胃粘膜中增殖细胞核抗原及p53、K-ras、bc1-2基因蛋白的表达

为了解胃粘膜癌变过程中多基因蛋白的表达及意义，应用免疫组化技术检测１０７例内镜活检胃粘膜组织中增殖细胞核抗原（ＰＣＮＡ）及ｐ５３、Ｋ－ｒａｓ、ｂｃ１－２基因蛋白。结果各基因蛋白阳性率随病变程度加重而增加。Ⅲ型肠化生者增殖细胞核抗原标记指数（ＰＣＮＡＬＩ）及Ｋ－ｒａｓ、ｂｃ１－２基因蛋白阳性率较之Ⅰ、Ⅱ型肠化亦有增高趋势但未见统计学差异（Ｐ≥０．０５）。１２例Ⅲ型肠化中有９例出现Ｋ－ｒａｓ或ｂｃ１－２蛋白异常表达（７５．０％），显著高于Ⅰ、Ⅱ型肠化（２３．８％）（Ｐ＜０．０５）。在肠型及胃型胃癌ｐ５３表达率为５３．３％、３７．０％（Ｐ＞０．０５）；Ｋ－ｒａｓ为４０．０％、７．４％（Ｐ＜０．０５）；ｂｃ１－２为９３．３％、５９．３％（Ｐ＜０．０５）。１５例肠型胃癌中有９例同时存在２种或２种以上蛋白异常表达（３例ｐ５３与ｂｃ１－２表达异常，２例ｂｃ１－２与Ｋ－ｒａｓ表达异常，４例ｐ５３与ｂｃ１－２、Ｋ－ｒａｓ异常表达），高于胃型胃癌（Ｐ＜０．０５）。结果提示以上基因产物的表达异常与胃粘膜癌变相关，Ⅲ型肠化属于胃癌癌前病变范畴，肠型胃癌发生发展的分子机制与胃型胃癌显著不同。     

ras癌基因第12密码子点突变和胃癌患者预后关系的研究

对福尔马林固定，石蜡包埋的胃癌组织使用移聚酶链式反应－限制性片段长度多态性（简称ＰＣＲ－ＲＦＬＰ）技术同时进行ｃ－Ｈａ－ｒａｓ基因第１２位和６１位，Ｎ－ｒａｓ基因第１２位，ｋ－ｒａｓ第１２位和１３位密码子点突变的研究，发现３３．３％（１４／４２），的胃癌有ｃ－Ｈａ－ｒａｓ基因第１２位密码子的点突变；４．８％（２／４２）的病例有ｋ－ｒａｓ基因第１２位密码子的点突变，点突变的发生与患者的预后，淋巴结转     

抗基因及反义寡脱氧核苷酸抑制N－ras表达及对肝癌细胞增殖的影响

探讨抗基因及反义寡核苷酸对肝癌的潜在治疗作用。方法合成了针对Ｎ－ｒａｓ转录、翻译起始区的１２ｍｅｒ硫代磷酸抗基因寡脱氧核苷酸（Ｓ－ＯＤＨａｎ）、１９ｍｅｒ硫代磷酸反义寡脱氧核苷酸（Ｓ－ＯＤＮＡａｓ）。采用ＥＬＩＳＡ、斑点杂交法分别检测了寡脱氧核苷酸处理的肝癌ＢＥＬ７４０２细胞内ｐ２１、Ｎ－ｒａｓｍＲＮＡ水平，观察了对ＢＥＬ７４０２肝癌细胞生长的影响。结果处理细胞内Ｎ－ｒａｓｍＲＮＡ水平低于对照组，以Ｓ－ＯＤＮａｎ组更明显。两者对ｐ２１合成的抑制率达８０％，６７．５％。两者的细胞生长抑制率达８８％、７７％，并呈剂量依赖性。两者处理的细胞３Ｈ－ＴｄＲ掺入降低为对照组的２１．８％、３０．５５％；ＡＦＰ水平明显低于对照组（Ｐ直＜０．００１）。结论Ｓ－ＯＤＮａｎ．Ｓ－ＤＤＮａｓ能有效抑制Ｎ－ｒａｓ表达及相关肝癌细胞增殖；也进一步说明Ｎ－ｒａｓ在肝癌发生中起作重要作用。     

