食管癌中HPV16、18感染与多癌基因产物表达的研究

目的：探讨高危型ＨＰＶ１６、１８感染、多个癌基因激活在食管癌发生机制中的作用。方法：采用免疫组化技术,检测本地区人口食管鳞癌、食管鳞状上皮不典型性增生和食管粘膜慢性炎症组织中ＨＰＶ１６、１８Ｅ６蛋白,Ｐ５３,ｐ２１ｒａｓ,ｃ－ｍｙｃ和ｃ－ｅｒｂＢ－２癌基因蛋白的表达情况,结果：癌组中Ｅ６、ｐ５３、ｐ２１ｒａｓ、ｃ－ｍｙｃ、ｃ－ｅｒｂＢ－２和不典型性增生组中Ｅ６、ｐ５３、ｐ２１ｒａｓ癌基因蛋白的阳性     

人膀胱移行细胞癌HPV16／18型感染与c—erbB2,H—ras,c—my …

目的：研究人膀胱移行细胞癌（ＴＣＣ）中ＨＰＶ－１６／１８型感染与ｃ－ｅｒｂＢ２、Ｈ－ｒａｓ、ｃ－ｍｙｃ蛋白产物表达的相互关系。方法：应用免疫组织化学法检测经聚合酶链反应证实的３４例ＨＰＶ－１６／１８感染阳性、２０例ＨＰＶ－１６／１８感染阴性的ＴＣＣ组织和７例正常膀胱组织中ｃ－ｅｒｂＢ２、Ｈ－ｒａｓ、ｃ－ｍｙｃ蛋白产物的表达,并经统计学处理。结果：ＨＰＶ－１６／１８感染阳性组ｃ－ｅｒｂＢ２、Ｈ－ｒａ     

EBV感染、p53、Bcl-2、C-myc基因表达与肺癌关系研究

目的 研究肺癌组织中ＥＢＶ感染率以及ｐ５３、Ｂｃｌ－２和Ｃ－ｍｙｃ基因的表达,分析ＥＢＶ和ｐ５３、Ｂｃｌ－２、Ｃ－ｍｙｃ基因表达的关系。方法 检测４８例手术切除肺癌标本,１８例癌旁支气管粘膜组织,２例肺转移平滑肌肉瘤,１例结核瘤,１４例正常肺组织中ＥＢＶ ＤＮＡ及ｐ５３、Ｂｃｌ－２、Ｃ－ｍｙｃ基因表达。用ＰＣＲ法检测新鲜组织的ＥＢＶ ＤＮＡ,间接原位ＰＣＲ法观察ＥＢＶ阳性信号在细胞中的反应部位,免     

胃癌p53和c—erbB—2基因产物的表达及其与病理临床的相关性研究

用免疫组化ＡＢＣ法对９２例胃癌的石蜡包埋标本进行了ｐ５３和ｃ－ｅｒｂＢ－２基因蛋白表达的研究。结果显示：９２例胃癌ｐ５３、ｃ－ｅｒｂＢ－２蛋白表达的阳性率分别为３８．０％和１４．１％；ｐ５３表达主要位于细胞核内，而ｃ－ｅｒｂＢ－２表达主要位于细胞膜，ｐ５３及ｃ－ｅｒｂＢ－２表达与胃癌患者的性别、年龄、原发灶部位、淋巴结及远处转移、肿瘤分期以及病理类型、分级均无明显关系。ｐ５３阳性表达的胃癌患者生存期明显低于ｐ５３阴性者，而ｃ－ｅｒｂＢ－２表达与胃癌患者的生存期无关；ｐ５３与ｃ－ｅｒｂＢ－２二者阳性表达之间亦无关。５例ｐ５３及ｃ－ｅｒｂＢ－２表达的阳性者预后极差，提示，癌基因的异常表达对预后的估计具有辅助意义。     

