Function of apoptosis and expression of the proteins Bcl-2, p53 and C-myc in the development of gastric cancer

INTRODUCTION In China ,the incidence and mortality of gastric cancer rank the second among all cancers. Recent development of cancer [1-20].The aim of this study was investigat the insight of apoptosis and bcl-2, p53 and C-myc protein expression in the development of gastric cancer .     

hTERT、P53在胃癌及癌前病变中的表达及相关性研究

目的探讨端粒酶逆转录酶（hTERT）基因、p53蛋白在胃癌（GC）及癌前病变中的表达及相关性。方法采用免疫组化和原位杂交方法分别检测130例GC及癌前病变组织标本中p53和hTERT mRNA的表达。结果p53蛋白在慢性表浅性胃炎（CSG）、慢性萎缩性胃炎（CAG）、非典型增生（DYS）、GC中的表达率分别为5％、25％、50％、62．5％,其中GC与CSG、CAG比较有统计学差异：hTERT mRNA在CSG、CAG、DYS及GC中的表达率分别是0、10％、30％、78．75％,GC与CSG、CAG、DYS比较有统计学差异。p53阳性的GC组织中hTERT表达均为阳性,p53阴性的GC组织中hTERT阳性表达率为66．7％。结论hTERT是一个比p53更好的恶性肿瘤标记物;p53基因突变可导致端粒酶活化,但端粒酶激活可能不完全依赖于p53基因的调控。     

胃粘膜异型增生形态与p53,Bcl-2和c-erbB-2表达的关系

目的了解胃粘膜异型增生形态与ｐ５３，ｂｃｌ２和ｃｅｒｂＢ２表达的关系．方法胃粘膜炎症性增生４５例，异型增生４４例和胃腺癌３０例，胃粘膜活检标本进行组织形态观察，并采用ＳＰ免疫组化法，观察ｐ５３，ｂｃｌ２和ｃｅｒｂＢ２基因表达．结果异型增生病灶，小者仅有单个腺管，多者达１０余个腺管，多数３个～５个腺管．异型腺管有不同程度的扩张，为圆形或不规则形，腺上皮细胞单层或复层排列．核大小不等，染色质粗，核形态不一，为圆形、椭圆形或不规则形，可见核仁或核分裂．异型增生的腺管与周围的腺管分界清，这种现象称之为胃粘膜腺管区域性异型增生．异型增生腺管ｐ５３，ｂｃｌ２和ｃｅｒｂＢ２基因蛋白表达分别为９１％，７２７％和３８６％．结论胃粘膜腺管区域性异型增生与胃腺癌关系密切．     

原发性胃癌p53基因突变

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。     

Combined expression of gastrointestinal hormone SP and anti-apoptosis geneBcl-2 in gastric carcinoma

AIM To study the combined expression of gastrointestinal hormone substance P and anti-apoptosis gene Bcl-2 in gastric carcinoma and its significance.METHODS Substance P and Bcl-2 protein expression was examined by the S-P immunohistochemicalmethod in 33 cases of gastric carcinoma, 17 adjacent the carcinoma and 13 normal gastric mucoma.RESULTS Positive expression of SP in gastric carcinoma was higher than that of both adjacent and normalmucosa (P < 0.001). There was no statistical difference in the positive expression between adjacent andnormal mucosa (P > 0.05). The expression of bcl-2 both in gastric carcinoma and adjacent tissues werehigher than that of normal gastric mucosa (P< 0.05-0.01). But the positive expression of Bcl-2 had nostatistical significance between gastric carcinoma and adjacent tissues.CONCLUSION Both gastrointestinal hormone SP and Bcl-2 gene have synergistic expression in gastriccarcinoma, indicating that they all take part in the occurrence of gastric carcinoma. Abnormal expression ofBcl-2 gene occurred in benign gastric pathological changes, once they become carcinoma, the positiveexpression of cell is no more increased, possibly because that there is no more increase of the intensity of Bcl-2 inhibition of cell apoptosis.     

