结直肠癌中NKD1和β-catenin的表达及意义

目的观察NKD1和β-连环素(β-catenin)在结直肠正常黏膜组织、结直肠腺瘤及结直肠癌中的表达,探讨两者在结直肠肿瘤发生、发展中的作用。方法应用免疫组化SP法检测NKD1和β-catenin在正常黏膜组织、结直肠腺瘤、结直肠癌组织中的表达。结果 NKD1在正常组织中的阳性率明显高于结直肠腺瘤、结直肠癌,其表达与肿瘤的分化程度、Dukes分期及淋巴结转移相关,差异有统计学意义(P<0.05)。β-catenin在癌组织中的表达异常率明显高于癌旁正常组织,其异位表达率与肿瘤的分化程度、Dukes分期、淋巴结转移有关,差异有统计学意义(P<0.05)。NKD1的表达和β-catenin异位表达呈负相关(rs=-0.691,P<0.01)。结论 NKD1的表达下调与β-catenin的异位表达,可能与结直肠肿瘤的发生、发展有关。     

环氧合酶-2在预测结直肠癌肝转移中的价值

目的研究环氧合酶-2（COX-2）在结直肠组织中的表达，探讨其与结直肠癌肝转移的关系。方法免疫组化学检测无肝转移、术时合并肝转移和术后发生肝转移的各30例结直肠组织以及30例癌旁正常粘膜组织（对照组）中的COX-2表达水平，分析其与性别年龄、Dukes分期等临床特征之间的联系，比较无肝转移、术时肝转移、术后肝转移的结直肠癌组织和癌旁正常粘膜组织中的COX-2表达水平。结果COX-2表达情况：在结直肠组织中的表达显著高于癌旁正常粘膜组织（P〈0.05）；在术时肝转移和术后肝转移组中的表达均显著高于无肝转移组（P〈0.05）；在结直肠癌组织中的表达与性别、年龄、分化登记、病理类型、术前癌胚抗原（CEA）和糖链抗原19-9（CA19-9）无关，与Dukes分期、淋巴结转移有关。结论COX-2过度表达与结直肠癌发生、发展、肝转移相关，对结直肠肝转移早期诊断有一定的确应用价值。图4表2参8。     

结直肠腺癌中TKT的表达及临床意义

目的探讨结直肠腺癌中转酮醇酶(transketolase, TKT)的表达及与临床病理特征的关系。方法通过Oncomine和GEO数据库分析TKT基因在结直肠腺癌和癌旁组织中的表达。收集配对结直肠腺癌及癌旁正常肠黏膜组织,分别采用qRT-PCR、Western blot法检测TKT mRNA和蛋白表达,运用免疫组化法检测TKT蛋白表达,并分析其与临床病理特征的相关性。结果 qRT-PCR结果显示:与癌旁组织相比,27例结直肠腺癌组织中TKT mRNA表达显著升高(P<0.001);Western blot结果显示:8例结直肠腺癌组织中TKT蛋白表达高于癌旁组织,在常见结直肠癌细胞株中TKT蛋白表达高于FHC细胞株;两种检测与数据库样本分析一致。免疫组化检测结果显示:92例结直肠腺癌组织中TKT高表达率显著高于癌旁组织(P<0.001),TKT高表达与肿瘤直径(P=0.003)、脉管侵犯(P=0.021)、淋巴结转移(P=0.005)及T分期(P=0.007)相关。结论 TKT在结直肠腺癌中呈高表达,与肿瘤侵袭转移临床病理特征密切相关,其可能参与结直肠腺癌的发生、发展,TKT有望成为结直肠癌预后标志物或潜在的治疗靶点。     

结直肠癌中Survivin、Caspase 9和Ki-67表达的相关性及临床意义

目的探讨Survivin、Caspase 9和Ki-67三种肿瘤相关因子在结直肠癌中表达的相关性。方法采用免疫组化Eli Vision法检测50例结直肠癌、30例结直肠腺瘤和30例癌旁正常组织中Survivin、Caspase 9和Ki-67的表达,并分析三者与结直肠癌临床病理特征的关系。结果 Survivin和Ki-67在结直肠癌组织中的阳性率高于结直肠腺瘤和癌旁正常组织;而Caspase 9阳性率低于结直肠腺瘤和癌旁正常组织。Survivin与Caspase 9的表达呈负相关,与Ki-67表达呈正相关;Caspase 9与Ki-67的表达呈负相关。Survivin、Ki-67、Caspase 9的表达与结直肠癌分化程度、淋巴结转移及Dukes分期相关(P0.05)。结论Survivin可能通过抑制Caspase 9的激活,使细胞不能进行有效凋亡并异常增殖,从而促进结直肠癌的发生、发展。联合检测有助于了解结直肠肿瘤的生物学行为并对患者早期诊断、预后判断及干预治疗提供分子生物学支持。     

