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1.
The tumor promoters 12-O-tetradecanoyl-phorbol-13-acetate and teleocidin markedly enhanced the transformation of C3H 10T1/2 mouse fibroblasts when these cells were transfected with the cloned human bladder cancer c-rasH oncogene. Transfection studies with the drug resistance marker gpt and time course studies indicate that this enhancement is not simply an effect on the process of DNA transfection. These findings, together with parallel studies with NIH 3T3 fibroblasts, also indicate that the competence of animal cells for DNA transfection is a function of the recipient cell line, the transfected marker, and the growth conditions. Our findings suggest that during multistage carcinogenesis tumor promoters may complement the function of activated cellular oncogenes.  相似文献   

2.
Mouse tumors induced by gamma radiation are a useful model system for oncogenesis. DNA from such tumors contains an activated K-ras oncogene that can transform NIH 3T3 cells. This report describes the cloning of a fragment of the mouse K-ras oncogene containing the first exon from both a transformant in rat-2 cells and the brain of the same mouse that developed the tumor. Hybrid constructs containing one of the two pieces were made and only the plasmid including the first exon from the transformant gave rise to foci in NIH 3T3 cells. There was only a single base difference (G----A) in the exonic sequence, which changed glycine to aspartic acid in the transformant. By use of a synthetic oligonucleotide the presence of the mutation was demonstrated in the original tumor, ruling out modifications during DNA-mediated gene transfer and indicating that the alteration was present in the thymic lymphoma but absent from other nonmalignant tissue. The results are compatible with gamma radiation being a source of point mutations.  相似文献   

3.
4.
The v-abl oncogene of the Abelson murine leukemia virus (A-MuLV) is known to efficiently transform NIH/3T3 fibroblasts in vitro and to cause an acute lymphosarcoma in susceptible murine hosts. The role of its relative, the bcr/abl gene product, in the etiology of human chronic myelogenous leukemia (CML) remains speculative. To assess the transforming properties of the bcr/abl gene product, complementary DNA clones encoding the CML-specific P210 bcr/abl protein were expressed in NIH/3T3 fibroblasts. In contrast to the v-abl oncogene product P160, the P210 bcr/abl gene product did not transform NIH/3T3 cells. Cell lines were isolated that expressed high levels of the P210 bcr/abl protein but were morphologically normal. During the course of these experiments, a transforming recombinant of bcr/abl was isolated which fuses gag determinants derived from helper virus to the NH2-terminus of the bcr/abl protein. This suggests that a property of viral gag sequences, probably myristylation-dependent membrane localization, must be provided to bcr/abl for it to transform fibroblasts.  相似文献   

5.
Blym-1, a transforming gene detected by transfection of NIH 3T3 cells with DNA from Burkitt lymphomas, was mapped to the short arm of chromosome 1 (1p32) by chromosomal in situ hybridization. The Blym-1 gene was not physically linked to the cellular myc oncogene or to any of the immunoglobulin gene loci implicated in the characteristic chromosomal translocations in Burkitt lymphoma.  相似文献   

6.
New method for detecting cellular transforming genes   总被引:24,自引:0,他引:24  
Tumor induction in athymic nude mice can be used to detect dominant transforming genes in cellular DNA. Mouse NIH 3T3 cells freshly transfected with either cloned Moloney sarcoma proviral DNA or cellular DNA's derived from virally transformed cells induced tumors when injected into athymic nu/nu mice. Tumors were also induced by cells transfected with DNA from two tumor-derived and one chemically transformed human cell lines. The mouse tumors induced by human cell line DNA's contained human DNA sequences, and DNA derived from these tumors was capable of inducing both tumors and foci on subsequent transfection. Tumor induction in nude mice represents a useful new method for the detection and selection of cells transformed by cellular oncogenes.  相似文献   

7.
erbB-2 is a potent oncogene when overexpressed in NIH/3T3 cells   总被引:94,自引:0,他引:94  
A wide variety of human tumors contain an amplified or overexpressed erbB-2 gene, which encodes a growth factor receptor-like protein. When erbB-2 complementary DNA was expressed in NIH/3T3 cells under the control of the SV40 promoter, the gene lacked transforming activity despite expression of detectable levels of the erbB-2 protein. A further five- to tenfold increase in its expression under influence of the long terminal repeat of Moloney murine leukemia virus was associated with activation of erbB-2 as a potent oncogene. The high levels of the erbB-2 product associated with malignant transformation of NIH/3T3 cells were observed in human mammary tumor cells that overexpressed this gene. These findings demonstrate a new mechanism for acquisition of oncogenic properties by genes encoding growth factor receptor-like proteins and provide a functional basis for the role of their overexpression in the development of human malignancies.  相似文献   