胃癌p53基因表达与转移及预后的研究

目的观察胃癌组织ｐ５３基因的超表达及其与预后的关系。方法用抗人Ｐ５３基因蛋白单克隆抗体Ｓ＿Ｐ免疫组织化学方法，观察１２８例胃癌组织ｐ５３表达状况，并对ｐ５３表达与胃癌淋巴结转移状态和术后５年生存率进行比较分析。结果胃癌组织１２８例的ｐ５３表达阳性率为４３８％（５６／１２８）；ｐ５３表达阳性和阴性组的胃癌局部和远处淋巴结转移率分别为６７９％（３８／５６）和５１４％（３７／７２），两者经统计学处理无显著性差异（Ｐ＞００５）。获得随访９８例，胃癌术后５年生存率的随访结果显示，ｐ５３阳性和阴性组分别为３８１％（１６／４２）和３０１％（１７／５６），两组间无统计学意义（Ｐ＞００５）。结论胃癌的发生与ｐ５３基因突变关系密切，并可用免疫组化检测，但Ｐ５３基因蛋白在胃癌组织中的超表达，似不能作为判断胃癌预后的参考指标，应进一步探讨     

Hp感染与胃癌和癌前病变中p53、ras、c-myc基因表达的关系

目的研究幽门螺杆菌（Hp）感染与胃癌（GC）和癌前病变中p53、ras、c-myc基因表达的关系,以探讨其致病机制。方法用美兰和W-S特殊染色方法确定Hp感染,免疫组化SP法检测p53、ras、c-myc基因的表达。结果慢性萎缩性胃炎（CAG）、肠化生（IM）、异型增生（DYS）、GC的Hp感染率均高于慢性浅表性胃炎（CSG）（P均〈0.05）;p53、ras、c-myc基因在GC、DYS中的表达均高于CAG（P均〈0.05）,p53、ras基因在IM中的表达均高于CAG（P〈0.05）;IM中Hp阳性者的p53阳性表达率高于Hp阴性者（P〈0.05）,DYS、GC中Hp阳性者p53、ras、c-myc的表达率高于Hp阴性者（P均〈0.05）。结论Hp感染可能通过调节p53、ras、c-myc基因的表达而促进GC的发生。     

Epithelial cell turnover, p53 and bcl-2 protein expression during oncogenesis of early and advanced gastric cancer in a Western population

OBJECTIVES: To investigate epithelial cell turnover alterations, and p53, bcl-2 protein expression during development of early and advanced gastric cancer in a Western population. METHODS: We investigated cell apoptosis and proliferation rates, p53 and bcl-2 protein expression in 17 early and 34 advanced gastric carcinomas and in the adjacent non-dysplastic mucosa. Cell proliferation, p53 and bcl-2 expression were detected immunohistochemically using MIB-1, anti-p53 and anti-bcl-2 monoclonal antibodies. Apoptosis was measured by TUNEL. The rate of the positive stained cells (labelling index) was count using image analysis technique. RESULTS: No difference was observed of either apoptotic (10 vs. 11) or proliferation (35 vs. 25) index between early and advanced cancers. However, the apoptotic index was significantly higher in intestinal type advanced tumors. While both apoptotic and proliferation indices were significantly higher in tumors than in the adjacent mucosa, no difference was observed of either apoptotic (2 vs. 2) or proliferation (8 vs. 13) index between the tissues adjacent to early and advanced tumors. p53 protein expression was significantly higher in advanced cancers (7 vs. 5, p=0.001) and in the non-dysplastic tissue adjacent to advanced tumors (3.5 vs. 2, p=0.001). bcl-2 labelling index was significantly higher in the mucosa adjacent to advanced carcinomas (15 vs. 5, p=0.016) but this difference did not reach significance in the tumors (20 vs. 15, p=0.37). CONCLUSIONS: Our data indicate similar cell turnover during tumorigenesis of early and advanced cancer. p53 and bcl-2 protein accumulation is more intense in gastric mucosa adjacent to advanced tumors and p53 immunoreactivity peaks in advanced carcinomas.     

p53 expression in gastric mucosa with Helicobacter pylori infection

Expression of p53 was examined immunohistochemically in the Japanese monkey model with Helicobacter pylori infection of the gastric mucosa to investigate the association between H. pylori infection and gastric carcinogenesis for a period of 4 years. In the course of these observations, from 3 years after H. pylori inoculation, nuclear staining for p53 was seen in the glandular cells of the mucosa infected with H. pylori, especially in the neck region of the glands. There was a gradual increase in the number of immunopositive cases among the infected animals. Three years after inoculation, three out of six cases, and 4 years after inoculation, four out of six cases exhibited positive staining for p53. Before inoculation, and up to 2 years after inoculation, the infected group showed no immunoreaction for p53. The non-infected group likewise displayed no immunostaining for p53 through 4 years of observation. These results suggest that p53 alterations occur in the H. pylori-infected gastric mucosa and that H. pylori infection may play an important role in gastric carcinogenesis.     

