胃癌及癌前病变p53蛋白及Ki-67抗原检测的临床意义

应用免疫组织化学法检测胃癌和癌前病变p53蛋白、Ki-67抗原的表达.结果 正常胃黏膜无p53蛋白和Ki-67抗原表达.从萎缩性胃炎、肠上皮化生、胃黏膜异型增生至胃癌,p53蛋白和Ki-67抗原阳性表达率逐渐升高.胃癌及胃黏膜异型增生p53蛋白和Ki-67抗原阳性表达率显著高于萎缩性胃炎和肠上皮化生,差异有统计学意义（P＜0.05）;胃癌与胃黏膜异型增生、萎缩性胃炎与肠上皮化生比较p53蛋白和Ki-67抗原表达差异无统计学意义（P＞0.05）.认为p53蛋白和Ki-67抗原的检测有助于胃黏膜癌变的早期诊断.     

胆囊癌组织中Ki-67、p53的表达及意义

目的检测胆囊癌组织中Ki-67、p53蛋白的表达,并探讨其临床意义。方法采用免疫组化SP法检测42例胆囊癌组织中的Ki-67、p53。结果42例胆囊癌中Ki-67、p53的总阳性表达率分别为59．5％、38．1％。二者表达与胆囊癌的临床分期、淋巴结转移密切相关（P均〈0．05）。结论Ki-67、p53蛋白的表达与胆囊癌临床分期和淋巴结转移情况有相关性,可作为判定胆囊癌预后的独立指标。     

脑胶质瘤组织中透明质酸合成酶2表达变化及其与临床病理参数和预后的关系

目的 观察脑胶质瘤组织中透明质酸合成酶2(HAS2)的表达变化,探讨HAS2表达与脑胶质瘤临床病理参数及预后的关系。方法 收集行脑胶质瘤切除术患者的胶质瘤组织标本70例纳入病例组,肿瘤WHO分级1～2级32例、3～4级38例。另收集同期行颅内减压术的脑外伤患者的脑组织标本10例纳入对照组。分别采用免疫组化法及Western blotting法检测两组HAS2蛋白,分析HAS2表达与脑胶质瘤临床病理参数的关系。采用Logistic回归分析胶质瘤组织中HAS2表达量的影响因素。绘制Kaplan-Meier生存曲线,比较不同HAS2表达水平的脑胶质瘤患者的预后。结果 免疫组化检测结果与Western blotting检测结果均显示,病例组HAS2表达高于对照组,且3～4级脑胶质瘤组织中HAS2表达高于1～2级脑胶质瘤组织（P均<0.05）。WHO分级3～4级、突变型p53、Ki-67高表达脑胶质瘤组织中HAS2高表达率分别高于1～2级、野生型p53、Ki-67低表达的肿瘤组织（P均<0.05）。将WHO分级、Ki-67表达、p53分型、年龄、性别、肿瘤部位和O6-甲基鸟嘌呤-DN...     

p53、Ki-67、Pgp在胃癌组织中的表达及意义

采用免疫组化SP法检测52份胃癌组织石蜡切片中p53、Ki-67和PgP的表达,并分析其与胃癌临床病理特征的关系.结果 p53、Ki-67和PgP阳性表达率分别为59.6%、舳.8%、51.9%,三者阳性表达均与淋巴结转移有关(P＜0.05),Pgp阳性表达与胃癌组织浸润深度有关(P＜0.05),Pgp与p53、Ki-67联合异常表达多见于伴有淋巴结转移的进展期胃癌.认为检测p53、Ki-67和Pgp对判断胃癌淋巴结转移和选择合理的术式具有重要临床价值.     

非肌层浸润性膀胱癌组织Ki-67、p53和CK-20表达变化及其临床意义

目的 探讨非肌层浸润性膀胱癌（NMIBC）组织Ki-67、p53、细胞角蛋白20(CK-20)表达变化及其临床意义。方法 选择NMIBC患者105例,取手术切除的NMIBC组织及其配对的癌旁正常组织,采用免疫组化法检测Ki-67、p53、CK-20表达。比较NMIBC组织与癌旁正常组织Ki-67、p53、CK-20阳性表达,分析NMIBC组织Ki-67、p53、CK-20表达与患者临床病理特征的关系以及三者表达的关系。结果 NMIBC组织Ki-67、p53、CK-20阳性表达率均显著高于癌旁正常组织（χ2分别为40.485、40.752、42.091,P均<0.01）。NMIBC组织Ki-67、p53、CK-20表达与病理分级、浸润深度、术后复发有关（P均<0.05）,而与性别、年龄、肿瘤数目、肿瘤最大径无关（P均>0.05）。NMIBC组织Ki-67阳性表达与p53、CK-20阳性表达均呈正相关关系（r分别为0.642、0.669,P均<0.01）,p53阳性表达与CK-20阳性表达亦呈正相关关系（r=0.706,P<0.01）。结论 NMIBC组织Ki...     