8.
C(3)H mice, made chimeric by lethal x-irradiation followed by injection of (C(3)H x T(6))F(1) spleen cells, were later stimulated by CCl(4) to produce a vigorous burst of hepatic parenchymal cell mitoses. Cytogenetic studies of the regenerating livers of 11 chimeras identified 89 percent of the cells as donor type by the presence of the distinctive T(6) marker.  相似文献   

9.
A monoclonal antibody was used to show directly positive thymic selection of the T cell repertoire in mouse strains expressing the 17a beta-chain variable domain (V beta 17a) of the T cell receptor. In the absence of the potent tolerizing class II major histocompatibility complex (MHC) molecule, I-E, peripheral expression of V beta 17a+ T cell receptors varied with the MHC haplotype of the mouse strain. In the most extreme case, H-2q mice expressed high peripheral levels of CD4+ V beta 17a+ T cells (14 to 19 percent), whereas H-2b mice expressed low levels (3 to 4 percent). Analysis of (b x q)F1 mice and chimeric mice showed that these differences were determined by positive thymic selection and implicated the thymic epithelium as the controlling cell type.  相似文献   

10.
In a study of the relation between chronic inflammation and carcinogenesis, C3H mouse fibroblasts of the 10T 1/2 clone 8 line (10T 1/2 cells) were exposed to human neutrophils stimulated to synthesize reactive oxygen intermediates or to a cell-free enzymatic system generating superoxide (xanthine oxidase plus hypoxanthine). After exposure, the 10T 1/2 cells were either placed in tissue culture or immediately injected into athymic nude mice. Both malignant and benign tumors developed in the mice injected with treated cells, but not in those injected with control cells; in one instance cells grown from one of the benign tumors subsequently developed a malignant phenotype. Malignant transformation was also observed in treated cells in the experiments in vitro.  相似文献   

11.
A model for human lymphocyte ontogeny has been developed in a normal mouse. Human bone marrow, depleted of mature T and B lymphocytes, and bone marrow from mice with severe combined immunodeficiency were transplanted into lethally irradiated BALB/c mice. Human B and T cells were first detected 2 to 4 months after transplantation and persisted for at least 6 months. Most human thymocytes (30 to 50 percent of total thymocytes) were CD3+CD4+CD8+. Human immunoglobulin was detected in some chimeras, and a human antibody response to dinitrophenol could be generated after primary and secondary immunization.  相似文献   

12.
alpha-Difluoromethyl ornithine and mouse type 1 interferon, when administered simultaneously, were highly toxic to B16 melanoma cells in culture. Oral administration of alpha-difluoromethyl ornithine suppressed B16 melanoma development in mice 85 percent whereas interferon given subcutaneously inhibited tumor growth only 24 percent. Total or near total suppression of tumor growth was observed in mice receiving both treatments.  相似文献   

