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Intratumoral expression of genes encoding Cytochrome P450 enzymes (CYP) might play a critical role not only in cancer development but also in the metabolism of anticancer drugs. The purpose of this study was to compare the mRNA expression patterns of seven representative CYPs in paired tumor and normal tissue of child patients with rabdomyosarcoma (RMS). Using real time quantitative RT-PCR, the gene expression pattern of CYP1A1, CYP1A2, CYP1B1, CYP2E1, CYP2W1, CYP3A4, and CYP3A5 were analyzed in tumor and adjacent non-tumor tissues from 13 child RMS patients. Protein concentration of CYPs was determined using Western blot. The expression levels were tested for correlation with the clinical and pathological data of the patients. Our data showed that the expression levels of CYP1A1 and CYP1A2 were negligible. Elevated expression of CYP1B1 mRNA and protein was detected in most RMS tumors and adjacent normal tissues. Most cancerous samples exhibit higher levels of both CYP3A4 and CYP3A5 compared with normal tissue samples. Expression of CYP2E1 mRNA was found to be significantly higher in tumor tissue, however no relation was found with protein levels. CYP2W1 mRNA and/or protein are mainly expressed in tumors. In conclusion, we defined the CYP gene expression profile in tumor and paired normal tissue of child patients with RMS. The overexpression of CYP2W1, CYP3A4 and CYP3A5 in tumor tissues suggests that they may be involved in RMS chemoresistance; furthermore, they may be exploited for the localized activation of anticancer prodrugs.  相似文献   

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Resveratrol (RSV) has a beneficial role in the prevention of diabetes and alleviates some diabetic complications, such as cardiomyopathy. We investigated cyclooxygenase-1 (COX-1), COX-2, nuclear factor κB (NF-κB), matrix metalloproteinase-9 (MMP-9), and sirtuin 1 (SIRT1) mRNA expression levels in heart tissue after RSV treatment in streptozotocin (STZ)-induced diabetic rats. After induction of chronic diabetes with STZ, 10 mg RSV/kg per day was administered to DM and DM+RSV groups for four weeks. At the end of the experiment, all rats were sacrificed and heart tissues were stored at -80°C; mRNA expression levels of COX-1, COX-2, NF-κB, MMP-9, and SIRT1 genes were analyzed with quantitative real-time PCR. We did not find any significant effect of RSV on MMP-9, COX-1, COX-2, or NF-κB mRNA levels among the groups. However, SIRT1 mRNA levels decreased in the DM group compared to controls and increased in the DM+RSV group when compared to the DM group. SIRT1 is activated by RSV treatment in diabetic heart tissue. Activation of SIRT1 by RSV may lead to a new therapeutic approach for diabetic heart tissue. We conclude that RSV treatment can alleviate heart dysfunction by inhibiton of inflammatory gene expression such as SIRT1.  相似文献   

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人类抑癌基因beclin 1在胃癌和直结肠癌中表达下调的研究   总被引:1,自引:0,他引:1  
人类抑癌基因beclin 1通过自噬作用调节细胞生长,但在胃癌和直结肠癌中其表达水平和调控机制仍不清楚.通过检测胃癌和直结肠肿瘤组织中beclin 1基因的表达水平,及DNA异常甲基化和杂合子缺失对其表达的影响,发现与癌旁组织相比,35%的胃癌标本和30%的直结肠癌标本中beclin 1基因表达显著下调.同时发现,beclin 1基因5’端存在一高密度CpG岛,在胃癌和直结肠癌中beclin 1的启动子区域和第二个内含子区域存在甲基化,而杂合子缺失仅在胃癌中发生.这些发现表明beclin 1基因的异常甲基化和杂合子缺失对其在胃癌和直结肠癌中的表达起调控作用.  相似文献   