肝癌组织中p53、Ki-67表达变化及意义

目的观察肝细胞癌（HCC）组织中p53、Ki-67蛋白表达变化,并探讨其临床意义。方法采用免疫组化PV-6000两步法检测231例HCC组织、81例癌旁组织和79例肝硬化组织中的p53、Ki-67蛋白。结果 HCC组织中p53、Ki-67蛋白阳性率为22.1%、69.7%,癌旁和肝硬化组织中均不表达,三者比较,P均〈0.01;HCC组织中p53与Ki-67蛋白表达呈正相关（r=0.147,P=0.026）。HCC组织中p53阳性者2 a累积复发率为39.2%、术后无瘤生存期为43.4个月、总体生存期为80.4个月,阴性者分别为37.8%及53.6、84.0个月,两者比较,P均〉0.05。HCC组织中Ki-67阳性者2 a累积复发率为43.5%、术后无瘤生存期为33.7个月、总体生存期为72.9个月,阴性者分别为25.7%及81.7、84.0个月,两者比较,P均〈0.05。Cox回归分析显示,Ki-67阳性是HCC肝切除术后无瘤生存的独立影响因素（R 1.583,95%CI 1.038～2.413,P〈0.05）。结论 HCC组织中p53、Ki-67蛋白表达增高,Ki-67蛋白可作为评估HCC术后早期复发的指标之一。     

C-erbB-2阴性乳腺癌组织中p53、Ki-67的表达及意义

选取57例C-erbB-2表达阴性的乳腺癌标本,采用免疫组化SP法检测其p53和Ki-67。结果显示,p53阳性表达率为40．35％,其表达与癌组织大小、临床分期、淋巴结转移及雌激素受体（ER）、孕激素受体（PR）表达情况有关。Ki-67阳性表达率为70．18％,其表达与癌组织大小及ER、PR表达情况有关。认为p53和Ki-67也可作为C—erbB-2表达阴性的乳腺癌患者的预后判断指标。     

复方水蛭素对小鼠移植瘤组织中p53、Ki-67及VEGF表达的影响

建立小鼠移植瘤模型（均接种S180瘤细胞）。取40只实验鼠随机分为四组,各10只。模型对照组给予生理盐水,阳性对照组给予环磷酰胺,实验组分别给予1.3、2.6g／kg的复方水蛭素（连续用药14d）。比较各组肿瘤生长情况,并计算抑瘤率,同时取肿瘤组织用免疫组化法检测p53、Ki-67及血管内皮生长因子（VEGF）的表达。实验组中大小剂量组抑瘤率分别为57.89％和53.68％,其肿瘤组织中p53、Ki-67阳性细胞百分数均低于肿瘤模型对照组（P〈0.05）,同时其VEGF表达降低（P〈0.05）。认为复方水蛭素对小鼠移植瘤具有显著的抑制作用,使肿瘤组织p53、Ki-67以及VEGF表达降低。     

乳腺癌肿瘤生物学因子检测对新辅助化疗方案选择的指导作用

目的探讨p53、C-erbB-2、Ki-67蛋白表达的检测对新辅助化疗方案选择的指导作用。方法采用新辅助化疗TE方案对50例Ⅱa～Ⅲb期乳腺癌患者治疗两周期,化疗前用粗针穿刺获得乳腺癌组织病理诊断,同时用免疫组化方法检测p53、C-erbB-2、Ki-67蛋白表达。结果p53蛋白表达阴性者有效率高于p53阳性者;Ki-67表达≥30％者有效率高于Ki-67表达〈30％者。结论p53、Ki-67蛋白表达水平是乳腺癌TE方案新辅助化疗疗效的重要预测因子,表达阴性者对TE方案敏感。     