胃粘膜中增殖细胞核抗原及p53,K—ras,bcl—2基因蛋白的表达

为了解胃粘膜癌变过程中多基因蛋白的表达及意义，应用免疫组化技术检测１０７例内镜活检胃粘膜组织中增殖细胞核抗原（ＰＣＮＡ）及ｐ５３、Ｋ－ｒａｓ，ｂｃｌ－２基因蛋白。结果各基因蛋白阳性率随病变程度加重而增加。Ⅲ型肠化生者增殖细胞核抗原标记指数（ＰＣＮＡ ＬＩ）及Ｋ－ｒａｓ、ｂｃｌ－２基因蛋白阳性率较之Ⅰ、Ⅱ型肠化亦有增高趋势但未见统计学差异（Ｐ≥０．０５）。１２例Ⅲ型肠化中有９例出现Ｋ－ｒａｓ和ｂｃｌ     

萎缩性胃炎及异型增生ras p21的表达

萎缩性胃炎及异型增生ｒａｓｐ２１的表达倪桂宝我们应用ＡＢＣ免疫组化法进行了ｒａｓ癌基因产物ｐ２１单克隆抗体检测，旨在探讨ｒａｓ癌基因在胃癌前状态中的表达及其与胃癌发生的关系。一、材料和方法材料选自本院１９８８～１９９３年间胃镜活检标本，其中慢性萎缩性...     

胃癌中癌基因ras c-myc mRNA表达的临床意义

目的探索癌基因ｒａｓ和ｃｍｙｃｍＲＮＡ在胃癌发生中单独和协同作用关系，选择胃癌的早期诊断依据．方法应用原位分子杂交（ＩＳＨ）技术，检测８８例胃癌组织中Ｈｒａｓ，Ｋｒａｓ，ＣｍｙｃｍＲＮＡ的表达，结合临床资料综合分析．结果Ｋｒａｓ，Ｈｒａｓ，ＣｍｙｃｍＲＮＡ在肿瘤区的阳性表达分别为７８４％，７０５％和５８０％，在癌旁区的阳性表达分别为２８４％，２７３％和２０５％，呈梯度下降趋势，３种癌基因的表达与胃癌的临床分型、组织学分类及有无淋巴结转移无关，ＣｍｙｃｍＲＮＡ的表达与患者生存期关系密切（Ｐ＜００５）．结论Ｋｒａｓ，Ｈｒａｓ，ＣｍｙｃｍＲＮＡ的表达可作为胃癌早期诊断的参考依据．     

幽门螺杆菌阳性胃粘膜病变AgNOR和rasp21的表达

目的研究幽门螺杆菌阳性（Ｈｐ＋）不同胃粘膜病变的ＡｇＮＯＲ数量及ｒａｓｐ２１阳性表达率，探讨其生物学行为及Ｈｐ在此过程中可能的作用．方法经内窥镜病理检查证实胃粘膜病变（慢性浅表性胃炎、慢性萎缩性胃炎、肠上皮化生、异型增生、胃癌）共计２７８例．通过Ｈｐ检测（ＣＬＯｔｅｓｔ结合ＷａｒｔｈｉｎＳｔａｒｙ染色）证实其中１４６例阳性，１３２例阴性．分别对其粘膜标本作了ＡｇＮＯＲ定量及ｒａｓｐ２１表达的研究，比较不同胃粘膜病变中Ｈｐ阳性和阴性组间ＡｇＮＯＲ数目和ｒａｓｐ２１阳性表达率．结果除慢性浅表性胃炎外各病变中Ｈｐ＋组的ＡｇＮＯＲ数量均显著高于Ｈｐ－组（Ｐ＜００５或Ｐ＜００１）；除慢性浅表性胃炎及慢性萎缩性胃炎外各病变中Ｈｐ＋组的ｒａｓｐ２１阳性表达率均显著高于Ｈｐ－组（Ｐ＜００５）．结论Ｈｐ＋胃粘膜病变具有更多的肿瘤生物学行为，该菌可能刺激胃上皮细胞的过度增殖而启动恶性变．     