肺癌诊断的基因标志

本文系统地介绍了与肺癌生成密切相关的ｒａｓ，ｍｙｃ，ｅｒｂＢ基因家族等原癌基因的多种激活方式，以及抑癌基因ｐ５３，Ｒｂ和ｃｈｒ３ｐ上特殊基因的结构缺失和异常。同时，还从分子水平深入探讨了肺癌生成中ｐ５３，ｒａｓ基因频发基因点突变的类型和性质，以及它们做为基因标志在肺癌诊断中的意义。     

癌基因rasp^21,c—myc和p^53在乳腺增生与乳腺癌中的表达相…

应用Ｓ－Ｐ微波免疫组化法检测乳腺单纯性增生，不典型增生各２０例，乳腺癌４０例中ｒａｓｐ２１、ｃ－ｍｙｃ、Ｐ５３基因产物的表达，结果：ｒａｓｐ２１在不典型增生和癌中阳性表达率均较高，在癌中与ｃ－ｍｙｃ、Ｐ５３基因的联合表达率亦高，表达Ｐ２１基因在细胞早期癌变过程中具有引发作用，为癌的启动基因，ｒａｓ基因的激活 癌需有其他癌基因的协同，和ｃ－ｍｙｃ基因互补，ｃ－ｍｙｃ和Ｐ５３在乳腺癌及伴有转移的癌中表     

男性乳腺癌癌基因与抗癌基因表达的初步研究

本文观察了１２例男性乳腺癌的癌基因与抗癌基因产物的表达。１２例中ｃ－ｍｙｃ阳性６例，ＥＧＦＲ阳性６例，ｃ－ｅｒｂＢ－２阳性４例，Ｎ－ｒａｓ阳性２例，Ｒｂ阳性５例，ｐ５３阳性３例。上述癌基因与抗癌基因产物中，全部阳性的１例，全部阴性的４例。此外还作了ＣａｔｈｅｐｓｉｎＤ、ＥＲ、ＰＲ、ＡＲ、ＰＣＮＡ与ＡｇＮＯＲ检测，ｃ－ｅｒｂＢ－２或ｐ５３阳性的病例，ＣａｔｈｅｐｓｉｎＤ均阳性。     

肺癌患者癌组织和痰液细胞中p53和K-ras基因突变的研究

目的 检测肺癌组织和肺癌患者痰液脱落细胞中ｐ５３、Ｋ－ｒａｓ基因突变情况,比较联合检测ｐ５３、Ｋ－ｒａｓ和单一检测ｐ５３或Ｋ－ｒａｓ基因在肺癌诊断中的价值。方法 应用ＰＣＲ－ＳＳＣＰ－银染法检测了５９例肺癌组织、癌旁肺组织、１４全肺部良性病变肺组织及患者痰液脱落细胞中ｐ５３基因第５～８外显子、Ｋ－ｒａｓ基因第１外显子突变。结果 肺癌组织中ｐ５３基因突变率为３７．３％（２２／５９）,痰液脱落细胞为３     

胃癌多基因表达的同步检测

应用ＬＳＡＢ和ＡＢＣ法对７５例胃癌及癌旁组织进行了ｐ５３、ｃ－ｅｒｂＢ－２、ＥＧＦＲ和ｒａｓ表达的研究。结果显示：（１）７５例胃癌ｐ５３、ｃ－ｅｒｂＢ－２、ＥＧＦＲ和ｒａｓ表达阳性率分别为４１．３％、１８．７％、６１．３％和４６．７％。ｐ５３在肠型胃癌中阳性率为５２．６％，高于弥漫型胃癌的２９．７％（Ｐ＜０．０５）；在早期胃癌中阳性率较高（６０．０％），癌旁重度异型增生亦有阳性表达（２／４）。ｃ－ｅｒｂＢ－２阳性表达只限于癌灶，而癌旁组织均为阴性。ｃ－ｅｒｂＢ－２在高分化胃癌中阳性率较高（Ｐ＜０．０５）。ＥＧＦＲ表达与胃癌的大体类型、生长方式、分化程度、淋巴结受累和远处转移呈正相关（Ｐ＜０．０５）。（２）ｃ－ｅｒｂＢ－２和ＥＧＦＲ在胃癌中的表达互相独立。（３）ｒａｓ表达与ＥＧＦＲ表达呈正相关，而与ｃ－ｅｒｂＢ－２呈负相关。（４）ｐ５３表达与其它基因表达无明显关系。上述结果表明，胃癌发生发展过程中伴有多种癌基因的变化，其出现时间不同，意义也不一样。     

