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目的应用逆转录聚合酶链反应(RT-PCR)方法检测胃癌淋巴结微转移，探讨淋巴结微转移与胃癌生物学行为的关系。方法对中山大学附属第一医院2003年12月至2004年4月间行手术切除的30例胃癌共850枚淋巴结采用CK-20mRNA RT-PCR扩增检测微转移，分析微转移的临床病理特点。结果14例(46．7％)77枚(12．5％)淋巴结检出淋巴结微转移。弥漫型胃癌的微转移阳性率为63．2％(12／19)，明显高于肠型胃癌的18．2％(2／11，P＜0．05)。肿瘤浸润深度越深，微转移的检出率越高。结论CK-20mRNA RT-PCR法可以检测出常规病理学检查遗漏的淋巴结微转移，可显著提高淋巴结转移的检出率。淋巴结微转移与Lauren分型和浸润深度密切相关。     

IHC和RT-PCR法对检测胃癌区域淋巴结微转移的价值及其意义

目的 探讨免疫组织化学方法(IHC)和逆转录聚合酶链反应(RT-PCR)两种方法对检测胃癌区域淋巴结微转移的临床价值及意义.方法 对85例胃癌根治性手术切除的淋巴结转移患者的临床资料进行了回顾性分析.共切取淋巴结1835枚,每例平均切除21.7枚.采用IHC和RT-PCR法检测细胞角蛋白20(CK20)的表达,研究淋巴结微转移与临床病理参数和预后的关系.结果 患者的淋巴结转移率经IHC法和RT-PCR法检测从HE染色的75.3%分别上升为83.5%和90.6%.经IHC和RT-PCR法检测重新分期率分别为18.8%和37.6%.淋巴结微转移的发生与肿瘤大小、部位无关·与肿瘤Lauren分型和浸润深度密切相关.82例完成随访,平均随访时间为21.2个月.微转移与预后无明显关系.结论 IHC和RT-PCR法是检测胃癌淋巴结微转移的有效手段,能准确判断临未分期,可为制定治疗方案提供依据.     

中下段直肠癌预后与淋巴结微转移的关系

目的 观察淋巴结微转移对中下段直肠癌预后的影响.方法 应用CK-20免疫组织化学技术对56例中下段直肠癌患者共计661枚淋巴结检测微转移.结果 20例(35.7%)67枚(10.1%)淋巴结检出微转移.20例检出淋巴结微转移者中10例TNM分期提高:Ⅰ→ⅢA 3例,Ⅰ→ⅢC 2例,ⅡA→ⅢB 3例,ⅢA→ⅢC 2例.Kaplan-Meier生存分析显示,淋巴结微转移阳性患者半数生存期为(36.90±3.37)个月(95%置信区间:30.29～43.51个月),明显短于淋巴结微转移阴性者的(48.72±2.25)个月(95%置信区间:44.30～53.14个月),两者差异有统计学意义(P＜0.05).结论 中下段直肠癌淋巴结微转移检测有助于更准确地进行临床病理分期.淋巴结微转移阳性者预后较差.     

胃癌淋巴结微转移与E-钙黏附素表达的关系

目的检测E-钙黏附素(E-cadherin)在胃癌组织中的表达及探讨其与淋巴结微转移的关系.方法对30例胃癌共850枚淋巴结采用细胞角蛋白-20(Cytokerarin-20,CK-20)逆转录聚合酶链反应(RT-PCR)扩增检测微转移,采用免疫组织化学染色检测该30例胃癌组织中E-cadherin的表达情况.结果46.7%(14/30)的胃癌组织E-Cadherin表达阴性;E-Cadherin表达阴性与Lau-ren分型、淋巴结转移密切相关(P＜0.05),与分化程度及淋巴管侵犯明显相关(P＜0.001),但与性别、年龄、肿瘤位置、直径及浸润深度无关(P＞0.05).14例检测出淋巴结微转移的癌组织中10例(71.4%)E-Cadherin表达阴性,而16例未检测出淋巴结微转移的癌组织中仅有4例(25%)E-Cad-herin表达阴性,P=0.026.结论E-cadherin表达减弱或消失参与了胃癌淋巴结微转移的发生.     

