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Adenovirus mediated p53 tumour suppressor gene therapy for human gastric cancer cells in vitro and in vivo 总被引:22,自引:0,他引:22 下载免费PDF全文
Ohashi M Kanai F Ueno H Tanaka T Tateishi K Kawakami T Koike Y Ikenoue T Shiratori Y Hamada H Omata M 《Gut》1999,44(3):366-371
BACKGROUND/AIMS: Gastric cancer is one of the most prevalent forms of cancer in East Asia. Point mutation of the p53 gene has been reported in more than 60% of cases of gastric cancer and can lead to genetic instability and uncontrolled cell proliferation. The purpose of this investigation was to evaluate the potential of p53 gene therapy for gastric cancer. METHODS: The responses of human gastric cancer cell lines, MKN1, MKN7, MKN28, MKN45, and TMK-1, to recombinant adenoviruses encoding wild type p53 (AdCAp53) were analysed in vitro. The efficacy of the AdCAp53 treatment for MKN1 and MKN45 subcutaneous tumours in nude mice was assessed in vivo. RESULTS: p53-specific growth inhibition was observed in vitro in two of four gastric cancer cell lines with mutated p53, but not in the wild type p53 cell line. The mechanism of the killing of gastric cancer cells by AdCAp53 was found, by flow cytometric analysis and detection of DNA fragmentation, to be apoptosis. In vivo studies showed that the growth of subcutaneous tumours of p53 mutant MKN1 cells was significantly inhibited by direct injection of AdCAp53, but no significant growth inhibition was detected in the growth of p53 wild type MKN45 tumours. CONCLUSIONS: Adenovirus mediated reintroduction of wild type p53 is a potential clinical utility in gene therapy for gastric cancers. 相似文献
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Synergistic interaction between hypergastrinemia and Helicobacter infection in a mouse model of gastric cancer 总被引:33,自引:0,他引:33
Wang TC Dangler CA Chen D Goldenring JR Koh T Raychowdhury R Coffey RJ Ito S Varro A Dockray GJ Fox JG 《Gastroenterology》2000,118(1):36-47
BACKGROUND & AIMS: Hypergastrinemia occurs frequently in association with acid suppression and Helicobacter infection, but its role in the progression to gastric atrophy and gastric cancer has not been well defined. METHODS: The effects of hypergastrinemia, and possible synergy with Helicobacter felis infection, were investigated in insulin-gastrin (INS-GAS) transgenic mice. RESULTS: INS-GAS mice initially showed mild hypergastrinemia, increased maximal gastric acid secretion, and increased parietal cell number but later progressed to decreased parietal cell number and hypochlorhydria. Development of gastric atrophy was associated with increased expression of growth factors, heparin-binding epidermal growth factor and transforming growth factor alpha. At 20 months of age, INS-GAS mice showed no evidence of increased enterochromaffin-like cell number, but instead exhibited gastric metaplasia, dysplasia, carcinoma in situ, and gastric cancer with vascular invasion. Invasive gastric carcinoma was observed in 6 of 8 INS-GAS mice that were >20 months old. Helicobacter felis infection of INS-GAS mice led to accelerated (< or = 8 mo) development of intramucosal carcinoma (85%), with submucosal invasion (54%) and intravascular invasion (46%; P < or = 0.05). CONCLUSIONS: These findings support the unexpected conclusion that chronic hypergastrinemia in mice can synergize with Helicobacter infection and contribute to eventual parietal cell loss and progression to gastric cancer. 相似文献
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Naoki Mori Makio Wada Jun Yokota Masaaki Terada Michiko Okada† Masanao Teramura Michihiko Masuda Shigeru Hoshino Toshiko Motoji Kazuo Oshimi Hideaki Mizoguchi 《British journal of haematology》1992,81(2):235-240