ras和erbB2基因在肺癌中的表达及意义

目的：了解ｒａｓ和ｅｒｂＢ２基因在肺癌中表达的临床意义。方法：采用免疫组化技术研究了ｒａｓｐ２１和ｅｒｂＢ２ｐ１８５在４０例肺癌（腺癌１９例，鳞癌１８例，腺鳞癌１例，小细胞肺癌２例）中的表达，并将肺癌按临床病理特征分组进行对比分析。结果：①在非小细胞肺癌（ＮＳＣＬＣ）中，ｐ２１的阳性率为６１％，ｐ１８５的阳性率为４２％，２例小细胞肺癌（ＳＣＬＣ）均无ｐ２１和ｐ１８５的表达。②ｐ１８５在肺腺癌中的表达明显高于鳞癌（Ｐ＜０．０５）。③在ＮＳＣＬＣ中，ｐ２１在Ⅰ～Ⅱ期中的阳性率为２５％，Ⅲａ～Ⅲｂ期中的阳性率为７７％，两者有非常显著的差异（Ｐ＜０．０１）。④ｐ２１和ｐ１８５阳性的ＳＮＣＬＣ患者，其淋巴结转移发生早，速度快（Ｐ＜０．０５）。⑤ｐ１８５在ＳＮＣＬＣ中的表达存在性别差异，女性明显高于男性（Ｐ＜０．０１）。结论：ｒａｓ和ｅｒｂＢ在ＮＳＣＬＣ中的表达具有重要的临床意义。     

非小细胞肺癌分子病理学的研究

目的　探讨上海地区人肺癌发生及发展过程中的分子病理学模式。方法　采用DNAslotblot、PCR、PCR SSCP等方法 ,先后检测 2 0 0例NSCLC组织DNA标本中C erbB2、C myc、EGFR癌基因扩增 ,抑癌基因中p5 3基因外显子 5～ 8点突变、p15基因的纯合性丢失 ,以及某些与肺癌相关的染色体 3p及 17p13 .3位点的杂合性丢失 (LOH)。结果　多种癌基因共扩增率与肺癌TNM分期呈正相关 (P <0 .0 0 1)。p5 3基因外显子 5～8点突变率为 49.2 % ( 3 1/63 ) ,其中以外显子 8为主。肺癌组织中p15基因出现高频率纯合性丢失 ,并与肺癌TNM分期密切相关 (P <0 .0 0 5 )。 17p13 .3杂合性丢失率为 40 % ( 8/2 0 ) ,并与p5 3基因的点突变相关联。 3p14及 3p2 5位点处的杂合性总丢失率可高达 66.7% ( 10 8/162 ) ,其中肺腺癌中 3p14及 3p2 5二位点的共丢失率明显高于鳞癌 (P <0 .0 5 )。结论　根据上述结果 ,提出上海市区居民的非小细胞肺癌分子病理学的初步模式 :癌前期病变时已可出现 3p丢失 ,原位癌时以p5 3基因突变及 17p13 .3丢失为主 ,浸润癌及向转移性癌过渡时有多种癌基因C myc、C erbB2及EGFR癌基因共扩增 ,并出现p15基因纯合性丢失。     