13.
【目的】研究鹅卵泡基质层TLR家族基因表达谱及其对LPS不同处理时间后的反应性。【方法】在大群散养条件下,实时观察鹅产蛋过程,分别在产蛋后8和2 h以及产前4、16和28 h注射LPS,并在产蛋后8h屠宰,即LPS作用0、6、12、24和36 h,每个时间点各5只鹅。屠宰时,取卵巢,观察卵泡外观形态,并分离F1-F5级卵泡基质层。试验鹅自由饮水、自由采食、自然光照。LPS处理0、24和36 h的单个F1-F5级卵泡基质层或卵泡用于基因表达分析,而其他LPS处理的每只鹅等级卵泡基质层RNA等质量混合后用于基因表达分析。采用普通PCR方法检测10种鸟类TLR家族基因的在鹅等级卵泡基质层的表达谱,其中以脾脏组织为阳性对照,以不加c DNA模板为阴性对照。根据RT-PCR的表达谱结果,采用Real-time PCR检测不同等级卵泡基质层TLRs表达水平以及不同LPS处理时间对基质层TLRs表达水平的影响。对不同等级卵泡和不同LPS处理时间对基质层TLRs表达的数据,采用单因子方差分析,Duncan法多重比较;对变性卵泡与对照组基质层TLRs表达水平的数据,采用T检验分析。【结果】(1)已公开的10种鸟类TLR家族基因,如TLRs 1A、1B、2A、2B、3、4、5、7、15和21基因均在等级卵泡基质层组织中表达。(2)随等级卵泡生长,TLRs 2A和15表达水平逐渐升高;F1级卵泡基质层TLR2A表达水平显著高于F3、F4和F5级卵泡,TLR15表达水平显著高于F5级卵泡;其他TLR家族基因,如TLRs 1A、1B、2B、3、4、5、7和21在不同等级卵泡基质层中的表达水平差异不显著。(3)在LPS处理0、6和12 h时,等级卵泡外观形态无明显变化,但在LPS处理24 h时,3只鹅的等级卵泡呈深黄色胶冻状变性;在LPS处理36 h时,全部试验鹅等级卵泡呈深黄色胶冻状变性。(4)与对照组(LPS处理0 h)相比,TLRs 2A、4和5表达水平在LPS处理12和24 h时显著升高,TLR2B表达水平在LPS处理24 h时显著升高,TLRs 7和15表达水平在LPS处理6-24 h期间均显著升高,而TLRs 1A、1B、3和21表达水平在LPS处理6-24 h期间差异不显著。(5)与等级卵泡基质层相比,在LPS处理24和36 h的变性卵泡中,TLRs 1A、2A、2B、4、5、7和15表达水平显著升高,TLR3表达水平显著降低,而在LPS处理36 h的变性卵泡中,TLRs 1B和21表达水平显著升高。【结论】已发现的10种鸟类TLR家族基因均在种鹅等级卵泡基质层表达,且随LPS作用时间延长,等级卵泡形态结构发生变化,但TLRs表达水平逐渐升高。  相似文献   

14.
Production of urinary bladder carcinomas in mice by sodium saccharin   总被引:2,自引:0,他引:2  
Pellets weighing 20 to 24 milligrams and containing 20 percent sodium saccharin suspended in cholesterol were surgically implanted into the urinary bladder lumens of female Swiss mice (60 to 90 days old) under ether anesthesia. Incidences of mouse bladder carcinomas in animals exposed to these pellets were 47 and 52 percent as compared with incidences of 13 and 12 percent in control mice exposed to pellets of pure cholesterol. The exposure of the mouse bladder to saccharin was very brief, because the time required for 50 percent of the compound to be eluted from the pellets was about 5.5 hours.  相似文献   

15.
Rapid loss of tolerance induced in weanling NZB and B-W F1 mice   总被引:8,自引:0,他引:8  
Young adult mice (30 to 43 days old) of autoimmune NZB and BIW F(1) strains failed to develop immunologic tolerance when treated with ultracentrifuged bovine gamma globulin and then challenged 12 days later. By contrast, weanling NZB and B/ W F(1) mice (18 to 25 days) as well as weanling C3H mice (16 to 19 days) became tolerant and had no serum antibody 12 days after challenge. The C3H mice remained unresponsive, but NZB and B! W F(1) mice produced antibody between days 27 and 41. The rapid loss of previously established tolerance to foreign antigens could have its parallel in the loss of tolerance to autoantigens with subsequent development of lupus nephritis and Coombs' positive hemolytic anemia in these animals.  相似文献   

16.
Treatment of mice with the carcinogen N-methylnitrosourea results in the development of thymic lymphomas with frequent involvement of the N-ras oncogene. The activated mouse N-ras gene was isolated from one of these lymphomas and, by transformation in concert with restriction digestion, a map of the gene was prepared and its approximate boundaries were determined. By means of somatic cell hybrids the normal N-ras gene was found to be unlinked to other members of the ras gene family.  相似文献   