Mutations of the p53 tumour suppressor gene have frequently been observed in several types of solid tumours and are believed to be implicated in the development of these tumours. To determine the relevance of p53 mutations in haematologic neoplasms, we performed polymerase chain reaction-single strand conformation polymorphism analysis on the p53 gene in 45 patients with various types of haematologic neoplasms. In exons 5-8 containing highly conserved regions, mobility shifts indicating sequence alterations were detected in four of the 45 patients, and subsequent sequencing was performed. A point mutation resulting in a novel stop codon was detected at codon 213 in one of 23 cases of chronic myelogenous leukaemia (one of five cases of blast crisis). Point mutations causing amino acid substitutions were detected in one of four cases of myelodysplastic syndrome at codon 195, one of three cases of adult T-cell leukaemia at codon 281, and one of eight cases of acute lymphoblastic leukaemia at codon 281, and these missense mutations were accompanied by loss of the wild type allele. Patients harbouring these nonsense and missense mutations were in advanced disease stages. These findings suggest that mutational inactivation of the p53 gene is infrequent but is involved in the tumorigenesis of several types of haematologic neoplasms at least in some cases. 相似文献
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目的:分析我国重庆地区肝细胞癌P53基因失活机制及突变谱。方法:采用PCR—RFLPPCRSSCP和PCR直接测序技术对来自我国重庆地区28例肝细胞癌P53抑癌基因结构异常进行了分析。结果:61.51%的肝癌存在p53的杂合缺失:50%肝癌伴有p53基因突变,其突变模式为突普通散在于567和8外显子,其中第7外显子249们密友情子突弯率最高(21%);具有突变的肝癌多同时伴夺缺失。伴有p53基因结构异常的肝癌均属进展期。结论:我国重庆地区肝癌存在P53基因结构异常,P53基因结构异常,p53基因突变模式反映了该地区肝癌发生可能与肝炎病互和黄贡互素两种因素及其相互作用有关;p53基因结构异常属肝癌晚期事件,可能参加与肝癌的进展过程。 相似文献
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肿瘤抑制基因p53及p16与胃癌生物学行为的关系 总被引:1,自引:4,他引:1
目的探讨肿瘤抑制基因p53和p16异常与胃癌生物学行为的关系.方法采用免疫组化ABC法检测58例原发性胃癌(男38例,女20例,年龄37岁~76岁),P53和P16蛋白的表达变化.所有组织均新鲜取材,并迅速用850ml/L酒精固定,石蜡包埋,连续切片.结果受检组织中p53和p16阳性表达率分别为517%(30/58)和483%(28/58).P53蛋白在低分化胃癌(700%)、进展期胃癌(569%)、淋巴结阳性胃癌(741%)中的表达率高于相应的高分化、早期、淋巴结阴性胃癌的表达率(273%,143%,323%)(P<005),且p53高表达多见于弥散型胃癌(同肠型胃癌比)、累及浆膜的胃癌也较局限于粘膜层的胃癌有更高的P53蛋白表达(P<005);P16蛋白表达与胃癌大多数生物学行为无明显关系,但其在淋巴结阳性胃癌中的表达率(333%),低于淋巴结阴性胃癌中的表达率(613%);相关性分析显示;p53阳性组织大多伴有P16蛋白阳性表达(P<005).结论P53蛋白异常表达对胃癌生物学行为有广泛影响,P16蛋白表达缺失可能是胃癌淋巴结转移的重要促发因素. 相似文献
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Immunohistochemical demonstration of the p53 tumour suppressor gene product in cancer of the pancreas and chronic pancreatitis 总被引:5,自引:0,他引:5
C. SOON LEE MOIRA RUSH DORA CHARALAMBOUS JÜRGEN RODE 《Journal of gastroenterology and hepatology》1993,8(5):465-469
The p53 tumour suppressor gene and its protein products after point mutations are currently attracting wide attention in the investigation of human tumours. In this study we present the findings on percutaneous pancreatic biopsies of 82 cases after routine processing and immunostaining for the polyclonal p53 antibody CM1, an antibody directed against both wild and mutant forms of p53 protein. There were 51 carcinomas, 5 islet cell tumours, 16 cases of chronic pancreatitis (including one with atypical ductal epithelium) and seven histologically normal pancreatic biopsy specimens. None of the seven normal cases showed any definite nuclear immunostaining for p53. Thirty-two (63%) of the pancreatic adenocarcinomas showed moderate to intense immunoreactivity. Of the 16 cases of chronic pancreatitis, 11 were negative and three showed equivocal immunostaining. The one case with ductal epithelial atypia showed mild to moderate immunoreactivity. All islet cell tumours were negative. The expression of the p53 gene, therefore, appears increased in the majority of pancreatic adenocarcinomas while this is not observed in chronic pancreatitis or normal pancreatic tissue. Nuclear immunoreactivity for p53 protein may represent mutant forms because of the short half-life of the wild-type protein. The lack of p53 expression in some cases of pancreatic adenocarcinoma may be due to different types of mutant proteins not detectable by the CM1 antibody. Nuclear immunoreactivity to the p53 protein in pancreatic biopsy is more suggestive of a malignant tumour than chronic pancreatitis. 相似文献
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Mutated p53 protein expression and proliferative activity in advanced gastric cancer. 总被引:6,自引:0,他引:6
M Ikeguchi H Saito A Kondo S Tsujitani M Maeta N Kaibara 《Hepato-gastroenterology》1999,46(28):2648-2653
