potential of Shuanghuanglian Injection (SHL) by comparing the popliteal lymph node (PLN) response in mice induced by SHL and chemicals.</p><p><b>METHODS</b>Sixty female C57BL/6J mice were equally and randomly divided into six groups, i.e. the blank control group (A) and five treated groups treated respectively with phenobarbital 1 mg/mouse (B), mercuric chloride ( HgCl2) 50 microg/mouse (C), D-penicillamine 2 mg/mouse (D), and SHL in low (1 mg/mouse) and high (5 mg/mouse) dosages (E and F) via subcutaneous injection into left pad of hind foot. Animals were sacrificed on the 8th day after injection, their bilateral PLNs were isolated and weighed respectively to calculate the PLN mass index (MI). Then the PLNs get from four mice in each group were fixed with 4% paraformaldehyde solution for histopathologic examination; the other six PLNs were prepared into single-cell suspensions to calculate cell index (CI) for comparing the changes of PLN in various groups.</p><p><b>RESULTS</b>MI and CI in Group F reached to > or = 2 and > or = 5 (average) respectively, which was higher than those in Group A (P<0.05). Pathological examination showed that the left PLN in Group F enlarged, with remarkable germinal center and increased high endothelial venules proliferation.</p><p><b>CONCLUSION</b>SHL could induce significant PLN response in C57BL/6J mice, suggesting it has certain sensitizing potential.</p>
Assuntos
Animais , Feminino , Camundongos , Medicamentos de Ervas Chinesas , Hipersensibilidade , Patologia , Ensaio Local de Linfonodo , Linfonodos , Alergia e Imunologia , Patologia , Camundongos Endogâmicos C57BLRESUMO
<p><b>OBJECTIVE</b>To reverse the multidrug resistant (MDR) phenotype of human breast carcinoma cells by small hairpin RNA (shRNA) technique targeting hypoxia-inducible factor (HIF)-1alpha gene.</p><p><b>METHODS</b>Small hairpin RNA (shRNA) eukaryotic expression vector targeting HIF-1alpha gene, named pSilencer-HIF, was constructed and transfected into MCF-7/ADR human breast cancer cells by liposome technique. Tumor cell livability (TCL) and Rhodamine 123 efflux assay were used to monitor the biological changes of the transfected cells. The mRNA and protein expression of HIF-1alpha and mdr-1 were investigated by RT-PCR and Western blot.</p><p><b>RESULTS</b>The successful construction of pSilencer-HIF plasmid was confirmed by DNA sequencing. HIF-1alpha mRNA and protein levels were significantly decreased in MCF-7/ADR cells after the transfection and there was a direct correlation between HIF-1alpha and mdr-1 expression. By comparing the cells transfected with control vector and the MCF-7/ADR cells transfected with pSilencer-HIF, a reduced TCL from 76% to 43%, and an increased Rhodamine 123 fluorescence intensity from 22.0% to 86.6% were observed.</p><p><b>CONCLUSIONS</b>pSilencer-HIF-1alpha has been successfully constructed. The inhibition of HIF-1alpha expression through shRNA technique can significantly reverse the multidrug resistance phenotype of MCF-7/ADR cells.</p>
Assuntos
Humanos , Neoplasias da Mama , Patologia , Linhagem Celular Tumoral , Resistência a Múltiplos Medicamentos , Fisiologia , Resistencia a Medicamentos Antineoplásicos , Subunidade alfa do Fator 1 Induzível por Hipóxia , Interferência de RNA , RNA Interferente Pequeno , FarmacologiaRESUMO
<p><b>OBJECTIVE</b>To study the relationship of expression of mucin 1 and E-cadherin with recurrence of pleomorphic adenoma in salivary gland, and to investigate the signal to predict the recurrence potential of the tumor.</p><p><b>METHODS</b>; The capsule of tumor was observed by microscope. The expression of mucin 1 and E-cadherin in 33 cases of primary adenoma, 12 cases of recurrent pleomorphic adenomas and 7 cases of malignant pleomorphic adenomas were detected by immunohistochemistry.</p><p><b>RESULTS</b>There was no significant difference about the status of capsule and the positive rate of mucin 1 expression between primary and recurrent pleomorphic adenoma (P > 0.05). The abnormal distribution of mucin 1 expression was observed in recurrent pleomorphic adenoma (6/8), which was characterized by the positive staining of the whole cytomembrane. On the other hand, positive staining of the primary pleomorphic adenoma was observed on the top of the membrane (19/21). The difference was statistically significant (P < 0.05). The staining pattern in malignant pleomorphic adenoma was similar with the recurrent ones except higher ratio of positive expression. No significant different was observed among the three kind of tumors on the expression rate of E-cadherin (P > 0.05).