端粒酶与p53、p16基因在非小细胞肺癌中的表达及其意义

目的研究端粒酶与p53、p16基因在非小细胞肺癌(NSCLC)中的表达及其意义.方法应用SP法免疫组化技术和TRAP法分别检测40例肺癌组织中端粒酶和p53、p16基因的表达.结果端粒酶及p53、p16基因在NSCLC中的阳性率分别为92.5%、37.5%和66.7%;p53基因阳性表达率与NSCLC及病理分期显著相关(P＜0.05).p16基因阳性表达率与性别、年龄、病理分期、吸烟史、组织学类型和分化程度无显著相关(P＞0.05).端粒酶活性与年龄呈负相关,并与p53基因表达情况和病理分期有关(P＜0.05).结论端粒酶和p53基因的表达与NSCLC的病期进展有关.     

周围型肺癌CT征象与p53蛋白及增殖细胞核抗原表达间关系的研究

目的研究周围型肺癌ＣＴ征象与ｐ５３蛋白及增殖细胞核抗原（ＰＣＮＡ）表达间的关系。方法运用ＳＰ免疫组化法,检测３２例经病理证实的周围型肺癌组织中ｐ５３蛋白及ＰＣＮＡ的表达,并回顾性分析其与术前ＣＴ征象间的关系。结果ｐ５３蛋白、ＰＣＮＡ表达与瘤体大小、深分叶征、空洞、胸膜凹陷征、纵隔淋巴结转移有关,与毛刺征无关。结论周围型肺癌ＣＴ征象中,瘤体直径＞３ｃｍ,或出现深分叶征、空洞、胸膜凹陷征、纵隔淋巴结转移者,具有相对更高的恶性程度,肿瘤细胞的增殖更为活跃。     

Prognostic significance of K-ras,p53, bcl-2, PCNA,CD34 in radically resected non-small cell lung cancers

The aim of this study was to investigate the prognostic significance of a panel of biological parameters in patients with radically resected non-small cell lung cancers (NSCLC). 269 cases with pathological stage I-IIIA NSCLC were retrospectively analysed. Immunohistochemistry was performed to detect protein expression of p53, bcl-2, proliferating cell nuclear antigen (PCNA) and CD34. Polymerase chain reaction (PCR)/direct nucleotide sequencing method was used to detect mutations in K-ras (codons 12, 13, 61, exons 1-2). The Kaplan-Meier estimates of survival were calculated for clinical and biological variables using the Cox model for multivariate analysis. Histological subtype and the pathologic tumour extension (pT) were the most powerful clinical-pathological prognostic factors for survival (P=0.030 and P=0.031, respectively), whereas among the biological parameters, p53 overexpression (P=0.032) and K-ras mutation (P=0.078) had a negative prognostic role, as demonstrated by multivariate analysis. Conversely, bcl-2, PCNA and CD34 expression were not correlated with survival. Statistically significant associations between p53 expression and the squamous cell carcinoma (SCC) subtype, bcl-2 expression and SCC subtype, K-ras mutation and p53 negative expression, p53 and bcl-2, bcl-2 and PCNA overexpression were observed. In conclusion, some biological characteristics such as the K-ras and p53 status may provide useful prognostic information in resected NSCLC patients, in addition to the classical clinico-pathological parameters. However, further studies are needed to clarify the value of adopting biological prognostic factor into clinical practice.     