组织芯片技术研究食管胃黏膜病变中Ki-67的表达及其与细胞增殖和凋亡的关系

背景：近20年来食管和食管胃连接处（EGJ）腺癌发生率呈上升趋势,Barrett食管（BE）被认为是其发生的重要危险因素。目的：应用组织芯片技术研究Ki-67在各类食管胃黏膜病变中的表达及其与细胞增殖和凋亡的关系。方法：制作140例各类食管胃黏膜组织的组织芯片,结合免疫组化方法检测Ki-67和增殖细胞核抗原（PCNA）表达。以原位末端标记技术（TUNEL法）检测BE、食管腺癌和贲门癌的细胞凋亡情况。结果：BE、食管腺癌和贲门癌组Ki-67阳性率分别为40.9%、69.6%和61.9%,均显著高于正常贲门组织（0.0%,P〈0.01）和贲门肠化生（IM）组（11.5%,P〈0.05）。BE、贲门IM、胃窦IM、食管腺癌和贲门癌组PCNA阳性率均显著高于正常贲门组织（45.5%、42.3%、39.3%、52.2%和42.9%对10.0%,P〈0.05）。BE、食管腺癌和贲门癌中Ki-67表达弱阳性和阳性者细胞凋亡指数均较表达阴性者显著降低（P〈0.05）,而PCNA仅表达阳性者凋亡指数较表达阴性者显著降低（P〈0.05）。结论：BE和EGJ处肿瘤的发生与Ki-67表达异常有关,Ki-67较PCNA更能反映EGJ处细胞凋亡状态。     

P53、malat1、ki-67和β-catenin基因mRNA检测在大肠癌分子诊断中的意义

目的:初步研究多个大肠癌相关基因p53、malat1、ki-67和β-catenin在大肠癌分子诊断中的意义.方法:收集手术切除的新鲜大肠癌组织标本47例、大肠腺瘤组织标本13例,以及分别和大肠癌、大肠腺瘤对应的正常大肠黏膜组织标本53例,用实时荧光定量RT-PCR方法检测各基因的Ct值.结果:p53、malat1在大肠癌组表达量均高于大肠腺瘤组(P=0.026,P=0.034),但是p53在正常大肠黏膜组、大肠腺瘤组和大肠癌组中表达依次增高,而malat1在大肠腺瘤组、正常大肠黏膜组和大肠癌组中表达量依次增高,大肠腺瘤组mRNA表达最低.ki-67在大肠癌组的表达量是正常黏膜组表达量的1.42倍(P=0.007).β-catenin基因在三组间的表达差异无统计学意义.各基因的表达量均和大肠癌的分期无关.经ROC曲线分析,p53的曲线下面积是0.755(P<0.05),在大肠腺瘤组和大肠癌组的最佳Cut-off值分别是2.585、3.215,malat1的曲线下面积是0.748(P<0.05),在大肠腺瘤组和大肠癌组的最佳Cut-off值分别是0.925、1.395;二元Logistic回归分析,p53和malat1进入回归模型(P<0.05).联合检测p53和malat1在大肠腺瘤组和大肠癌组的ROC曲线下面积为0.785(P=0.01),在大肠腺瘤组的和大肠癌组的最佳Cut-off值分别是0.750、0.790.p53和malat1联合检测的ROc曲线下面积均高于单独检测的ROC曲线下面积.结论:p53和malat1在大肠癌的分子诊断中有一定的应用价值.可为鉴别大肠腺瘤和大肠癌提供有效的参考;和单独检测相比,二者联和检测的准确性高于单独检测的准确性.     

大肠平坦型腺瘤、隆起型腺瘤和腺癌的p53、c-myc和ki-67表达比较

目的 检测大肠平坦型腺瘤、隆起型腺瘤和浸润性大肠癌p53、c-myc和ki-67的表达水平,探讨其基因调控机制.方法 选取2002至2006年间行结肠镜检查并经病理证实的平坦型、隆起型腺瘤和腺癌的病变标本,用免疫组织化学二步法检测p53、c-myc和ki-67的表达水平,分析三者表达水平在各组病变的差异和意义.结果 p53、c-myc和ki-67在平坦型腺瘤、隆起型腺瘤和腺癌中阳性表达率分别为3.33%(1/30)、8.06%(5/62)和69.44%(25/35);33.33%(10/30)、58.06%(36/62)和80.56%(29/36);23.33%(7/30)、19.35%(12/62)和50.00%(18/36);在正常组织中均无明显表达.结论 平坦型腺瘤、隆起型腺瘤和大肠癌三者具有不同的p53、c-myc和ki-67表达,提示三者的发生、发展可能存在不同基因调控机制,平坦型腺瘤的癌发生途径可能与隆起型腺瘤不同.     