17.
【目的】明确F3代波杂山羊XKR4基因多态性与生长性状的关联性,筛选出优异基因为后期培育新品系提供理论依据。【方法】以F3代波杂山羊为研究对象,通过血液DNA混池测序鉴定XKR4基因SNP位点,统计SNP位点的等位基因频率、基因型频率、遗传纯合度(Ho)、期望杂合度(He)、有效等位基因数(Ne)及多态信息含量(PIC)等,以卡方(χ2)检验分析基因型是否处于Hardy-Weinberg平衡状态,并利用SPSS 25.0分析不同基因型与F3代波杂山羊生长性状的关联性。【结果】 F3代波杂山羊XKR4基因Intron-1区域存在2个SNPs位点,分别是C194069A和T194091A。C194069A位点的等位基因频率中C>A,优势基因型为CC型; T194091A位点的等位基因频率中T>A,优势基因型为TT型;C194069A和T194091A位点均处于Hardy-Weinberg平衡(P>0.05)。关联分析结果表明:在初生群体中,T194091A位点AA基因型与其他基因型的体斜长存在显著差异(P<0.05,下同);在3月龄群体中,C194069A和T194091A位点各基因型的生长性状指标间均无显著差异(P>0.05,下同);在6月龄群体中,C194069A和T194091A位点AA基因型的体重分别与相应位点的CC基因型和TT基因型存在显著差异;在12月龄群体中,C194069A位点AA基因型在体重和胸围方面均极显著优于CC基因型(P<0.01,下同),在体高方面与CC基因型存在显著差异,T194091A位点AA基因型的体重、体斜长分别与TT基因型呈显著或极显著差异。此外,在0~6月龄和0~12月龄,C194069A和T194091A位点AA基因型的日增重与CC基因型和TT基因型分别存在呈显著或极显著差异。【结论】 F3代波杂山羊XKR4基因存在2个SNPs位点(C194069A和T194091A),且均表现为AA基因型个体的生长性能优于其他基因型个体,即XKR4基因可作为F3代波杂山羊生长性状的候选基因。  相似文献   

18.
Cells of the mouse cell line 3T3-F442A can be induced by various hormones to differentiate into adipocytes, whereas cells of 3T3-C2, a subclone of 3T3, cannot. However, transfection of DNA from uninduced 3T3-F422A cells into 3T3-C2 cells permits recovery of 3T3-C2 transfectants that differentiate into adipocytes in the presence of insulin. DNA isolated from human fat tissue, when transfected into 3T3-C2 mouse cells, also gives rise to mouse transfectants that are induced to differentiate into adipocytes by the addition of insulin. Apparently, transfection of a trans-regulatory gene (or genes) from 3T3-F442A or human fat cells into 3T3-C2 cells is sufficient to commit 3T3-C2 cells to adipocyte differentiation.  相似文献   

19.
In this study, we introduced the bovine immunoglobulin μ heavy-chain gene (the orphaned gene on BTA11) into mouse germline cells. Bovine IgM was highly expressed in selected transgenic lines, and it largely inhibited rearrangements of the endogenous immunoglobulin heavy chain (IgH) genes in these lines. The forced expression of bovine IgM resulted in reduced numbers of pro- and pre-B cells but increased the number of immature B cells in the transgenic mice. Bovine IgM-expressing B cells can migrate from the bone marrow to the spleen, but most of the cells are arrested at the T1 transitional B cell stage, leading to a significantly lower number of T2 transitional and mature B cells in the spleen. Although the serum concentrations of endogenous IgM and IgG in the transgenic mice were significantly decreased, the IgA levels were slightly increased compared to the WT mice. The bovine IgM level in the serum was only one-tenth to one-fifth of that of endogenous mouse IgM, suggesting that most of the serum immunoglobulin were contributed by endogenous IgH gene-expressing B cells. These transgenic mice also exhibited a lower frequency of unique complementarity determining region 3 (CDR3) sequences in their VH repertoire and Vκ repertoire but exhibited an increased frequency of unique CDR3 in their Vλ repertoire. Compared to the WT mice, the transgenic mice had a significantly higher percentage of mouse IgM-expressing B cells that expressed λ chains. Finally, we showed that the transgenic mice were deficient in a specific antibody response to antigen stimulation.  相似文献   

20.
Neurotransmitter receptors are usually restricted to neuronal cells, but the signaling pathways activated by these receptors are widely distributed in both neural and non-neural cells. The functional consequences of activating a brain-specific neurotransmitter receptor, the serotonin 5HT1c receptor, in the unnatural environment of a fibroblast were examined. Introduction of functional 5HT1c receptors into NIH 3T3 cells results, at high frequency, in the generation of transformed foci. Moreover, the generation and maintenance of transformed foci requires continued activation of the serotonin receptor. In addition, the injection of cells derived from transformed foci into nude mice results in the generation of tumors. The serotonin 5HT1c receptor therefore functions as a protooncogene when expressed in NIH 3T3 fibroblasts.  相似文献   

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