The correlation between the expression of multidruc resistance related gene and cell apoptosis and clinical significance in non-small cell lung cancer

Objective: To explore the correlation and clinical significance between expression of MDR (multidrug resistance) related gene MRP, MDR₁, C-erbB-2 and cell apoptosis in non-small cell lung cancer (NSCLC). Methods: RT-PCR, Immunohistochemistry were used to examine the expression of mRNA and protein in the MDR and apoptosis related gene. Apoptosis cells were assayed by Terminal deoxynucleotidyl transferase (TdT)- mediated biotin dUTP nick end-labeling (TUNEL). Results: The positive rates of MRP, MDR₁, C-erbB-2, bcl-2, C-myc mRNA in 63 cases NSCLC were 81.0% (51/63), 38.1%(24/63), 47.6%(30/63), 65.1% (41/63), 76.2% (48/63) respectively. Their levels were higher than those of corresponding proteins (74.6%, 34.9%, 46.0%, 61.9%, 71.4%, respectively). The significant association was found between the mRNA level and the protein expression (r=+0.764,P<0.02). The C-myc expression in 2 cases adjacent and benign lung tissue were light positive, and another 3 cases were negative. The positive correlation were demonstrated between C-myc and C-erbB-2 (r=+0.547, p=0.001) as well as bcl-2 and C-erbB-2 (r=+0.486, p=0.023) in NSCLC. There is no any correlation among bcl-2, C-myc and MRP or MDRI. There exists inverse correlation between apoptotic index and bcl-2 (r=−0.587, p=0.017), and no any correlation among apoptotic index and MRP or MDRI or C-erbB-2 or C-myc. The average apoptotic index were higher in the effective chemotherapy group (27.2±2.1, 30.5±1.8) than that in the non-effective chemotherapy group (9.4±1.3, 12.6±2.4) with adenocarcinoma and squamous cell carcinoma (p=0.01, p=0.004). The positive rates of bcl-2, MRP, C-erbB-2 expression in the effective chemotherapy group (31.8%, 40.9%, 22.7%, respectively) were lower than those in the non-effective chemotherapy group (77.4%, 90.3%, 67.7%, respectively) (p=0.036, p=0.012, p=0.01), but MDR₁ and C-myc expression have no any significant difference (p=0.067, p=0.282). The median survival time in the patients with coexpression of more than three MDR and/or apoptosis related genes are shorter (8.6 months) than that in those patients with coexpression of less than three MDR and/or apoptosis related genes (15.5 months)(p=0.01). Conclusion: The multidrug resistance in NSCLC is not only related to many drug resistance genes, but also involved in cell apoptosis and apoptosis related gene expression. The coexpression of MDR and apoptosis related gene is related to the survival time. This work was supported by the Grant from Beijing Natural Science Foundation(No.7992005), and a Grant from Postdoctoral Foundation of National Committee of Education of China.     

子宫内膜样腺癌组织中P53、P63和C-erbB2的表达及其意义

背景与目的:p53与C-erbB2是已被证实与子宫内膜样腺癌(endometrioidadenocarcinoma,EC)密切相关的抑癌基因和癌基因,而它们之间的相关性报道较少。p63在结构上与p53高度同源,被认为是p53突变时的一种抑癌基因,它的抑癌特性尚未确定,它在EC中的表达少见文献报道。本研究旨在探讨p53、p63和C-erbB-2基因在EC发生发展中的作用及其与EC临床病理特征的关系。方法:采用免疫组化SP法检测38例EC和23例子宫内膜增生过长(endometrialhyperplasia,EH)及10例正常增生期子宫内膜(benignproliferativeendometrium,BPE)中P53、P63和C-erbB2蛋白的表达情况。结果:(1)P53蛋白在EC组中的阳性率为31.6%,明显高于EH组和BPE组(P<0.05)。P53的表达与EC的手术病理分期和肌层浸润深度有关(P<0.005),而与组织学分级无关(P>0.05)。(2)P63蛋白在EC组中的阳性率为81.6%,与EH组、BPE组比较,差异均有统计学意义(P<0.005)。P63的表达与EC的组织学分级、手术病理分期和肌层浸润深度均无关(P>0.05)。(3)C-erbB2蛋白在EC组中的阳性率为23.2%,与EH组、BPE组比较,差异均无统计学意义(P>0.05)。C-erbB2的表达与EC的手术病理分期和肌层浸润深度有关(P<0.001,P<0.005),而与组织学分级无关(P>0.05)。(4)P53和P63在EC中的表达呈正相关(r=0.443,P<0     