胰腺癌Bcl-2,P53蛋白表达和细胞凋亡

目的　探讨ｂｃｌ２ ,ｐ５３ 基因和细胞凋亡在胰腺癌发病机制中的作用以及它们之间相互关系．方法　应用ＡＢＣ 免疫组化技术检测５０ 例胰腺癌中Ｂｃｌ２ 和Ｐ５３ 蛋白表达,运用原位末端标记法观察肿瘤中细胞凋亡数量．结果　Ｐ５３ 蛋白表达阳性率为５４ ％ ,临床Ⅰ期阳性率（２６－７ ％ ）却显著低于Ⅱ期（６１－１ ％ ） 和Ⅲ＋ Ⅳ期（７０－６ ％ ,Ｐ＜ ０－０５） ;Ｂｃｌ２蛋白表达阳性率为６４ ％ ,临床Ⅰ期阳性率（９３－３ ％ ） ,显著高于Ⅱ期（５５－６ ％ ） 和Ⅲ＋ Ⅳ期（４７－１ ％ ,Ｐ＜ ０－０５） ;组织学Ⅲ级癌细胞中凋亡指数明显高于Ⅰ,Ⅱ级（ Ｐ＜ ０－０５） ,Ｂｃｌ２ 蛋白阴性病例中凋亡指数明显高于Ｂｃｌ２ 阳性者（ Ｐ＜ ０－０１） ．结论　Ｂｃｌ２ 是通过抑制细胞凋亡参与肿瘤的生长过程,Ｂｃｌ２和Ｐ５３ 蛋白表达之间 存在密切负相关（τ＝ － ０－１７４７ ,Ｐ＜ ０－０５） ．     

胃粘膜癌变过程中幽门螺杆菌感染与p53,c-erbB-2基因表达的研究

目的探讨胃粘膜癌变过程中幽门螺杆菌（Ｈｅｌｉｃｏｂａｃｔｅｒｐｙｌｏｒｉ,Ｈｐ）感染与ｐ５３,ｃｅｒｂＢ２基因表达的关系．方法浅表性胃炎１６例,肠上皮化生２２例,异型增生１４例,早期胃癌１８例及进展期胃癌４０例作为研究对象．用ＷａｒｔｈｉｎＳｔａｒｙ银染色法检测Ｈｐ,用免疫组化Ｓｐ法检测ｐ５３和ｃｅｒｂＢ２的基因表达产物．结果Ｈｐ,ｐ５３,ｃｅｒｂＢ２在浅表性胃炎的检出率各为５００％,００％,００％;在肠上皮化生的检出率各为５９１％,２２７％,１３６％;在异型增生的检出率各为８５７％,６４３％,２８６％;在早期胃癌的检出率各为１６７％,３３３％,１１１％;在进展期胃癌的检出率各为５０％,５２５％,５５０％;在癌旁粘膜的Ｈｐ检出率为８６７％;在癌前病变中,Ｈｐ阳性组的ｐ５３,ｃｅｒｂＢ２表达率均高于Ｈｐ阴性组．结论Ｈｐ感染参与了胃癌前病变的发生与发展;Ｈｐ感染可引起野生型ｐ５３基因失活和ｃｅｒｂＢ２基因激活,从而导致胃粘膜的癌变．     