NPM1在结直肠癌的表达及临床价值

目的 观察核仁磷酸蛋白（NPM1）在结直肠中的表达及其意义.方法 用免疫组化法对50例结直肠癌患者癌组织和癌旁组织中的NPM1表达进行分析.结果 NPM1在结直肠癌组织中的表达显著高于癌旁组织（P ＜0.001）;NPM1表达上调与肿瘤直径、TNM分期、Dukes分期、淋巴结转移及远处转移显著相关,而与患者性别、年龄及结直肠癌分化水平无关（P＞0.05）.结论 NPM1的表达与结直肠癌的发生发展有关.     

结直肠腺癌中MMP-19的表达及意义

目的观察基质金属蛋白酶-19(matrix metalloproteinase-19,MMP-19)在结直肠腺癌组织中的表达及意义。方法采用免疫组化法检测71例结直肠腺癌、32例结直肠腺瘤和34例癌旁正常结肠黏膜组织中MMP-19的表达,并复习相关文献。结果 MMP-19在结直肠腺癌、腺瘤和正常结肠组织中的阳性率分别为40. 84%、40. 63%、8. 82%,三组间差异有统计学意义(P=0. 003)。结直肠癌中MMP-19的表达与肿瘤最大径、部位、分化程度、浸润深度、淋巴结转移、有无癌栓、有无癌结节均无明显相关性(P 0. 05)。结论MMP-19在结直肠腺癌中高表达,提示其在结直肠腺癌发生过程中可能具有重要促进作用。     

Th22细胞及相关因子在结直肠癌的表达

目的通过检测Th22细胞及相关因子在结直肠癌的表达,探讨Th22细胞在结直肠癌发病机理中的作用。方法采用流式细胞技术检测50例结直肠癌患者肿瘤组织以及癌旁组织中Th22细胞比例的表达差异,同时采用qRT-PCR方法检测IL-22、AHR、CCL27、CCL22、CCL20在结直肠肿瘤微环境中的mRNA相对表达量。结果结直肠肿瘤组织中的Th22细胞比例明显高于癌旁组织(P0.05),Dukes A+B期患者的Th22细胞比例显著低于Dukes C+D期(P0.05);qRT-PCR结果显示IL-22、AHR、CCL22C、CL20在肿瘤组织的mRNA表达水平明显高于癌旁组织(P0.05),而CCL27在肿瘤组织的表达水平也较癌旁组织增高,但无统计学差异(P0.05)。结论 Th22细胞在结直肠恶性肿瘤中聚集,并与肿瘤的分期进展相关,可能与肿瘤微环境分泌的趋化因子作用有关。     

Tpl-2表达与结直肠癌发生发展的关系及意义

目的: 研究Tpl-2在结直肠癌旁正常黏膜、结直肠腺瘤以及结直肠腺癌中的蛋白表达情况,及其与结直肠癌临床病理参数的关系,探讨Tpl-2在结直肠癌发生发展中的意义。方法: 使用组织芯片及免疫组化方法检测Tpl-2在结直肠正常黏膜、腺瘤、癌组织中的表达情况,分析Tpl-2表达与结直肠癌的临床病理参数的相关性。结果: 24例癌旁正常黏膜中,Tpl-2低表达20例(83.3%)、高表达为4例(16.7%);24例腺瘤组织中Tpl-2低表达17例(70.8%)、高表达7例(29.2%);96例癌组织中Tpl-2低表达31例(32.3%)、高表达65例(67.7%)。与正常和腺瘤比较,癌组织Tpl-2表达均明显增高(P<0.01)。腺瘤和正常黏膜中Tpl-2表达无显著差异(P>0.05)。在96例结直肠癌组织中,Tpl-2表达与N分期、TNM分期相关(P<0.05),与性别、年龄、身高体重指数(BMI)、肿瘤大小、分化程度、T分期、M分期以及K-ras基因突变状况无明显相关(P>0.05)。结论: Tpl-2与结直肠癌发生及疾病进展具有较密切的关系,可能是一个促癌因子。     