多基因表达对IIIA期非小细胞肺癌术后转移及生存期的影响

目的前瞻性探讨抑癌基因p53,癌基因K-ras、HER2, 血管内皮生长因子(VEGF),表皮生长因子受体(EGFR),粘连因子CD44,金属蛋白酶MMP9等蛋白表达对ⅢA期非小细胞肺癌(NSCLC)术后复发转移和生存期的影响.方法采用免疫组织化学方法检测32例ⅢA期非小细胞肺癌手术标本中上述蛋白表达.结果 p53、K-ras、VEGF、EGFR、CD44、MMP9和HER2蛋白表达率分别为62.5%(20/32)、34.3%(11/32)、25.0%(8/32)、46.9%(15/32)、78.1% (25/32)、50%(16/32)和43.8%(14/32).K-ras,EGFR蛋白表达与疾病进展时间(time to progression,TTP)有关(P=0.030, P=0.008).多变量Cox回归分析,K-ras,HER2蛋白表达是影响生存期的独立影响因素.Kaplan-M 生存分析显示K-ras,HER2 阳性组生存期明显短于阴性组(P=0.042,P=0.039).结论对于ⅢA期NSCLC,上述7种因子阳性表达率很高.K-ras,EGFR蛋白表达阳性患者术后更易发生远处转移, K-ras,HER2表达是影响生存期的独立影响因素.     

THE CORRELATION BETWEEN THE EXPRESSION OF MULTIDRUG RESISTANCE RELATED GENE AND CELL APOPTOSIS AND CLINICAL SIGNIFICANCE IN NON-SMALL CELL LUNG CANCER

The mechanisms about multidrug resistance (MDR) which are studied widely in present include overexpression of multidrug resistance protein such as MDR1/P-gp, MRP (multidrug resistance related protein), LRP (lung resistance protein), increased detoxification of Glutathione / Glutathione - S - transferase II.[1-3] Recent studies have showed that inhibition of cell apoptosis and overexpression of apoptosis related gene is another reason for the MDR.[4] But there are few reports about the …     

Analysis of K-ras, p53, bcl-2 and Rb expression in non-small cell lung cancer cell lines

Six non-small cell lung cancer (NSCLC) cell lines (A-549, Ca-Lu-6, SK-Lu-1, Ca-Lu-1, SK-Mes-1 and LX-1) were studied to assess the presence of multiple concomitant alterations of different oncogenes (K-ras, bcl-2) and tumor suppressor genes (p53, Rb) in NSCLC. K-ras (exon 1) and p53 (exons 5-8) gene mutations were determined via a PCR-based-DGGE (Denaturing Gradient Gel Electro-phoresis) and by sequencing approach. Different mutations were found in the Ist exon of K-ras gene in 5 of 6 cell lines examined. Five of six cell lines contained K-ras mutations at codon 12 (A-549, SK-Lu-1, LX-1) or codon 13 (SK-Mes-1, Ca-Lu-1). In addition, 5 of 6 cell lines showed p53 mutations of exon 8 (SK-Mes-1, Ca-Lu-1 cod. 280; LX-1 cod. 273) or exon 6 (Ca-Lu-6 cod. 196; SK-Lu-1 cod. 193). In 4 of these cell lines, p53 protein nuclear expression was also confirmed with DO-7 mAb immunocytochemistry. Expression of cytoplasmic bcl-2 protein, by anti-bcl-2 mAb flow cytometric analysis, was found in A-549, Ca-Lu-1, SK-Lu-1, SK-Mes-1 cell lines. In contrast, RT-PCR analysis of Rb gene could not identify any change in the cell lines examined. In conclusion, most NSCLC cell lines tested displayed concomitant multiple oncogene/tumor suppressor gene alterations.