DNA甲基化与肝癌研究进展

ＤＮＡ甲基化是转录水平的ＤＮＡ的修饰方式之一,其在调节基因表达及维持细胞正常分化中起着重要作用。正常情况下细胞内约７５％的胞嘧啶一鸟嘌呤碱基对中的胞嘧啶（Ｃ）处于甲基化状志,它是维持细胞稳定性的重要因素,其水平下降或模式改变可导致基因结构和功能的异常,是细胞癌变中重要的一步。大量实验表明,在肿瘤的发生发展中存在ＤＮＡ甲基化水平和模式的紊乱。研究发现,化学致癌剂诱发的大鼠肝结节增生和肝癌中可发现ｃ－ｍｙｃ、ｃ－Ｈａ－ｒａｓ基因的ＤＮＡ甲基化模式改变,而相应癌基因表达增加［１］。我国为肝癌（ＨＣＣ）…     

用PCR和直接测序方法分析胰腺的c-Ki-ras基因点突变

用ＰＣＲ和直接测序方法分析胰腺的ｃ－Ｋｉ－ｒａｓ基因点突变徐永泉，刘述信，夏玉亭人类肿瘤中最常见的与转化活性有关的基因ｒａｓ家族癌基因（Ｎ－ｒａｓ，Ｋｉ－ｒａｓ，Ｈａ－ｒａｓ）。ｒａｓ基因的激活在肿瘤发生中起重要作用。ｒａｓ基因激活的主要方式是点突变...     

肝细胞癌患者HCV感染及其致癌机理研究

目的：揭示肝细胞癌患者的ＨＣＶ感染情况及其致癌机理。方法：应用免疫组织化学ＳＰ法对４６例肝细胞癌癌组织及其３８例癌旁肝组织中丙型肝炎病毒核心抗原作了定位研究，并进一步检测其中癌基因ｒａｓ、ｃｍｙｃ和抑癌基因ｐ５３的蛋白表达。结果：核心抗原的检出率分别为２１７％和３６８％，４６例肝癌患者中１６例存在丙型肝炎病毒感染；ｐ２１、ｃｍｙｃ和ｐ５３蛋白在癌组织中的检出率分别为５８７％、６７４％和３０４％；丙型肝炎病毒感染的阳性与阴性组患者之间的ｐ２１、ｃｍｙｃ和ｐ５３蛋白的表达无显著性差异（Ｐ＞０．０５）；两组患者癌旁肝硬化的阳性率分别为８７５％和８６７％（Ｐ＞００５）。结论：上述结果提示丙型肝炎病毒致肝细胞癌的机理可能与ｒａｓ、ｃｍｙｃ和ｐ５３基因的激活或失活无关，其机理尚待进一步探索     

Frequent overexpression, but not activation by point mutation, of ras genes in primary human gastric cancers