结直肠癌前哨淋巴结微转移免疫组化检测的临床意义

目的探讨Cytokeratin-18(CK-18)免疫组织化学在结直肠癌前哨淋巴结转移检测中的应用价值。方法自2003年5月至2004年3月采用CK-18免疫组织化学方法检测99例SLN组织微转移癌。结果常规病理检测发现结直肠癌淋巴结转移45例,未转移54例。99例SLN常规病理癌转移阴性81例,其中CK-18免疫组织化学检测发现微转移31例。CK-18染色将15%(8/54)常规病理Dukes’A及B期患者肿瘤分期提升为Dukes’C期。CK-18免疫组织化学判定结直肠癌淋巴结转移的敏感性为98%,特异性为96%,准确度为97%。结论CK-18免疫组织化学检测结直肠癌SLN有助于提高Dukes’A及B期结直肠癌的病理分期。     

早期胃癌淋巴结微转移的临床研究

目的 探讨早期胃癌的病理及淋巴结微转移情况.方法 收集本院2005年1月至2006年12月之间共249例胃癌手术切除标本中的36例早期胃癌病例,用常规病理学方法(HE染色)及免疫组化方法(CK染色)对497个淋巴结(平均每例13.8个淋巴结)标本进行回顾性临床分析.结果 常规病理学方法检测发现4个病例共6枚淋巴结出现转移,而免疫组化方法发现8个病例(包括HE染色发现的4例)共20枚淋巴结出现转移,并且8例淋巴结微转移阳性的病例中2例癌组织位于黏膜层,6例癌组织侵及黏膜下层,提示早期胃癌淋巴结微转移与肿瘤浸润深度有关(P<0.05),并且免疫组化方法对淋巴结微转移的检出率高于常规HE染色(4.02%与1.21%;P<0.05).结论 免疫组化(CK染色)方法检测淋巴结微转移优于常规病理检查.肿瘤侵犯黏膜下层的早期胃癌更易发生淋巴结转移.     

应用RT-PCR并Southern杂交技术检测乳腺癌骨髓微转移

目的检测原发性乳腺癌骨髓微转移的发生及与其它临床参考指标的关系.方法应用逆转录-聚合酶链反应(RT-PCR)并Southern杂交技术,检测骨髓单个核细胞中细胞角蛋白19(CK-19)基因表达.结果52例骨髓标本共检出微转移19例(36.5%),其中17例淋巴结有转移者,9例CK-19表达阳性(52.9%);而无淋巴结转移的35例中,10例CK-19表达阳性(28.6%).微转移检出率与肿瘤大小有关(P＜0.05).结论以CK-19为标志物,RT-PCR并Southern杂交方法检测原发性乳腺癌骨髓微转移灵敏、特异,可做为临床判断预后的参考指标.     

中下段直肠癌E-钙黏附素表达与淋巴结微转移的关系研究

目的:分析中下段直肠癌E-钙黏附素(E-cadherin)表达与淋巴结微转移的关系。方法:应用CK-20免疫组化技术,对56例中下段直肠癌中661枚淋巴结微转移状况进行检测,同时观察肿瘤组织中E-钙黏附素的表达情况。结果:HE染色检测出29例中的190枚淋巴结呈转移,其CK-20免疫组化检测均呈阳性,后者在该29例中另检出12例55枚淋巴结呈阳性;在27例HE染色未检出淋巴结转移者中,有8例12枚淋巴结免疫组化检测呈阳性。20例(36%)67枚(10%)淋巴结检出微转移。中下段直肠癌E-钙黏附素表达阴性率为44.6%(25/56)。中下段直肠癌E-钙黏附素表达阴性与浸润深度(P=0.028)、分化程度(P=0.012)和淋巴结转移(P=0.007)密切相关。20例检测出淋巴结微转移的癌组织中14例(70%)E-钙黏附素表达阴性,而36例未检测出淋巴结微转移的癌组织中仅有11例(30.6%)E-钙黏附素表达阴性;二者差异有统计学意义(P=0.004)。结论:中下段直肠癌E-钙黏附素表达下调参与了淋巴结微转移的发生。     