BACKGROUND/AIMS: When the DNA of cells is damaged, wild-type p53 protein induces the expression of p21 (Waf1/Cip1/Sdi1), and regulates the progression of the cell cycle by inducing G1 arrest. Thus, wild type p53 or p21 protein negatively regulates cancer cell proliferation. However, in tumor with loss of expression of functional wild-type p53 protein by mutation or allelic deletion of the gene for p53, whether the proliferative activity of cancer cells might be accelerated or not is unclear. In this study, we investigated the correlation between the level of expression of mutated p53 protein and the proliferative activity of cancer cells in advanced gastric cancer. METHODOLOGY: Ninety-seven samples from patients with gastric cancer that had invaded the serosa without lymph node metastasis (t3, n0, stage II) were investigated by immunohistochemical staining with a monoclonal antibody against p53 and against p21, and with the monoclonal antibody Ki-67. DNA ploidy patterns were analyzed by flow cytometry. The immunoreactivity against p53 and the proliferative activity of cancer cells were scored in terms of a labeling index (LI; percentage of immunostained cells) in each case. Moreover, the prognostic values for 93 surviving patients were evaluated by univariate and multivariate analysis. RESULTS: The mean p53 LI was 24% (range: 0-82.4%) and the mean Ki-67 LI was 23.1% (range: 0-70.7%) in 97 tumors. The expression of p21 protein was detected in 30 of 97 tumors (30.9%) and DNA aneuploidy was detected in 36 of 97 tumors (37.1%). There was significant correlation between the p53 LI and the Ki-67 LI (r = 0.61, t = 7.456, p < 0.001) in 97 tumors. Although, no significant difference was detected, the mean p53 LI (18.3%) of 30 tumors with expression of p21 protein was lower than that of 67 tumors without expression of p21 protein (26.6%, p = 0.096). However, no significant correlation between expression of p21 protein and Ki-67 LI was observed. The p53 LI was not an independent prognostic factor in 93 surviving patients by multivariate survival analysis (p = 0.069). However, the 5-year survival rate of 50 patients with a low level of p53 LI (p53 LI (< or = 10%, 78.3%) was significantly better than that of 43 patients with a high level of p53 LI (p53 LI > 10%, 62.1%, p = 0.045). CONCLUSIONS: Accumulation of mutated p53 protein might suppress the expression of p21 protein in gastric adenocarcinoma, and cancer cells with overexpression of mutated p53 protein might have a high proliferative activity. 相似文献
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Analysis of p53 tumour suppressor gene somatic mutations in rheumatoid arthritis synovium 总被引:5,自引:1,他引:5
Inazuka M Tahira T Horiuchi T Harashima S Sawabe T Kondo M Miyahara H Hayashi K 《Rheumatology (Oxford, England)》2000,39(3):262-266
OBJECTIVE: In order to study the role of the p53 tumour suppressor gene in the proliferation of rheumatoid arthritis (RA) synovium, we analysed the mutation of p53 in the synovial fibroblast-like type B synoviocyte from RA patients. METHODS: Synovial fibroblast-like type B synoviocytes were prepared from the synovial tissues from nine Japanese patients with RA. The p53 cDNA region from exons 4-11 was screened for mutations by the streamlined mutation detection method in which polymerase chain reaction (PCR) products are post-labelled and are analysed by automated capillary electrophoresis using single-strand conformation polymorphism conditions, followed by direct sequencing of the subclones of the PCR products. RESULTS: p53 mutation with possible functional alteration was detected in four of the nine RA patients (44.4%). Of a total of 262 p53 cDNA subclones, 10 subclones were carrying 10 p53 mutations, eight of which were associated with amino acid alterations or protein truncation. Of the p53 functional mutations, a substitution of Gly at amino acid residue 245 to Asp (G245D) was identified in two patients in three subclones. G245D was the first mutation that was recurrently identified in different RA individuals. G245D is also one of the relatively common mutations in human cancers. CONCLUSIONS: In some patients with RA, dysfunction of p53 might play a role in the proliferation of the synovial tissue. G245D mutation might especially need further study as it is the first recurrently identified p53 mutation in RA and is also one of the frequently identified mutations in human cancers. 相似文献