多重受体阻滞对自发性高血压大鼠心肌细胞凋亡及其相关基因表达的影响

目的:比较多重受体阻滞与单一受体阻滞对自发性高血压大鼠(SHR)心肌细胞凋亡率(CMAR)及心肌细胞bcl-2、p53蛋白表达率的影响。方法:6周龄雄性SHR 24只,随机分4组(每组6只),在普通饲料喂养基础上,分别接受0.9%氯化钠溶液(NS)5 ml/d(对照组),卡维地洛(Car)20 mg·kg-1·d-1(Car组),缬沙组(Val)40 mg·kg-1·d-1(Val组)和Car加Val组(剂量同前)共干预8周,以2%戊巴比妥钠40 mg/kg麻醉,开胸摘下心脏,按无菌程序取左室游离壁心肌于4%多聚甲醛中固定24 h,脱水,石蜡包埋及切片,按TUNEL法及免疫组化法分别检测CMAR及bcl-2及p53蛋白表达率。结果:与对照组比较,其余各组CMAR及p53表达率明显降低,而bcl-2蛋白表达率则明显升高,上述变化在Car加Val组尤为显著,且与Car组、Val组比较差异亦有统计学意义(均P<0.05),而Car组与Val组比较,各指标均差异无统计学意义。结论:Car与Val共同产生的多重受体阻滞对SHR CMAR及其调控基因的影响优于Car或Val单独应用的效应。     

肿瘤抑制基因p53及p16与胃癌生物学行为的关系

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５１７％（３０／５８）和４８３％（２８／５８）．Ｐ５３蛋白在低分化胃癌（７００％）、进展期胃癌（５６９％）、淋巴结阳性胃癌（７４１％）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（２７３％，１４３％，３２３％）（Ｐ＜００５），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＜００５）；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（３３３％），低于淋巴结阴性胃癌中的表达率（６１３％）；相关性分析显示；ｐ５３阳性组织大多伴有Ｐ１６蛋白阳性表达（Ｐ＜００５）．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．     

胃癌nm23和P53蛋白表达及癌浸润转移的关系

目的探讨胃癌淋巴结增生转移与Ｐ５３,ｎｍ２３蛋白表达的关系．方法应用ＳＰ免疫组织化学方法,检测８７例伴有淋巴结反应性增生和（或）癌转移的胃癌进行Ｐ５３,ｎｍ２３蛋白表达．结果Ｐ５３蛋白阳性表达５２９％,淋巴结反应性增生比淋巴结伴有癌转移者阳性表达率低,差异有显著性（Ｐ＜００１）．ｎｍ２３蛋白阳性表达７７％,淋巴结反应性增生者比淋巴结伴有癌转移者阳性表达率高,差异有显著性（Ｐ＜００１）．结论Ｐ５３和ｎｍ２３蛋白阳性表达均与胃癌的淋巴结增生有关,多有相反的阳性表达,呈显著的负相关关系     

乳腺癌患者bcl-2、p53、组织蛋白酶D因子表达的临床意义

目的 观察bcl-2、p53和组织蛋白酶D（CD）在乳腺癌组织中的表达,探讨其与乳腺癌组织学分级、腋淋巴结转移及预后的关系。方法 应用免疫组化方法（SP方法）检测bcl-2、p53、CD在86例乳腺癌及33例乳腺良性病变的表达。结果 bcl-2表达与组织学分级呈负相关,其阳性表达水平可反映肿瘤属分化程度,但其与腋淋巴结转移和预后无关。p53与组织学分级呈正相关、CD与组织学分级无关;p53和CD表达与腋淋巴结转移呈正相关,与预后呈负相关。结论 bcl-2、p53和CD可作为乳腺癌组织学分级、腋淋巴结转移和预后的判断指标。     

HSP60和p53在胃癌中的表达及意义

[目的]研究热休克蛋白60(HSP60)和p53在胃癌组织中的表达,并分析与胃癌临床病理特征的关系。[方法]采用免疫组化SABC法检测48例胃癌组织和14例正常胃黏膜中HSP60和p53的蛋白表达。[结果]胃癌组织中HSP 60和p53蛋白表达阳性率分别为81.3%、68.9%;HSP60在胃癌中表达与肿瘤分化程度及增殖能力有关(P0.05);p53蛋白在胃癌中表达与肿瘤大小、部位、浸润深度、淋巴结转移和肿瘤增殖能力有关(P0.05);HSP60与p53的表达无相关性。而HSP60在正常胃黏膜中有弱阳性表达(7.1%),在正常胃黏膜中未见p53表达。[结论]HSP60和p53表达与胃癌生物学行为有关,在胃癌的发生、发展过程中发挥着重要的作用。     