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JunB is essential for mammalian placentation   总被引:15,自引:0,他引:15       下载免费PDF全文
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We have investigated the effect of E26, an avian leukemia retrovirus, on the growth properties of chicken embryo fibroblasts (CEFs). E26-infected CEFs were not transformed, according to several transformation parameters, but exhibited an activated growth in vitro. They started to grow without latency in serum-supplemented medium, maintained long-term growth in regular or low-serum medium, and could grow when seeded at low cell density in low-serum medium. We compared the integration and the level of expression of the proviral DNA in E26-infected CEFs and E26-transformed hematopoietic cells. An average of two provirus copies were found in each kind of cells. However, whereas high contents of both viral mRNA and E26-specific protein products were found in transformed hematopoietic cells, we detected only low amounts of viral mRNA and no E26 protein in infected CEFs. These data show that the level of expression of the E26 provirus is lower in CEFs than in hematopoietic cells. They suggest that transformation efficiency of the virus depends on its level of expression.  相似文献   

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The wnt signaling pathway has important functions in nervous system development. To better understand this process we have cloned and analyzed the expression of the wnt receptor, frizzled 9, in the developing nervous system in mouse, chick and zebrafish. The earliest expression of mouse frizzled 9 mRNA expression begins at E8.5 with expression throughout the entire rostral-caudal neuraxis. This early expression pattern within the neural tube appears to be conserved between chick and zebrafish. Expression becomes restricted to a ventral domain in the mouse ventricular zone at E11.5, a region specified to give rise to neurons and glia. Using a polyclonal antibody to MFZ9 further shows expression limited to neural restricted precursors cells.  相似文献   

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We have used specific cloned cDNA probes generated from the mRNA coding for the vitamin D-induced 28,000-Da chick intestinal calcium binding protein (calbindin) to study the hormonal regulation of the expression of this mRNA by 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3]. The calbindin-mRNA has been analyzed in chicken intestinal poly(A)+ mRNA samples as well as other chicken tissues by "Northern" blot analysis. There exists a predominant mRNA species of approximately 2000 nucleotides and two minor cross-hybridizing species that are nearly equivalent in proportion; their sizes are approximately 2600 and 3100 nucleotides. All three mRNA species are nonexistent in the chick intestine in the absence of vitamin D3 intake. However, all three mRNA species begin to accumulate at the same time in the chick intestine following the administration of the hormonally active metabolite of vitamin D3, 1,25-(OH)2D3. This response in the intestine is very similar to other steroid hormone-regulated gene products. All three mRNA species exist in the cell cytoplasm and are present on soluble polysome complexes, suggesting that all three are engaged in protein synthesis. Examination of other chick tissues (both vitamin D-deficient and -replete) reveals a close association between mRNA expression and previously observed calbindin expression. Each tissue is unique in the steady-state level of expression of the calbindin-mRNAs.  相似文献   

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Expression of the gene coding for a new breast tumor-associated antigen, H23, was compared to expression of genes coding for pS2, c-erbB2 and estrogen receptor (ER). Comparison involved mRNA expression in normal and malignant breast tissues as well as in non-breast tumors. Results obtained by RNA dot blot and Northern hybridizations showed that expression of the H23 antigen coding gene is a discriminatory marker in human breast cancer. It is expressed in 92% of breast tumors whereas 69%, 62% and 56% of breast tumors demonstrate significant mRNA levels of c-erbB2, ER and pS2, respectively. Non-malignant or normal breast tissue expresses much lower levels of the H23 antigen mRNA. From the comparative analysis presented here it is concluded that the gene coding for H23 antigen furnishes a most useful marker for human breast cancer.  相似文献   

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We previously demonstrated that Avian Leukemia Viruses (ALV) carrying the v-myc gene specifically induce two types of tumors, cardiomyocytic tumors when the virus is injected before embryonic day 3 (E3), skin tumors when the virus is injected at E3 or E5.

Aiming to elucidate the mechanisms which determine this time-dependent change in target, we infected chick and quail embryos at E3 and E5 with replication-deficient, lacZ gene-carrying, ALV-based viruses produced by a packaging cell line. Three constructs driven by 3 different Long Terminal Repeats (LTRs) were tested and yielded similar results. When the constructs were inoculated at E3 and the lacZ gene product revealed 5 days later, around 70% of the embryos carried lacZ+ clones in the heart, around 50% had positive clones in the skin anywhere on the body, while a few embryos displayed clones in internal organs (liver, stomach, lungs). Immunocytological identification of the heart cell type(s) expressing the virus revealed that the only cells infected were cardiomyocytes. When the constructs were inoculated at E5, no lacZ+ clones appeared in the heart but all were located in the cephalic skin. In order to examine the relationship between viral integration and expression, DNA of different organs or tissues from lacZ stained embryos was analyzed by PCR. A tight correlation between integration and expression in the heart and in the skin was revealed in most cases. In contrast, a significant PCR signal was often detected in the liver or the stomach despite weak or absent expression as revealed by lacZ+ clones.