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Tidow H Melero R Mylonas E Freund SM Grossmann JG Carazo JM Svergun DI Valle M Fersht AR 《Proceedings of the National Academy of Sciences of the United States of America》2007,104(30):12324-12329
The homotetrameric tumor suppressor p53 consists of folded core and tetramerization domains, linked and flanked by intrinsically disordered segments that impede structure analysis by x-ray crystallography and NMR. Here, we solved the quaternary structure of human p53 in solution by a combination of small-angle x-ray scattering, which defined its shape, and NMR, which identified the core domain interfaces and showed that the folded domains had the same structure in the intact protein as in fragments. We combined the solution data with electron microscopy on immobilized samples that provided medium resolution 3D maps. Ab initio and rigid body modeling of scattering data revealed an elongated cross-shaped structure with a pair of loosely coupled core domain dimers at the ends, which are accessible for binding to DNA and partner proteins. The core domains in that open conformation closed around a specific DNA response element to form a compact complex whose structure was independently determined by electron microscopy. The structure of the DNA complex is consistent with that of the complex of four separate core domains and response element fragments solved by x-ray crystallography and contacts identified by NMR. Electron microscopy on the conformationally mobile, unbound p53 selected a minor compact conformation, which resembled the closed conformation, from the ensemble of predominantly open conformations. A multipronged structural approach could be generally useful for the structural characterization of the rapidly growing number of multidomain proteins with intrinsically disordered regions. 相似文献
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原发性胃癌p53基因突变 总被引:2,自引:0,他引:2
李中信 《World journal of gastroenterology : WJG》1996,2(1):41-43
目的 p53基因是当前抑癌基因研究中的热点之一。迄今,有关 p53基因异常与胃癌临床病理学参数如大体类型、临床分期、组织分化程度,浸润深度及淋巴结转移之间的关系尚无定论。Tumura 报告p53基因改变主要发生于异倍体瘤,国内尚无报道。本实验目的主要是分析中国人原发性胃癌 p53基因突变与这些病理参数,包括 DNA 倍体之间的关系。方法用聚合酶链式反应—单构象多态分析(PCR—SSCP)技术对20例原发性胃癌 p53基因外显子5—8突变进行检测。结果 8例(40%)发生了突变,其中2例发生在外显子7,4例发生在外显子8。0至Ⅲ期均有突变存在。66.7%(6/9)的异倍体瘤检测到了p53突变,而二倍体瘤中只有18.2%(2/11)发生了 p53突变。结论 p53基因突变与胃癌临床病理参数如大体类型、分期、组织分化程度、浸润深度及淋巴结转移之间无明显关系,而与胃癌 DNA 倍体改变有关。 相似文献
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Dual role of serum soluble E-cadherin as a biological marker of metastatic development in gastric cancer 总被引:7,自引:0,他引:7
Juhasz M Ebert MP Schulz HU Röcken C Molnar B Tulassay Z Malfertheiner P 《Scandinavian journal of gastroenterology》2003,38(8):850-855
BACKGROUND: Soluble E-cadherin serum levels as a potential biological marker for gastric cancer were analysed with special consideration to clinical and pathological features. METHODS: Seventy-one healthy control subjects and 166 patients with gastric cancer were enrolled. Gastric cancer patients were classified into intestinal-type (51%) and diffuse-type (49%), according to Laurén. Soluble E-cadherin serum levels were measured with enzyme-linked immunosorbent assay. RESULTS: The mean logarithmic concentrations of soluble E-cadherin in gastric cancer patients were significantly higher than those of control subjects, with an average of 4.03 (+/- 0.32) versus 3.86 (+/- 0.24), respectively (P < 0.0001). The concentration of soluble E-cadherin was significantly higher in the intestinal-type group than in the diffuse-type group, with an average of 4.07 +/- 0.3 versus 3.98 +/- 0.34, respectively (P = 0.0494). In the intestinal-type group, concentrations of soluble E-cadherin were significantly higher in more advanced stages (stages III-IV) than in