Intrahepatic cholangiocarcinoma in Korea

We reviewed surgically treated patients with intrahepatic cholangiocarcinoma to evaluate the clinical and pathologic features of intrahepatic cholangiocarcinoma that may affect long-term survival in Korean patients with the disease. Between 1990 and 1997, 28 patients with intrahepatic cholangiocarcinoma underwent laparotomy. Resection was performed in 25 patients, and wedge resection alone in 3 patients. The liver resections consisted of right lobectomy in 5 patients, right trisegmentectomy in 1, left lobectomy in 7, extended left lobectomy in 3, hepatopancreatoduodenectomy in 2, and segmentectomy in 7. Curative resection was performed in 15 patients. Histological sections of all resected specimens were immunohistochemically stained with p53 and Ki-67 monoclonal antibodies to assess the biological behavior of the tumor cells. Cumulative survival rate and clinicopathological factors that may influence the prognosis, including biological markers (p53, Ki-67), were analyzed statistically. Patients who underwent curative resection survived significantly longer than patients who underwent noncurative resection. The median survival time of the patients who underwent curative resection was 24 months (mean, 34 ± 8 months), with 1-, 2-, and 3-year survival rates of 66.6%, 44.4%, and 35.6%, respectively. The median survival time of the patients who underwent noncurative resection was 3 months (mean, 8 ± 3 months), with 1- and 2-year survival rates of 26.7% and 13.4%, respectively. Univariate analysis showed that positive regional lymph nodes correlated significantly with poor outcome (P = 0.004) and that curative resection significantly correlated with better prognosis (P = 0.001). Age, sex, tumor size, degree of cell differentiation, gross type of tumor, and p53 and Ki-67 labeling index were not significantly correlated with outcome. Our findings support the concept that aggressive liver resection, along with regional lymph node dissection, be recommended for long-term survival. The validity of molecular biologic tumor markers (p53, Ki-67) as prognostic factors has not yet been clearly demonstrated.     

Expression of breast cancer anti‐estrogen resistance 1 in relation to vascular endothelial growth factor,p53, and prognosis in esophageal squamous cell cancer

The purpose of this study was to clarify the role of breast cancer anti‐estrogen resistance 1 (BCAR1) expression in relation to vascular endothelial growth factor (VEGF), p53, and proliferation in esophageal squamous cell cancer (ESCC). Expression of BCAR1, VEGF, p53, and the ki‐67 proliferative index were examined by tissue microarray and immunohistochemistry in 106 specimens with ESCC and matched adjacent normal tissues. Among them, 40 cases were simultaneously examined by Western blot. Both Western blot and immunohistochemistry showed that BCAR1 expression was substantially higher in ESCC than in adjacent normal tissues (P < 0.001). BCAR1 expression was significantly connected with degree of tumor differentiation, with poorly differentiated tumors showing higher BCAR1 expression (P < 0.001). BCAR1 expression was significantly and positively correlated with VEGF and p53 expression levels (r= 0.541, P < 0.001; r= 0.374; P < 0.001) but not proliferative index (r= 0.44; P= 0.066). Additionally, a significant relationship was also observed between VEGF and p53 (r= 0.321; P= 0.001). Kaplan–Meier survival analysis revealed that patients with high BCAR1 expression had significantly shorter survival times than those with low BCAR1 expression levels (median survival 40 months vs. 27 months, P= 0.09). Multivariate analysis also revealed that levels of BCAR1 expression (hazard ratio 2.250, P= 0.015) was a significant and independent prognostic indicator. High expression of BCAR1 is associated with elevated VEGF and p53 expression levels, as well as poor prognosis in ESCC. Therefore, BCAR1 may be a potential candidate for predicting prognosis and a new therapy target for ESCC.     