We then investigated the influence of envelope glycoprotein subgroups on the tropism of these constructs. The lacZ vector driven by RAV-2 LTRs was packaged as subgroups A, B or E viral particles. The A subgroup, used in the part of the study described above, infects both chick and quail while the B and E subgroups are specific for chick or quail respectively. These B and E subgroups induced lacZ+ clones in the heart (after E3 injection) while no clones or only a few were detected in the skin either after E3 or E5 injection. The following conclusions can be drawn: 1) cardiomyocytes are at E3 the major target for integration and expression of ALV-derived viruses in vivo; 2) targets change rapidly with embryonic age; and 3) tissue-specific infections depend on the envelope subgroup, thus presumably on the presence of the cognate receptor. This study clearly indicates that E3 inoculation of ALV-based retroviral vectors is a simple and powerful method to transfer gene sequences into cardiomyocytes and epidermal cells.  相似文献   

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HMP19 is a neuron-specific gene; its expression product belongs to a family of neuronal proteins which can be found in numerous kinds of human cancers. However, the clinicopathological significance of HMP19 expression in epithelial ovarian cancer (EOC) is as yet unknown. In this study, protein expression levels of HMP19 in cancerous tissues were determined by tissue microarray immunohistochemistry analysis (TMA-IHC) (n = 117). HMP19 protein levels in cancer tissues were associated with clinical characteristics and overall survival rates of patients with EOC. It was found that both mRNA and protein levels of HMP19 were significantly lower in EOC than those in normal ovary or fallopian tube tissues (P<0.05). The protein expression level of HMP19 was significantly associated with a lower FIGO stage, a lower level of CA-125 and a lower presence of metastasis. Consistent with related adverse clinical pathological features, the overall survival (OS) rate of patients with low or non HMP19-expressing tumors was inferior compared to those with high HMP19-expressing tumors. This is in accordance with further studies that found high HMP19 protein level to be an independent prognostic factor for OS in EOC. Multivariate analysis demonstrated that tumor patients with low HMP19 expression had an exceedingly poor OS. HMP19 plays a role in metastasis/tumor suppression and offers a prognostic value for EOC. HMP19, as a new inhibitor, strongly inhibits metastasis and partially attenuates tumor growth in EOC.  相似文献   

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Proteins binding to the PEA3 enhancer motif (AGGAAG) activate the polyomavirus early promoter and help comprise the viral late mRNA initiator element (W. Yoo, M. E. Martin, and W. R. Folk, J. Virol. 65:5391-5400, 1991). Because many developmentally regulated cellular genes have PEA3 motifs near their promoter sequences, and because Ets family gene products activate the PEA3 motif, we have studied the expression of PEA3-binding proteins and Ets-related proteins during differentiation of F9 embryonal carcinoma cells. An approximately 91-kDa protein (PEA3-91) was identified in F9 cell nuclear extracts by UV cross-linking to a radiolabeled PEA3 oligonucleotide probe, and expression of PEA3-91 was down-regulated after differentiation of F9 cells to parietal endoderm. The c-ets-1 gene product binds to a sequence in the murine sarcoma virus long terminal repeat that is similar to the PEA3 motif (cGGAAG), but PEA3-91 was not cross-linked to this Ets-1-binding motif, nor did antiserum which recognizes murine c-ets-1 and c-ets-2 proteins have any effect on PEA3-binding activity in mobility shift assays. Furthermore, c-ets-1 mRNA was not detected in undifferentiated or differentiated F9 cells, and c-ets-2 mRNA levels remained high after differentiation. Antiserum against the Drosophila Ets-related E74A protein, however, recognized an approximately 92-kDa protein in F9 cells whose expression during differentiation varied in a manner identical to that of PEA3-91. These data suggest that PEA3-91 is not the product of the ets-1 or ets-2 genes but is likely to be the product of a murine homolog of the Drosophila E74 gene.  相似文献   

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