结直肠癌Stat3表达与细胞增殖、凋亡及预后的关系

目的探讨结直肠癌Stat3蛋白表达与细胞增殖、凋亡、临床病理特征及预后的关系。方法应用免疫组织化学及DNA末端标记(TUNEL)技术分别检测129例结直肠癌及癌旁组织芯片中Stat3、Cyclin D1蛋白表达及凋亡指数(AI),分析Stat3表达与临床病理特征和预后的关系及其与增殖、凋亡之间的相关性。结果结直肠癌组织中Stat3、Cyclin D1阳性表达率分别为67.4%和79.8%,均高于癌旁组织的43.4%和5.4%(P0.05),结直肠癌组织AI(5.62)低于癌旁(7.86)(P0.05)。Stat3的表达与结直肠癌的Dukes分期、淋巴结转移、肿瘤发生部位明显相关(P0.05)。结直肠癌组织中Stat3和Cyclin D1的表达呈正相关(P0.05)。Kaplan-Meier生存分析显示Stat3、Dukes分期、淋巴结转移及凋亡指数与生存期相关(P0.05);Cox回归分析显示Dukes分期为独立的风险预后因子。结论 Stat3可能参与了结直肠癌的发生发展,检测Stat3可作为评估结直肠癌患者预后及淋巴结转移的指标。     

SDF-1/CXCR4及VEGF在结直肠腺癌中的表达及其意义

目的:研究VEGF、SDF-1及CXCR4在结直肠腺癌中的表达及其与临床病理学参数之间的相关性;探讨SDF-1/CXCR4轴与VEGF在结直肠腺癌发生发展浸润转移中的作用。方法:利用实时荧光定量PCR(RT-q PCR)法及免疫组织化学SP法检测80例结直肠腺癌及其对应癌旁组织中SDF-1、CXCR4和VEGF mRNA及蛋白的表达。结果:在mRNA水平上,结直肠癌组中SDF-1、CXCR4、VEGF的表达显著高于对应的癌旁组织。免疫组化结果显示,肿瘤中c蛋白表达阳性率分别为78.7%、60%和66.3%高于对应癌旁组织的37.5%、42.5%和28.8%,差异显著(P0.05);结直肠癌中SDF-1表达水平与肿瘤细胞淋巴结转移、远处转移及TNM分期密切相关(P0.05),与患者的年龄、性别、肿瘤的大小、浸润深度等无关;CXCR4的表达与患者的淋巴结转移及TNM分期密切相关;SDF-1的表达与CXCR4及VEGF的表达成显著正相关。结论:SDF-1/CXCR4及VEGF在结直肠癌组织中高表达,并且与肿瘤生物学行为密切相关,提示其在肿瘤的发生发展浸润转移过程中发挥重要作用。     

Human papillomavirus DNA in adenosquamous carcinoma of the lung

AIM: To investigate the presence of human papillomavirus (HPV) DNA in adenosquamous carcinoma of the lung--which is relatively common in Okinawa but not in mainland Japan--and examine its histological features. METHODS: Of 207 cases where primary lung cancers were surgically removed between January 1995 and June 1997 in Okinawa, 23 were adenosquamous carcinoma. HPV was detected by non-isotopic in situ hybridisation (NISH) and polymerase chain reaction (PCR) amplification with primers specific for E6 and E7 regions of the HPV genome. PCR products were analysed by Southern blotting. Immunohistochemical determination of high molecular weight cytokeratin (HMC) and involucrin was also carried out. RESULTS: 18 cases were positive for HPV DNA by PCR and NISH. HPV types 6, 11, 16, and 18 were found. Seven cases were dual positive for different types of HPV. Using NISH, HPV was also found in the squamous cell components and in neighbouring enlarged adenocarcinoma cells. The HMC and involucrin were demonstrated immunohistochemically in the same areas. CONCLUSIONS: HPV DNA was found in a high proportion (78.3%) of adenosquamous carcinomas in Okinawa, a region where HPV has previously been shown to be prevalent in squamous cell carcinoma of the lung. The adenocarcinoma cells adjacent to the squamous cell carcinoma component were enlarged and positive for HPV, HMC, and involucrin. This is thought to indicate the transition from adenocarcinoma to squamous cell carcinoma.     