To define the extent of involvement of ras oncogenes in human gastric cancers, we surveyed for the presence of ras oncogenes, activated by either point mutations within their coding sequences or overexpression of ras protein p21, by the combined use of several analytic techniques. Primary gastric cancers were first analyzed by deoxyribonucleic acid transfection assay using NIH/3T3 cells as recipients and by restriction enzyme analysis, which detects point mutations at codon 12 of the H-ras gene. None of seven tumors analyzed scored as positive. Furthermore, none of them had ras p21 with altered electrophoretic mobility on immunoprecipitation and Western blotting, confirming the absence of ras oncogenes activated by point mutations in these tumors. However, in 6 of 7 tumors, the amounts of p21 exceeded that in human placenta. Amplification of the K-ras gene was found in 1 of 11 (including the 7 described above) gastric cancers. Immunohistochemical analysis of ras p21 expression in these 11 tumors was then carried out using the anti-ras p21 monoclonal antibody RAP-5. All cancers showed more reactivity with RAP-5 than did normal mucosa adjacent to the cancers, indicating increased expression of ras p21. These results indicated that transformation of the stomach mucosa from the normal to the malignant phenotype is rarely associated with activation of ras genes by point mutations, but is frequently associated with enhanced expression of ras p21.     

胃幽门螺杆菌与胃癌发病机制关系的分子生物学研究

近期流行病学研究已发现胃幽门螺杆菌(HP)是胃癌发生的重要始发因素。本文应用敏感特异的聚合酶链反应(PCR)方法检测胃粘膜组织HP,分析其与C—Ha—ras第12位密码子点突变,ras基因产物P~(21)蛋白表达及DNA倍体的关系。发现C—Ha—ras点突变率于HP阳性组高于阴性组,并且HP阳性组较HP阴性组出现rasP~(21)表达增强,说明HP感染与C—Ha—ras基因活化有关,HP感染后DNA含量及S％期细胞明显增高,提示核苷酸代谢旺盛,DNA损伤及非整倍体的危险性也增高。本研究为HP致胃癌的可能机制从分子水平及细胞代谢水平作了初步研究。     

胃癌、胃黏膜不典型增生和胃炎组织中p185和p21蛋白的表达及其临床意义

目的研究胃癌、胃黏膜不典型增生和胃炎组织中c-erbB-2和ras癌基因产物p185和p21蛋白的表达,以探讨p185和p21蛋白在胃癌发生、发展过程中的作用。方法采用免疫组化S-P法检测39例胃癌组织、24例胃黏膜不典型增生组织和33例胃炎组织p185和p21蛋白的表达。结果p185蛋白在慢性胃炎、轻度胃黏膜不典型增生组织中不表达;在中、重度胃黏膜不典型增生组织中的表达率分别为11.8%和66.7%,差异有显著性(P<0.05),表达程度明显增强(P<0.05);在高、中、低分化胃癌组织中的表达率分别为77.8%、50.0%和29.2%,差异有显著性(P<0.05),高分化胃癌组织中p185蛋白的表达程度明显高于中、低分化胃癌组(P<0.05)。p21蛋白在胃黏膜不典型增生及胃癌组织中的表达率和表达程度较慢性胃炎组明显增高,差异有显著性(P<0.01);在轻、中、重度胃黏膜不典型增生组织中的表达率分别为25.0%、82.4%和100.0%,差异有显著性(P<0.05),表达程度明显增强(P<0.05);在高、中、低分化胃癌组织中的表达率分别为44.4%、66.7%和95.8%,差异有显著性(P<0.01),表达程度明显增强(P<0.05)。p185和p21蛋白在胃癌组织中的表达呈负相关(r=-0.450,P<0.01)。结论c-erbB-2和ras癌基因产物p185和p21蛋白在胃癌致病机制中起重要作用,p21蛋白作用于胃癌发生的早期阶段,p185蛋白可能作用于胃癌发生的较晚期。     

Clinical significance of ras p21 overexpression for patients with an advanced colorectal cancer

The expression of ras oncogene product p21 was examined in 45 paraffin-embedded sections of primary advanced colorectal cancers, using the anti-v-H-ras p21 monoclonal antibody Y13-259. Fourteen of these specimens (31 percent) were stained positively. The incidence of lymphatic vessel invasion of cancer cells and lymph node metastasis correlated statistically with the overexpression of ras p21. The depth of invasion and incidence of liver metastasis in the p21-positive group were more prominent than in the p21-negative group. Statistically significant differences were evident in operative curability and clinical stage at initial surgery and in the longterm survival rate between these groups (P <0.05). We propose that ras p21 overexpression may serve as a marker to predict the prognosis of colorectal cancer.     