荧光定量PCR检测pNo期结直肠癌淋巴结中CK20mRNA的表达及其临床意义

目的 探讨N0期大肠癌淋巴结微转移的检测及其临床意义.方法 采用荧光定量PCR方法 检测62例行根治术的N0期大肠癌患者的548枚淋巴结中细胞角蛋白20(cytokeratin,CK20)mRNA的表达.结果 本组62例N0期大肠癌患者的548枚淋巴结中,有24例(39%)共55枚(10.0%)淋巴结中检出微转移.其中第Ⅰ、第Ⅱ、第Ⅲ组淋巴结微转移率分别是15.8%、5.0%和3.3%.微转移与患者年龄、性别、肿瘤大小、肿瘤部位和分化程度等尤关,但与肿瘤浸润深度相关(x2=5.462,P<0.05).结论 检测N0期大肠癌淋巴结中CK20mRNA的表达可对精确临床分期、了解淋巴结微转移分布及制定合理的治疗方案提供一定的理论依据.     

甲状腺滤泡状癌淋巴结微转移与中央组淋巴结处理的探讨

目的 从淋巴结微转移的角度探讨甲状腺滤泡状癌中央组(Ⅵ组)淋巴结处理的意义.方法 选择细胞角蛋白(CK-19)单克隆抗体,应用免疫组化SP法对68例326枚常规病理检查阴性的颈淋巴结(pN0)进行检测,确定其微转移情况并与临床及随访资料进行对比分析.结果 326枚pN0淋巴结中微转移阳性46枚,包括Ⅱ组4枚(4/42),Ⅲ组5枚(5/34),Ⅳ组5枚(5/49),Ⅴ组1枚(1/17),Ⅵ组31枚(31/184).在淋巴结微转移阳性的14例中6例(6/14)出现局部复发或远处转移,而54例微转移阴性病人中仅3例(3/54)复发或远处转移(P＜0.01).结论 甲状腺滤泡状癌Ⅵ组淋巴结是容易发生微转移的区域淋巴结;且淋巴结微转移与肿瘤复发及转移有密切关系.甲状腺滤泡状癌手术时应常规清扫Ⅵ组淋巴结.     

胃癌淋巴管生成、淋巴管浸润及淋巴结微转移的临床意义

目的探讨胃癌淋巴管生成、淋巴管浸润及淋巴结微转移的临床意义。方法免疫组化法检测68例胃癌原发灶中D2-40的表达及其中51例胃癌的791枚淋巴结中CK20和CKpan的表达，结合患者的l临床病理特征进行综合分析。结果胃癌HE染色淋巴管浸润（LVI-HE）和D240染色淋巴管浸润（LVI-IM）的阳性率分别为66．2％（45／68）和76．5％（52／68），差异无统计学意义（P=0．118）。LVI-IM阳性率与肿瘤浸润深度（P=0．044）、TNM分期（P=0．003）及存在淋巴结转移（P=0．000）有关。68例胃癌平均淋巴管密度（LVD）为（18．19&#177;7．44）个／HP.LVD升高与LVI-HE阳性（P=0．040）、LVI—IM阳性（P=0．001）、静脉浸润（P=0．037）、TNM分期较晚（P：0．020）及存在淋巴结转移（P=0．001）有关系。LVD值≥15个／HP者近期生存率较LVD值≤14个／HP者明显降低（P=0．032）。51例胃癌HE染色和CK（CK20或CKpan）染色检出淋巴结转移率分别为74．5％（38／51）和88．2％（45／51），791枚淋巴结的转移淋巴结检出率由HE染色的32．0％（253／791）提高到CK染色的41．5％（328／791），P〈0．001。CKpan的微转移检出率明显高于CK20（P=0．003）。微转移淋巴结数量与肿瘤大小（P=0．001）、LVIHE（P=0．040）、肿瘤浸润深度（P=0．018）及TNM分期（P=0．012）有关。微转移淋巴结的检出使淋巴结转移站别及TNM分期迁移：7例N0→N1，6例N1→N2，1例N2→N3；4例Ⅰb→Ⅱ，4例Ⅱ→Ⅲa，3例Ⅲa→Ⅲb，1例Ⅲb→Ⅳ。结论D2-40及CK检测在诊断淋巴管浸润和淋巴结微转移上优于HE检查。CK20和CKpan的联合检查有利于发现微转移淋巴结。肿瘤TNM分期越晚，越易发生淋巴结微转移。LVI-IM、LVD及淋巴结微转移三者都与胃癌淋巴结转移有关。LVD值较高者近期生存率较低。     