</p><p><b>CONCLUSION</b>The status of capsule didn't have much actual usage in predicting the recurrence of pleomorphic adenoma. There was no significant relationship between the expression of E-cd and the recurrence of the tumor. The abnormal distribution of mucin 1 expression contributes to the invasiveness of the tumor and can be used as the predictive signal for recurrence of pleomorphic adenoma.</p>
Assuntos
Humanos , Adenocarcinoma , Adenoma Pleomorfo , Caderinas , Metabolismo , Imuno-Histoquímica , Mucina-1 , Metabolismo , Recidiva Local de Neoplasia , Neoplasias das Glândulas SalivaresRESUMO
<p><b>OBJECTIVE</b>To stably reverse the multidrug resistance (MDR) of breast carcinoma cells in vitro.</p><p><b>METHODS</b>Two anti-mdr-1 ribozyme plasmids, RZ196 and RZ179, were constructed with EGFP as reporter gene and transfected into drug-resistant breast carcinoma cells in vitro. The expression of EGFP was observed by laser confocal microscopy. Flow cytometry, RT-PCR and Rhodamine123 efflux assay were used to detect P-glyco protein (p-gp) and mdr-1 mRNA.</p><p><b>RESULTS</b>After transfection with RZ196 and RZ179, the mdr-1 indices were reduced from 2.20 to 0.76 and 1.40, the expression rates of p-gp were reduced from 55.0% to 4.6% and 18.2%, the fluorescence intensity increased from 22.0% to 46.2% and 70.1%, TCL reduced from 75% to 28% and 43% respectively. In addition, the expression of ribozyme plasmid in tumor cells was stable under G418 selection. After two months, the mdr-1 indices remained at 0.81 and 1.47 in the cells transfected RZ196 and RZ179 respectively. The expression rates of p-gp were 5.2% and 19.5% and the Rh123 fluorescence intensity was 51.4% and 71.6% respectively.</p><p><b>CONCLUSIONS</b>Both anti-mdr-1 ribozyme RZ196 and RZ179 can stably reverse MDR phenotype of breast carcinoma cells in vitro. RZ196 construct appears to be more effective.</p>
Assuntos
Humanos , Membro 1 da Subfamília B de Cassetes de Ligação de ATP , Genética , Antineoplásicos , Farmacologia , Neoplasias da Mama , Genética , Patologia , Terapêutica , Doxorrubicina , Farmacologia , Resistência a Múltiplos Medicamentos , Resistencia a Medicamentos Antineoplásicos , Técnicas de Transferência de Genes , Genes MDR , Genética , Vetores Genéticos , RNA Catalítico , Genética , Retroviridae , GenéticaRESUMO
<p><b>OBJECTIVE</b>To reverse the multidrug resistance (MDR) property of carcinoma cells by blocking transcription of activating sites of mdr-1.</p><p><b>METHODS</b>Breast carcinoma cells were transinfected with several antisense oligonucleotide (ASODN) complementary to mdr-1 by lipofectin. RT-PCR was used to detect the production of mdr-1mRNA. The expression of P-glycoprotein (gp) was then detected by immunohistochemistry and the function of P-gp was detected by rhodamine123 retention.</p><p><b>RESULTS</b>Forty-eight hours after transfection, mdr-1 index of cells treated by ASODN complementary to MA zone (major initiation start zone), MI (minor initiation start zone), C zone (CAAT box), G zone (GC box) of mdr-1 gene was 1.4, 1.9, 1.6 and 2.1 respectively. The rate of P-gp protein expression in treated cells was 14%, 43%, 26% and 39% respectively. The intracellular Rh123 retention in treated cells was 125%, 83%, 102% and 77% respectively. There was significant difference between cells treated by ASODN complementary to MA zone and C zone and drug-resistant cells.</p><p><b>CONCLUSIONS</b>The ASODN complementary to MA zone and C zone of mdr-1 gene can reverse MDR of drug-resistant cells to various extent, amongst which the former is more effective. Down-regulating transcription of mdr-1 by blocking transcription activating sites can reduce the expression of mdr-1mRNA and P-gp, and thus reversing MDR of carcinoma cells. The ASODN complementary to MI zone, G zone of mdr-1 however do not significantly reverse the MDR property.</p>