Codon 201 Mutation of DCC Gene and Tumor Biologic Behavior in Human Colorectal Carcinoma

Objective To explore the relationship between a point mutation of codon 201 in deleted in colorectal carcinoma ( DCC) gene and the biological behavior of colorectal carcinoma. Methods Tumor tissues and matched adjacent normal colon mucosa collected in 35 patients during surgical resection for colorectal carcinoma were analyzed. Forty normal colon mucosa tissues obtained by biopsy from patients who had neither colorectal tumor nor a family history of colorectal cancer during colonscop ic examination were used as control. Codon 201 mutatian was detected with allele-specific PCR and a restriction enzyme digestion method. The tumors were reviewed as clinical data, tumor location, histology,metastasis, and pathological staging (Dukes classification). Results The frequency of mutation at codon 201 in tumor tissue and corresponding adjacent normal mucosa was 71.4 % and 60 %, respectively, and either of the rates was significantly higher than that of normal control(32.5 % ). The point mutation rate in tumor tissues did not differ from that in the corresponding normal adjacent tissues. Statistic analysis showed that the mutation rate had no relationship to the sex, age of the patients, the histological pattern , differentiation, and invasion depth of the tumors. However, 93. 8 % of the mutation rate in colorectal cancer with lymph node invasion and/or distant metastasis is significantly higher than 52. 6 % of mutant rate in colorectal cancer uithout lymph nodes invasion or metastasis ( P <0. 05). Conclusion The point mutation at codon 201 of DCC gene is an early genetic event in colorectal cancer, and play some role in invasion and metastasis of colorectal carcinoma. It may serve as a useful genetic marker for identifying higher risk patients with colorectal carcinoma.     

人乳头瘤病毒16型转换基因的序列分析

为了解中国地区宫颈癌病人中人乳头瘤病毒１６型Ｅ６Ｅ７基因结构特点,从中国山东地区宫颈癌活检组织中提取组织ＤＮＡ,经ＨＰＶ多重引物ＰＣＲ法鉴定标本中感染ＨＰＶ型别,选单纯感染ＨＰＶ１６两例标本ＤＮＡ为模板进行ＰＣＲ扩增,获得ＨＰＶ１６Ｅ６Ｅ７基因后,重组入ｐＡＬＴＥＲ－１载体,进行双向测序、分析。ＤＮＡ序列分析表明：两例标本的ＨＰＶ１６Ｅ６Ｅ７序列全长均为７７６ｂｐ,与已发表的德国标准株长度相等,两     

宫颈癌组织中人乳头瘤病毒16型E7蛋白致癌机理初探

目的 研究宫颈癌组织中人乳头瘤病毒（ＨＰＶ）１６－Ｅ７蛋白对视网膜母细胞瘤基因（Ｒｅｔｉｎｏｂｌａｓｔｏｍａ）Ｒｂ蛋白及Ｅ２Ｆ－１的作用的机制,探讨ＨＰＶ１６－Ｅ７蛋白与宫颈癌发生的关系。方法 采用聚合酶链反应检测宫颈癌及正常宫颈组织中ＨＰＶ１６感染等,用蛋白印迹技术对ＨＰＶ１６ ＤＮＡ阳性的宫颈癌组织中是否存在ＨＰＶ１６－Ｅ７蛋白和Ｒ６蛋白－Ｅ２Ｆ－１形成的复合物进行检测。正常宫颈组织作为对照,     

Prevalence of human papillomavirus DNA in various histological subtypes of cervical adenocarcinoma: a population-based study.

The role of human papilloma virus (HPV) infection in the development of cervical carcinoma is well established, however, the prevalence of HPV DNA in cervical adenocarcinoma varies from study to study. It appears to be caused by a number of factors, one of which is that cervical adenocarcinomas comprise a heterogeneous group of multiple subtypes. To clarify the impact of HPV infection on the development of cervical adenocarcinoma with diverse histological subtypes, we performed a population-based study in Korean women from 15 different institutes for the status of HPV infection in adenocarcinoma of uterine cervix. A total of 432 cervical adenocarcinomas from 1997 to 2001 were reviewed and classified according to the modified WHO classification. For 135 cases, HPV typing was performed with HPV DNA chip (82 cases) and PCR HPV typing (53 cases), using formalin-fixed, paraffin-embedded archival tissue. The overall prevalence of HPV infection in cervical adenocarcinoma was 90%. The infection of HPV 16 and/or HPV 18 accounted for 78% of HPV-positive adenocarcinomas. Multiple HPV types were found in 13% of the cases. The HPV DNA was rarely detected in minimal deviation adenocarcinoma. Interestingly, HPV 16 was a predominant type in endometrioid and villoglandular types, whereas HPV 16 and HPV 18 were detected with equal prevalence in other subtypes. In conclusion, HPV infection, mostly HPV 16 and HPV 18, is highly associated with most of the cervical adenocarcinomas, whereas endometrioid and villoglandular type have a different pattern of HPV infection status. Minimal deviation adenocarcinoma does not seem to be related with HPV infection.     