earlier stages (stages I-II), with an average of 4.13 +/- 0.29 versus 3.96 +/- 0.31, respectively (P = 0.0234). In the diffuse-type group, concentrations of soluble E-cadherin were significantly higher in localized than in metastatic gastric cancer, with an average soluble E-cadherin concentation of 4.15 +/- 0.3 versus 3.95 +/- 0.32, respectively (P = 0.0139). CONCLUSION: Serum soluble E-cadherin concentrations exhibit a completely different pattern in intestinal-type and diffuse-typegastric cancer. Serum levels are increased in intestinal-type gastric cancer, especially in advanced stages, whereas in diffuse-type gastric cancer E-cadherin levels are decreased in advanced, metastasized cancer.We conclude that soluble E-cadherin concentrations should be interpreted along with Laurén classification and thus might serve as a biological marker in intestinal-type gastric cancer. 相似文献
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p53与胃癌研究进展 总被引:2,自引:0,他引:2
p53基因突变与人类很多肿瘤有关,在正常细胞及组织中,p53蛋白的半衰期很短,浓度很低,不能用免疫组化(immunohistochemistry,IHC)法测出。相反,肿瘤中有突变体p53蛋白,其稳定性增强,量积聚,可用免疫组化法测出,一般认为,胃癌组织中p53的积聚是恶性程度高的表现,用免疫组化法测定p53,结合其它常规的检查,可用来更好地设计合理的治疗方案。 相似文献
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David G. McFadden Amanda Vernon Philip M. Santiago Raul Martinez-McFaline Arjun Bhutkar Denise M. Crowley Martin McMahon Peter M. Sadow Tyler Jacks 《Proceedings of the National Academy of Sciences of the United States of America》2014,111(16):E1600-E1609
Anaplastic thyroid carcinoma (ATC) has among the worst prognoses of any solid malignancy. The low incidence of the disease has in part precluded systematic clinical trials and tissue collection, and there has been little progress in developing effective therapies. v-raf murine sarcoma viral oncogene homolog B (BRAF) and tumor protein p53 (TP53) mutations cooccur in a high proportion of ATCs, particularly those associated with a precursor papillary thyroid carcinoma (PTC). To develop an adult-onset model of BRAF-mutant ATC, we generated a thyroid-specific CreER transgenic mouse. We used a Cre-regulated BrafV600E mouse and a conditional Trp53 allelic series to demonstrate that p53 constrains progression from PTC to ATC. Gene expression and immunohistochemical analyses of murine tumors identified the cardinal features of human ATC including loss of differentiation, local invasion, distant metastasis, and rapid lethality. We used small-animal ultrasound imaging to monitor autochthonous tumors and showed that treatment with the selective BRAF inhibitor PLX4720 improved survival but did not lead to tumor regression or suppress signaling through the MAPK pathway. The combination of PLX4720 and the mapk/Erk kinase (MEK) inhibitor PD0325901 more completely suppressed MAPK pathway activation in mouse and human ATC cell lines and improved the structural response and survival of ATC-bearing animals. This model expands the limited repertoire of autochthonous models of clinically aggressive thyroid cancer, and these data suggest that small-molecule MAPK pathway inhibitors hold clinical promise in the treatment of advanced thyroid carcinoma.Mutations in the v-raf murine sarcoma viral oncogene homolog B (BRAF) kinase occur in ∼60% of papillary thyroid carcinomas (PTCs) (www.cbioportal.org/public-portal/data_sets.jsp). PTC generally exhibits an excellent prognosis with conventional therapy, including surgery and selective use of radioiodine (1). PTC may progress to clinically aggressive forms of thyroid cancer, including poorly differentiated thyroid carcinoma (PDTC), which exhibits more rapid growth and poorer clinical outcome. Less commonly, PTC progresses to undifferentiated (anaplastic) thyroid carcinoma (ATC) that is associated with a grim prognosis with a median survival of 5 mo and a 1-y survival of only 20% (2).Focused sequencing of clinically aggressive subsets of thyroid cancers including PDTC and ATC suggests acquired cooperating mutations drive thyroid cancer progression (3, 4). Mutations in tumor protein p53 (TP53) occur with increasing frequency in more aggressive forms of thyroid cancer, culminating in ATC, which