Assuntos
Humanos , Membro 1 da Subfamília B de Cassetes de Ligação de ATP , Genética , Neoplasias da Mama , Genética , Metabolismo , Patologia , Linhagem Celular Tumoral , Regulação para Baixo , Resistência a Múltiplos Medicamentos , Genética , Resistencia a Medicamentos Antineoplásicos , Genética , Regulação Neoplásica da Expressão Gênica , Imuno-Histoquímica , Oligonucleotídeos Antissenso , Genética , Metabolismo , RNA Mensageiro , Genética , Metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transcrição Gênica , GenéticaRESUMO
<p><b>OBJECTIVE</b>To study the reversing effect of Grape seed polyphenol (GSP) on multidrug resistance of MCF-7/ADR cell in vivo.</p><p><b>METHODS</b>The transplantable breast carcinoma cell line MCF-7/ADR model was established in BALB/C-nu/nu mice by subcutaneous implantation. Flow cytometry (FCM) was used to investigate the changes of Pgp expression and apoptosis rate after different drug treatment.</p><p><b>RESULTS</b>GSP has some effect on inhibition of tumor growth (the rate of inhibition was 18.35%), and combined with adriamycin can significantly inhibit tumor growth in nude mice, 20 mg/kg GSP can effectively reverse the resistance of MCF-7/ADR cells to ADR in vivo, and the rate of inhibition was 54.64%. FCM results showed that the expression of Pgp was significantly decreased (32.03 +/- 2.09) After administration of GSP and ADR, there was distinct difference between it and control (55.13 +/- 2.12). The mean rate of apoptosis was 15.12% +/- 1.04%, and was significantly increased compared with control (9.07% +/- 0.43%), P < 0.05.</p><p><b>CONCLUSION</b>GSP can effectively reversed the resistance of MCF-7/ADR cells in nude mice, the mechanism may be correlated with inhibition of Pgp expression and apoptosis.</p>
Assuntos
Animais , Feminino , Humanos , Camundongos , Membro 1 da Subfamília B de Cassetes de Ligação de ATP , Metabolismo , Antineoplásicos Fitogênicos , Farmacologia , Apoptose , Neoplasias da Mama , Metabolismo , Patologia , Linhagem Celular Tumoral , Doxorrubicina , Farmacologia , Interações Medicamentosas , Resistência a Múltiplos Medicamentos , Resistencia a Medicamentos Antineoplásicos , Flavonoides , Farmacologia , Camundongos Nus , Inoculação de Neoplasia , Fenóis , Farmacologia , Polifenóis , Sementes , VitisRESUMO
<p><b>OBJECTIVE</b>To assess the role of methylated mismatch repair (MMR) genes (hMLH1, hMSH2 and hMSH3) in the carcinogenesis and progression of hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>Samples of 38 cases of HCC along with their corresponding noncancerous tissues, 2 samples of donated normal tissue and 6 cell lines were collected and subject to the methylation-specific PCR (MSP) to examine promoter methylation status of MLH1, MSH2 and MSH3. Six tumor cell lines were analyzed before and after 5-aza-2'-deoxycytidine treatment. In addition, alterations of mRNA expression of MMRs were investigated by quantitative reverse transcription-PCR.</p><p><b>RESULTS</b>CpG island methylation of hMLH1 and hMSH2 was observed in 13.2% (5 of 38 samples) and 68.4% (26 of 38 samples) respectively in HCC, 2.6% (1 of 38 samples) and 55.3% (21 of 38) respectively in corresponding noncancerous tissues, but not in normal control tissues. Promoter methylation of the hMSH2 gene was present in 83.3% of cell lines tested (5/6), but none were observed for the hMLH1 gene. Promoter methylation of the hMSH3 gene was not identified in any tissue samples or cell lines. After 5-aza-2'-deoxycytidine treatment, hMSH2 methylation was induced or completely reversed, and its mRNA expression was increased in most cell lines.</p><p><b>CONCLUSIONS</b>Our results suggest that promoter hypermethylation of hMLH1 and hMSH2 genes is common in HCC. Particularly, there is a high frequency of methylation of hMSH2 in both cancer and noncancerous tissues, but not in normal control tissue. Therefore, hypermethylation of MMR genes, especially hMSH2, may be involved in the carcinogenesis of HCC and may serve as an early diagnostic marker for HCC. The close correlation between hMSH2 methylation and low expression of its mRNA suggests that hMSH2 methylation is an important pathway in the regulation of gene expression.</p>