早期胃癌CD44v6的表达及与淋巴结微转移的关系

目的：研究早期胃癌组织CD44v6的表达及与淋巴结微转移的关系,并探讨其临床意义。方法：对64例早期胃癌患者手术切除的原发灶行CD44v6免疫组化检测,对清扫的常规HE染色阴性的782个淋巴结行CK20免疫组化检测。结合临床病理学指标,对CD44v6在原发灶的表达和淋巴结微转移与肿瘤直径、组织学类型及侵犯深度等的关系,以及CD44v6的表达与淋巴结微转移之间的关系进行分析。结果：64例早期胃癌患者23例CD44v6染色阳性,6例14个淋巴结内有微转移。23例CD44 v6表达阳性的患者中4例（17.39%）发生微转移,31例CD44v6表达阴性的患者中2例（6.45%）发生微转移。结论：淋巴结微转移与肿瘤直径、组织学类型及侵犯深度相关。CD44v6染色阳性与肿瘤直径和组织学类型无关,但与侵犯深度相关。原发灶CD44染色阳性与淋巴结微转移有相关性。     

Micrometastasis in lymph nodes and microinvasion of the muscularis propria in primary lesions of submucosal gastric cancer

BACKGROUND: It is important to clarify the clinicopathologic characteristics of micrometastasis in lymph nodes and microinvasion in primary lesions for the treatment options with regard to submucosal gastric cancer. METHODS: We examined 1945 lymph nodes and 68 primary tumors resected from 79 patients with submucosal gastric cancer. Two consecutive sections were prepared for simultaneous staining with ordinary hematoxylin and eosin and immunostaining with anticytokeratin antibody (CAM 5.2), respectively. RESULTS: The incidence of nodal involvement in 79 patients with submucosal gastric cancer increased from 13% (10/79 patients) by hematoxylin and eosin staining to 34% (27/79 patients) by cytokeratin immunostaining. Micrometastases in the lymph nodes were found in 17 of 69 patients (25%), with cancer-free nodes examined by hematoxylin and eosin. Microinvasion to the muscularis propria was found in 11 of 68 patients (16%) who were histologically diagnosed with submucosal gastric cancer. Survival analysis demonstrated a lesser 5-year survival in the patients with micrometastasis in lymph nodes (82%) and with microinvasion to muscularis propria (73%). A high incidence of nodal involvement was found in submucosal cancers of large size (> 2 cm; 43%), a depressed type (48%), lymphatic invasion (73%), and deeper submucosal invasion (submucosal 3, 53%). A higher incidence of microinvasion was found with the diffuse-type carcinoma (33%). CONCLUSIONS: Cytokeratin immunostaining is useful for detecting micrometastasis and microinvasion in submucosal gastric cancer. Tumor size, macroscopic type, lymphatic invasion, and the depth of submucosal invasion are strongly associated with lymph node involvement.     