harbors the highest frequency of TP53 mutations (5–7). ATC may progress from well-differentiated thyroid carcinomas and is also believed to arise spontaneously, possibly from clinically undetectable microscopic well-differentiated thyroid tumors. In the former scenario, ATCs frequently harbor mutations in BRAF, and these mutations are concordant between the anaplastic and papillary components. This implicates BRAF mutation as an initiating somatic genetic event and supports the hypothesis that loss of p53 function is important for progression to ATC (3, 8).Mouse models of thyroid cancer have supported the model of acquired mutations driving tumor progression. Although each study has technical limitations, including embryonic oncogene expression and/or elevated circulating thyroid-stimulating hormone (TSH) levels, this work generally supports the notion that BRAFT1799A is sufficient to initiate PTC (9–12). In addition, deletion of p53 enabled tumor progression to high-grade thyroid carcinomas in a transgenic mouse model of translocations targeting the ret proto-oncogene (RET/PTC) driven PTC, and a model of follicular thyroid carcinoma initiated by tissue-specific phosphatase and tensin homolog (Pten) deletion (13, 14). These studies provide functional evidence of an important tumor suppressive role for p53 during thyroid carcinoma progression, although to date this has not been tested in models of BRAF-mutant PTC.Given the high frequency of BRAF and RAS mutations in thyroid carcinomas and the success of targeted therapy trials for advanced thyroid cancers, the potential utility of small-molecule inhibitors of the MAPK pathway has garnered much recent attention (15). These drugs have also been studied in models of BRAF-mutant thyroid carcinoma. Initial observations using a thyroid-specific doxycycline-inducible BRAFT1799A allele suggested that BRAF or mapk/Erk kinase (MEK) inhibition induced thyroid carcinoma regression and differentiation (9). However, a recent study from the same laboratory showed a mitigated response to BRAF (PLX4032, vemurafenib) inhibition in human papillary and ATC cell lines and in an endogenous BrafV600E-driven PTC mouse model. In response to PLX4032/vemurafenib, feedback inhibition of the human epidermal growth factor receptor 3 (HER3) receptor tyrosine kinase was abrogated, leading to reactivation of MAPK signaling (16). In addition, responses in patients treated with the BRAF inhibitor vemurafenib have exhibited modest activity (17).To develop an adult-onset autochthonous model of clinically aggressive thyroid carcinoma, we generate a thyroid-specific CreER transgenic mouse and use conditional BrafT1799A and Trp53 alleles. We demonstrate that expression of BRAFV600E is sufficient to initiate tumorigenesis in adult animals, and p53 loss enables progression to bona fide ATC recapitulating the cardinal features of the human disease including intrinsic resistance to BRAF inhibitors. 相似文献
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AIM: To investigate the role of the polymorphism of p53 codon 72 in early gastric cancer (EGC) and advanced gastric cancer (AGC) in Korean patients.METHODS:DNA was extracted from blood samples of gastric cancer patients (n = 291) and controls (n=216).tn the p53 codon 72 genotypes were determined by PCR-RFLP.RESULTS: Patients with gastric cancer had a significantly higher frequency of the homozygous proline (Pro) allele than the control (P=0.032). Patients with AGC had a significantly higher frequency of the Arg/Arg (arginine)allele (P=0.038) than EGC and a similar Pro/Pro allele.The signet ring cell type had a higher frequency of the Pro/Pro allele than other types (P=0.031). The Pro/Pro genotype carries a 3.9-fold increased risk of developing gastric cancer (95% CI,1.3-15.4,P=0.039)when compared to Arg/Arg and Arg/Pro genotypes and to develop EGC is a 5.25 fold increased risk (95% CI,1.8-19.6,P=0.021).CONCLUSION: The Pro/Pro genotype of the p53 codon 72 polymorphism carries a higher risk for gastric cancer in general and is also associated with a much higher risk for EGC than AGC. 相似文献