p53、Ki-67基因表达与膀胱癌生物学特性的研究

目的 探讨p53、Ki-67基因表达与膀胱移行细胞癌(BTCC)病理分级、临床分期及复发的关系.方法 收集149例BTCC患者的临床资料及p53、Ki-67表达阳性率.结果 Ⅰ～Ⅱ级BTCC p53、Ki-67阳性率分别为58.8%、64.7%,低于Ⅲ级BTCC p53、Ki-67的阳性率(80.014%、90.0%)(P<0.05,P<0.01);浸润型BTCC的p53、Ki-67基因表达阳性率(80.0%、92.3%)均高于浅表型BTCC阳性率(50.0%、52.4%)(P<0.01).在高分级、高分期BTCC组p53、Ki-67联合表达率(73.3%、73.8%)高于低分级、低分期BTCC组(45.4%、32.1%)(P<0.05).p53、Ki-63共同表达阳性者复发率为37.2%,单一阳性或阴性表达者复发率16.1%,两组复发率有显著性差异(P<0.05).结论 p53、Ki-67基因表达与BTCC的临床分期、病理分级相关,提示p53、Ki-67蛋白过度表达的BTCC高度恶性和预后不良;p53、ki-67基因共同表达阳性组复发率较单一阳性或阴性组复发率高,提示p53和ki-67基因联合表达与BTCC复发可能有相关性.     

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。     

Immunohistochemical expression of cell-cycle proteins in gastric precancerous lesions

Background: The early indicator for the subject predisposed to gastric cancer is abnormal proliferation of gastric epithelial cells, such as atrophic gastritis (AG), intestinal metaplasia (IM), and dysplasia, which have been considered as precancerous lesions of gastric cancer. To determine whether p53 protein, cyclins D1, and D3, and p27^kip1 play a role in the carcinogenesis pathway of gastric cancer, we performed an immunohistochemical study of their expression in gastric precancerous lesions. Methods: A total of 1 45 endoscopic gastric biopsy specimens of AG, IM, and gastric dysplasia were studied. These molecular markers were localized by immunohistochemistry. Results: P53 was expressed in 15% of cases with gastric dysplasia and not in the pre‐dysplastic stages of the gastric mucosa. All cases were concerning high‐grade dysplasia. Cyclin D1 protein was almost undetectable in the precancerous lesions of gastric cancer. Cyclin D3 protein overexpression was seen in 10% of biopsies with IM, and 50% of biopsies with gastric dysplasia. High expression of p27^kip1 protein was demonstrated in all cases of chronic gastritis. As atrophy, IM, and dysplasia develop, expression of p27^kip1 protein is suppressed. In total, 15% of dysplastic cases showed no expression of p27^kip1 protein. Conclusions: (i) P53 mutation must be a late event during the development of gastric cancer. (ii) Cyclin D1 protein overexpression may not play a role in the progression from normal to neoplastic gastric mucosa, while overexpression of cyclin D3 is an earlier event during gastric carcinogenesis, and its role must be further evaluated. (iii) Reduced expression of p27^kip1 is a rather early event in gastric tumorigenesis, before dysplastic changes occur.     

P53 and Ki-67 overexpression in gastroesophageal reflux disease – Barrett's esophagus and adenocarcinoma sequence

Gastroesophageal reflux disease (GERD) is a major risk factor for the development of esophageal adenocarcinoma (ACE). Many molecular alterations occur in esophageal carcinogenesis, yet the exact mechanism of ACE development remains unknown. This study aims to determine p53 protein and Ki-67 expression in esophageal mucosa of patients with GERD and study the correlation between these markers and the progression from normal squamous epithelium to esophagitis, columnar epithelium with or without intestinal metaplasia and ACE. We analyzed p53 protein and Ki-67 expression in biopsies of 200 patients with GERD and 35 patients with ACE. Those biopsies were classified into five groups: (i) G1 normal squamous epithelium (58); (ii) G2 esophagitis (80); (iii) G3 columnar epitheliums without intestinal metaplasia (30); (iv) G4, columnar epitheliums with intestinal metaplasia (32); and (v) G5 ACEs (35). p53 protein overexpression was found in 7% (4) of G1, 37.5% (30) of G2, 30% (9) of G3, 62.5% (20) of G4, and 71.4% (25) of G5 (p < 0.001). Ki-67 index increased according to the severity of histopathological diagnoses. Ki67 index was 21.3 ± 19.5% in G1, 38.8 ± 24.9% in G2, 37.7 ± 26.3% in G3, 52.8 ± 24.6% in G4, and 57.1 ± 25.1% in G5 ( P < 0.001). Linear correlation between p53/Ki67 expression and the multistep progression from squamous epithelium to ACE was observed ( P < 0.001 and P < 0.05). Our results indicate that overexpression of p53 and increased Ki-67 could be associated with the development and progression to ACE in patients with GERD.