Clinical impact of micrometastasis of the lymph node in gastric cancer

Micrometastasis in regional lymph nodes has been observed immunohistochemically, but the biological and clinical roles of minute nodal invasion of carcinoma in gastric cancer remain unclear. We used the anti-cytokeratin (AE1/AE3) antibody to immunohistochemically detect nodal micrometastatic lesions that could not be identified by routine pathological examination. A total of 4203 lymph nodes were examined in 180 gastric cancer patients. Lymph node metastasis was found in 36 of the 180 patients by routine pathological evaluation. Immunohistochemically micrometastasis was detected in the lymph nodes of 19 node-negative patients. Micrometastasis was not detected in any of the mucosal gastric cancer patients who underwent lymph node dissection. Gastric cancer patients with more than six metastatic lymph nodes all had nodal micrometastasis. Patients with micrometastasis had a significantly poorer survival rate than those without micrometastasis (P < 0.05). Based on the present results the presence of lymph node micrometastasis may provide a more accurate indication for surgical outcome in gastric cancer patients at the same clinical stage.     

Impact of Lymph Node Micrometastasis in Patients with Pancreatic Head Cancer

Purpose The purpose of this study was to investigate the clinical significance of nodal micrometastasis in patients who underwent a curative operation for pancreatic cancer. Experimental Design Fifty-eight patients underwent a macroscopically curative resection with extended lymph node dissection for pancreatic cancer. The total number of resected lymph nodes was 1,058, and 944 histologically negative lymph nodes were subjected to immunohistochemical staining to detect occult micrometastases. Results Nodal micrometastases were detected immunohistochemically in 147 out of 944 resected histologically negative lymph nodes (15.6%). Forty-four of all 58 patients (75.9%) and 13 of the 23 histologically node-negative patients (56.5%) had nodal micrometastases. Nodal micrometastases existed in the N1 lymph node area most frequently, followed by the N2 and N3 lymph node areas. The distribution was similar to that of histologically metastatic lymph nodes. Ten out of 16 patients (62.5%) with histological N1, and 5 out of 16 patients (31.3%) with histological N2 had nodal micrometastases beyond the histological lymph node status. Three and 5-year survival rates of pN0 patients without lymph node nodal micrometastases were both 60.0%, while those with nodal micrometastases were 19.2% and 0%, respectively. There was statistically significant difference between the both groups (P = 0.041). Conclusions Nodal micrometastasis in pancreatic cancer existed in wider and more distant areas than histological lymph node status, and it was an unfavorable predictive factor, even in N0 patients.     

Dukes A、B期大肠癌淋巴结微转移的检测及其对预后的影响

目的：探讨Dukes A、B期大肠癌淋巴结微转移的检测和淋巴结微转移对预后的影响.方法：于前瞻性研究31例行根治性手术的Dukes A、B期大肠癌病人,应用逆转录聚合酶链反应（RT-PCR）检测所清除的398枚淋巴结中细胞角蛋白（cvtokeratin,CK）20 mRNA的表达以检出微转移;经5年以上的随访,探讨淋巴结微转移对预后的影响和术后复发的可能原因.结果：在31例Dukes A、B期大肠癌病人的398枚淋巴结中,有15例（48.39%）共46枚（11.56%）淋巴结检出微转移.单因素分析提示微转移的淋巴结数量、位置及肿瘤生长方式与术后复发有关;Logistic多元回归模型提示,3枚以上淋巴结发生微转移与复发紧密联系.结论：CK20 RT-PCR是检测Dukes A、B期大肠癌淋巴结微转移灵敏而特异的方法.3枚以上淋巴结发现微转移是预示复发的独立因素.     