Cellular analysis of c-Ha-ras gene expression in rat liver after CCl4 administration

Expression of the c-Ha-ras proto-oncogene is specifically enhanced during liver regeneration, in parallel with increased DNA replication, which suggests that c-Ha-ras may play a role in the control of regeneration. In this study, an in situ hybridization technique was applied for analysis of expression of the c-Ha-ras gene at the cellular level during liver regeneration induced by CCl4 administration. The in situ hybridization was compared with the observation for the p21c-Ha-ras protein, the corresponding protein of the c-Ha-ras gene, by immunohistochemistry. In normal rat liver, a few hepatocytes expressed the mRNAs and the corresponding proteins without any preferential localization. Zonal heterogeneity of c-Ha-ras gene expression first became evident at 12 hr after CCl4 administration, a higher number of gene products being detected in the pericentral zone than in the periportal zone. This heterogeneity became maximal at 24 hr after CCl4 administration. Zonal heterogeneity in the level of the p21c-Ha-ras protein paralleled that in the level of gene expression. Furthermore, both hepatocytes and nonparenchymal cells participated in expression of the c-Ha-ras gene during liver regeneration.     

Helicobacter pylori Infection and Oncogene Expressions in Gastric Carcinoma and Its Precursor Lesions

Although it is fairly well accepted that Helicobacter pylori infection plays a significant role in causing gastric cancer, the exact mechanisms involved in its pathogenesis are unclear. We have examined the relationship between H. pylori infection and oncogene expression in different stages of disease progression from precursor lesions to gastric carcinoma. We used Diff-Quik stain to diagnose H. pylori infection and immunohistochemical stains against c-erbB-2, p53, ras, c-myc, and bcl-2 to determine expression of oncogenes. H. pylori infection was found in all cases of chronic gastritis, atrophic gastritis, intestinal metaplasia, and early gastric carcinoma, and in 16 of 30 (53%) cases of advanced gastric carcinoma. Overexpression of c-erbB-2 was found in 2 (7%) cases of advanced gastric carcinoma, which were H. pylori negative. Suppressor gene, p53, was overexpressed in 3 (30%) cases of intestinal metaplasia, 2 (33%) cases of early gastric carcinoma, and 18 (60%) cases of advanced gastric carcinoma. Of these 18 p53-positive advanced gastric cancer cases, 11 (61%) were H. pylori positive. Expression of ras p21 was found in 4 (40%) cases of H. pylori-negative normal mucosa, 10 (100%) cases of chronic gastritis, 1 (10%) case of atrophic mucosa, 6 (60%) cases of intestinal metaplasia, 2 (33%) cases of nonneoplastic mucosa adjacent to early gastric carcinoma, and 7 (23%) nonneoplastic mucosa adjacent to advanced gastric carcinoma, all of which showed H. pylori. No evidence of expression of either c-myc or bcl-2 was detected in any of the above-mentioned samples. The data suggest that H. pylori infection may increase expression of ras p21 proteins and induce p53 suppressor gene mutation early in the process of gastric carcinogenesis.     

胃复春对胃癌癌前病变的治疗及对p21ras、p53表达的调节作用研究

目的：研究胃复春片对胃黏膜中、重度异型增生的治疗作用及对P21^ras、P53蛋白表达的调节作用，探讨其逆转胃癌癌前病变的治疗价值及机制。方法：内镜下病理活检证实为中、重度异型增生患者共58例，随机分为胃复春治疗组32例，维酶素对照组26例。治疗前后比较临床疗效，镜下改变，病理改变及胃黏膜固定部位活检标本P21^ras、P53蛋白S-P法免疫组化染色变化情况。结果：胃复春组临床症状的总有效率81．25％，内镜下改变总有效率71．88％，病理学改变总有效率65．63％，均优于维酶素对照组(分为53．85％、46．15％、34．61％)。P21^ras、P53蛋白在中、重度异型增生组织中有过度表达，胃复春能使其表达明显减弱。结论：胃复春对胃黏膜中、重度异型增生有良好的治疗作用，能促进病变胃黏膜的逆转，并对P21^ras、P53蛋白的表达有一定的调节作用，降低癌变危险性，可用于胃癌癌前病变的治疗。     