Fas-mediated apoptosis with normal expression of bcl-2 and p53 in lymphocytes from aplastic anaemia

In order to investigate the involvement of apoptosis in the pathogenesis of aplastic anaemia (AA) we measured the expression of the Fas receptor (membrane protein that triggers apoptosis), Fas ligand (FasL), bcl-2 (cytoplasmatic protein that blocks apoptosis) and p53 (nuclear protein that induces apoptosis) in CD3 and CD19 lymphocytes from the peripheral blood or bone marrow of controls, patients with AA, aplastic anaemia in complete remission (AA-CR) and multiply transfused patients without aplastic anaemia. The Fas receptor was overexpressed in both T and B lymphocytes from the peripheral blood and bone marrow from patients with AA. These abnormalities were not detected in AA-CR or multiply transfused patients. CD3/FasL cells were not increased and no FasL expression was detected in B lymphocytes. Bcl-2 was highly expressed in lymphocytes from controls, AA, AA-CR and multiply transfused patients (> 99% of positive cells) whereas p53 was not detected in any group. To further characterize the functional activity of the Fas receptor we performed a Fas-induced apoptosis assay in peripheral blood lymphocytes using an anti-Fas monoclonal antibody. The crosslinking of the Fas receptor transduced an increased apoptotic signal in lymphocytes from AA patients, but not in lymphocytes from controls, AA-CR patients or multiply transfused patients. Taken together, these data suggest that a Fas-based mediated apoptosis without the apparent participation of bcl-2 or p53 is a possible mechanism of lymphocyte depletion in patients with AA. In addition, these findings suggest that Fas expression is a continuous event occurring from progenitor bone marrow cells to mature cells.     

胃癌组织芯片p53、p16及环氧合酶-2表达的研究

目的利用高通量的组织芯片技术,对胃癌组织及癌旁组织的p53、p16和环氧合酶-2（COX-2）蛋白异常表达进行分析,探讨其相关性及临床意义。方法利用组织芯片技术结合免疫组化法检测50例胃癌组织和78例癌旁组织中p53、p16及COX-2蛋白的表达。结果p53、p16和COX-2的阳性表达率在癌旁组织中分别是19％、15％及74％;在癌组织中分别是50％、54％及94％。胃癌组织中p53、p16和COX-2蛋白表达均显著高于癌旁组织,差异具有统计学意义（P〈0．05）。p53与COX-2、p16与COX-2蛋白表达均存在相关性（P〈0．05）。当p53、COX-2表达阳性,p16阴性时;或p16、COX-2表达阳性,p53阴性时;或p53、p16和COX-2同时表达阳性时,组织芯片病理类型为癌性的概率均增加,OR值分别为11．667、30．000及18．889,p53、p16和COX-2三者存在交互作用。结论联合分析p53、p16及COX-2的表达对预测胃癌的发生和发展具有重要意义。     

p53异构体和MDM2、PTEN在胃癌组织中表达的相关性

目的探讨p53的选择性剪接异构体与双微基因2(MDM2)、同源性磷酸酶张力蛋白(PTEN)在胃癌组织中的表达及其相关性。方法分别应用巢式逆转录多聚酶链反应(NT-PCR)和免疫组织化学(PV-9000两步法)方法检测p53的五种选择性剪接异构体和MDM2、PTEN在胃癌组织和癌旁胃组织中的表达,并进行统计学分析。结果在30例胃癌患者癌组织和癌旁胃组织中均未检测到三种p53异构体p53γ、Δ133p53β、Δ133p53γ的mRNA表达。与癌旁组织比较,胃癌组织中Δ133p53表达阳性率高,p53β表达阳性率低,MDM2蛋白表达阳性率高,PTEN蛋白表达阳性率低(P均<0.01)。Spearman’s相关性分析显示,Δ133p53和MDM2,p53β和PTEN在胃癌中表达均呈正相关关系(r=0.408,P=0.025;r=0.413,P=0.02);Δ133p53和PTEN,p53β和MDM2在胃癌中表达呈负相关关系(r=-0.467,P=0.009;r=-0.480,P=0.007)。结论Δ133p53、p53β通过调节p53活性,并以p53为中介,影响了PTEN-MDM2-p53网络环路中PTEN、MDM2蛋白的表达。