Micrometastasis to axillary lymph nodes and bone marrow in breast cancer patients

The axillary lymph nodes of 100 lymph node-negative breast cancer patients with known bone marrow status have been re-examined to explore the presence of micrometastasis in lymph nodes and the covariance of micrometastasis to bone marrow and lymph nodes. Nodes were serially sectioned at three intervals of 100 microm, followed by immunohistological (two sections) and haematoxylin-eosin staining (one section). Tumours were mainly T1 and T2, and the patients had on average 13 (4-22) lymph nodes removed. In two patients, micrometastasis was detected in one node. Another 25 patients possessed single positive immunostained cells mimicking tumour cells. These cells have been shown to be false positive cells by Perl and melanin staining. One patient had metastasis to several nodes missed by the original examination. Immunocytochemical detection of micrometastasis in bone marrow revealed 11 marrow-positive patients. This study has identified a low frequency of micrometastasis to lymph nodes, and no covariance with micrometastasis in the bone marrow was seen. Bone marrow micrometastasis may be an independent prognostic variable, separate from axillary node status.     

淋巴结微转移检测对胃癌病理分期的影响

目的 分析淋巴结微转移对胃癌pN分期的影响。方法 采用CK-20 mRNA逆转录聚合酶链反应（RT-PCR）技术对2003年12月至2004年4月间行手术切除的30例胃癌患者共计850枚淋巴结扩增检测微转移。结果 应用苏木精-伊红染色法淋巴结转移检出率为27．1％（233／850），而RT-PCR法淋巴结转移的检出率则为36．5％（310／850）；两种方法比较，P〈0．01，差异有统计学意义。77枚淋巴结检出有微转移（12．5％，77／617）。有7例（23．3％）患者的肿瘤TNM分期提高，分别为IB→Ⅱ、IB→ⅢA、Ⅱ→ⅢA、ⅢA→ⅢB、ⅢA→Ⅳ各1例，ⅢB→Ⅳ2例。结论 RT-PCR法可以显著提高淋巴结转移的检出率，有助于更准确地进行临床病理分期。     

早期胃癌直径、淋巴结微转移与组织MMP-7表达强度的相关性

目的:研究早期胃癌直径、淋巴结微转移与基质金属蛋白酶-7表达的相关性.方法:55例早期胃癌经黏膜颜色、术后病理证实为早期胃癌.用免疫组织化学法检测MMP-7的表达强度和淋巴结微转移.结果:直径大于20mm早期胃癌的MMP-7表达阳性率明显高于直径小于10 mm的病例(P＜0.045),亦高于直径10～20 mm病例的表达,但无统计学意义(P＞0.104).MMP-7的表达与胃癌的组织分型明显相关,低分化癌MMP-7的表达高于高分化癌(P＜0.015),亦高于中分化癌的表达,但无统计学意义.直径大于20mm早期胃癌的淋巴结转移率高于直径10～20mm的淋巴结转移率.结论:早期胃癌直径、淋巴结微转移与基质金属蛋白酶-7的表达相关,直径大于20mm患者MMP-7表达强阳性率和淋巴结转移率高.     

术中美蓝染色及抗细胞角蛋白20检测淋巴结及其微转移在直肠癌患者中的临床应用

目的探讨术中经直肠上动脉注射美蓝行直肠系膜及淋巴结染色检测淋巴结及其微转移方法的临床意义。方法对30例行根治性手术治疗的直肠癌患者,术中经直肠上动脉插管注射美蓝6ml(美蓝染色组),使直肠系膜及各组淋巴结染色以统计淋巴结数目,同时行抗细胞角蛋白20(CK20)免疫组织化学(免疫组化)染色,检测其微转移情况;并与同期32例行直肠癌根治术患者术中常规手检法(常规手检组)检出的淋巴结数目进行对比。结果美蓝染色组肠旁淋巴结、肠系膜淋巴结、肠系膜根部淋巴结检出数目均明显多于常规手检组(P均=0.000)。美蓝染色组淋巴结常规病理检查阴性者,经CK20免疫组化染色,其转移阳性率增加17.7%;14例DukesB期患者(46.7%)经淋巴结CK20免疫组化染色需重新确定为DukesC期,11例患者(36.7%)出现跳跃淋巴结微转移现象。结论术中经直肠上动脉注射美蓝行淋巴结染色能提高淋巴结检出率。对常规病理检查阴性的淋巴结行CK20免疫组化染色检查其微转移灶,能提高淋巴结阳性检出率,使病理分期更准确。