Activating mutations of the c-Ha-ras protooncogene in chemically induced hepatomas of the male B6C3 F1 mouse.

Activated c-Ha-ras protooncogenes have recently been identified in the DNA of some spontaneous hepatic tumors found in 2-year-old B6C3 F1 mice. Activation of c-Ha-ras has now been demonstrated in DNA from well-differentiated hepatomas initiated by a single dose of carcinogen given to male B6C3 F1 mice at 12 days of age. DNA from each of 25 hepatomas, induced by N-hydroxy-2-acetylaminofluorene, vinyl carbamate, or 1'-hydroxy-2',3'-dehydroestragole, containing transforming activity in the NIH 3T3 transfection assay. Southern analysis of NIH 3T3 cells transformed by DNA from 24 of these hepatomas revealed amplified and/or rear-ranged restriction fragments homologous to a Ha-ras probe. The other tumor contained an activated Ki-ras gene. Immunoprecipitation and NaDodSO4/PAGE analysis of p21 ras proteins in NIH 3T3 transformants derived from a majority of the hepatomas suggested that the activating mutations were localized in the 61st codon of the c-Ha-ras gene. Creation of a new Xba I restriction site by an AT----TA transversion at the second position of codon 61 was detected in DNA from primary tumors and NIH 3T3 cells transformed by DNA from 6 of 7 vinyl carbamate- and 5 of 10 1'-hydroxy-2',3'-dehydroestragole-induced hepatomas. Selective oligonucleotide hybridization demonstrated a CG----AT transversion at the first position of the 61st codon in NIH 3T3 transformants derived from 7 of 7 N-hydroxy-2-acetylaminofluorene-induced hepatomas. By the same criterion, an AT----GC transition at the second position of codon 61 was the activating mutation in 1 of 7 vinyl carbamate- and 5 of 10 1'-hydroxy-2',3'-dehydroestragole-induced tumors. Thus, c-Ha-ras activation is apparently an early event in B6C3 F1 mouse hepatocarcinogenesis that results directly from reaction of ultimate chemical carcinogens with this gene in vivo.     

早期胃癌组织形态,细胞核DNA含量及免疫组织化学研究

目的:对早期胃癌病理形态、细胞核DNA含量、免疫组化及治疗等进行讨论。方法:32例早期胃癌,采用S-P法免疫组化,观察p53、p21、CEA、PCNA的表达;细胞核DNA含量以及组织病理学特征。结果:早期胃癌癌周围粘膜常伴有绒毛状异型增生、腺瘤状异型增生、囊腺样异型增生、球样异型增生、灶性异型增生的形态,并与相应腺癌一致。细胞核DNA含量,异倍体细胞在6.8％～79.5 ％之间;PCNA阳性细胞数在24.7～93.2个之间;p53、p21基因蛋白和CEA阳性表达分别为37.5％、78.1％和93.8％。早期胃癌经微波治疗后,近期经手术切除的标本中75％(6/8)仍有癌细胞存在。结论:早期胃癌与癌前期异型增生的组织学形态密切相关;异倍体值、细胞增殖指数、p53、p21和CEA表达,有助于癌前病变和早期胃癌的诊断;早期胃癌以手术切除为宜。     

Effects of endotoxin on expression of ras, p53 and bcl-2 oncoprotein in hepatocarcinogenesis induced by thioacetamide in rats

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