人喉癌组织中人乳头瘤病毒DNA的检测

目的为探讨喉癌与人乳头瘤病毒（ＨＰＶ）感染的关系和ＨＰＶ在喉癌中基因组型的分布与表达。方法应用聚合酶链反应技术（ＰＣＲ）制备非放射性探针标记物－地高辛标记ＨＰＶ共有引物探针,对１４６例喉不同病变的新鲜组织标本（喉癌６８例,喉其它病变４８例,正常喉组织３０例）,进行ＨＰＶ６,１１,１６,１８,３１,３３,３５,４２,５８共９型ＨＰＶＤＮＡ感染的检测;阳性者用多重引物ＰＣＲ方法分型。结果喉癌ＨＰＶ感染阳性率４５．６％（３１／６８）,喉癌颈转移淋巴结组织阳性率２０．０％（３／１５）,喉癌前病变阳性率１１．８％（２／１７）,声带息肉阳性率６．３％（１／１６）,１５例癌旁及１５例癌周正常喉组织均为ＨＰＶＤＮＡ阴性。ＨＰＶＤＮＡ型别分布在喉癌中以ＨＰＶ１６、１８型为主,喉良性病变中以ＨＰＶ６、１１型为主。结论喉癌发生与ＨＰＶ感染有关。     

子宫颈癌及癌前病变HPV16、Ki-67的表达及其相关性

目的对子宫颈癌及癌前病变中HPV感染和增殖细胞核抗原Ki-67表达情况进行研究。方法对45例子宫颈浸润性鳞状细胞癌、5例子宫颈腺癌、35例子宫颈上皮内瘤变（CIN）和5例正常宫颈组织采用免疫组化EnVision法进行HPV16和Ki-67表达的检测。结果HPV16阳性率为78．9％（71／90），其中HPV16阳性率在子宫颈癌组织中为76．0％（38／50），在CIN病变中为94．3％（33／35），在正常子宫颈黏膜上皮组织中为阴性（0／5）。Ki-67阳性细胞在正常子宫颈、CIN和子宫颈癌组织中表达逐级增加，显示Ki-67表达程度与组织学类型有关。HPV和Ki-67在CIN和癌组织中的表达呈显著相关（P〈0．005）。结论瑞安地区子宫颈癌及癌前病变组织中存在HPV感染，HPV感染可能在子宫颈癌的发生、发展中起着重要作用。联合应用Ki-67与HPV可作为筛选子宫颈癌高危个体有价值的生物学标记。     

结直肠癌nm23基因表达、突变与侵袭、转移的相关性

目的　探讨nm2 3基因异常表达及突变与结直肠癌侵袭、转移之间的关系。方法　应用免疫组化LDP法、流式细胞术、PCR SSCP法对 1 5 0例结直肠癌标本及正常黏膜组织和 1 6例大肠腺瘤分 3组进行nm2 3蛋白检测和基因突变的筛查。结果　①免疫组化发现 73例结直肠癌中nm2 3 H1蛋白表达水平低于正常黏膜组织和良性腺瘤 (P <0 0 0 1 ,P <0 0 0 1 ) ,转移癌表达低于原发癌 (P <0 0 0 1 ) ,结直肠癌nm2 3 H1蛋白表达与组织分化、肠壁浸润、Dukes分期、转移均密切相关 (P <0 0 0 1 ) ,低表达组术后预后差于高表达组 (P <0 0 0 1 ) ;②流式细胞术研究表明 ,nm2 3 H1蛋白在癌组织表达低于癌旁组织 ;在结直肠癌伴有转移 (3/ 1 4 )时其表达低于未发生转移者 (1 1 / 1 4 ) ;③nm2 3 H1基因第 2～ 5位外显子在 6 3例结直肠癌组织中未发现异常突变。结论　结直肠癌中nm2 3基因突变可能是小概率事件 ,在结直肠癌的侵袭、转移中不发挥主要作用 ,而nm2 3 H1蛋白表达降低则与结直肠癌的侵袭、转移有关 ,提示检测nm2 3蛋白表达是判断结直肠癌预后有价值的指标之一。