结直肠癌CD44V6、E-cadherin和VEGF的表达及其意义

目的：探讨结直肠癌CD44V6、E-cadherin和VEGF的表达及其临床意义。方法：应用免疫组化S－P法检测46例结直肠癌CD44V6、E-cadherin和VEGF的表达，并分析其与结直肠癌临床病理特征的关系。同时检测了23例淋巴结转移灶CD44V6、E-cadherin和VEGF的表达。结果：结直肠癌组织中CD44V6、E-cadherin和VEGF阳性表达率分别为50．0％、56．5％、和50．0％；原发灶CCD4V6和VEGF阳性表达率低于淋巴结转移灶（60．9％和56．％），而原发灶E-cadherin阳性表达率则高于淋巴结转移灶（43．5％）。结直肠癌CDV46和VEGF阳性表达与淋巴结转移，局部浸润深度和Dukes分期等有关。E-cadherin阳性表达与结直肠癌临床病理特征无关。结论：CD44V6和VEGF是结直肠癌发生与发展重要的促进因子，其过度表达与结直肠癌浸润转移，病理分期有关，可作为预测结直肠癌预后的生物学指标之一。     

Ⅰ期结直肠癌淋巴血管侵犯的免疫组化分析:与预后及淋巴结微转移之间关系

部分研究表明隐匿性淋巴结转移(微转移)与Ⅰ期结直肠癌预后不良有关。因此,作者分析了一组Ⅰ期结直肠癌中淋巴结微转移与淋巴血管侵犯(LVI)或淋巴血管密度(LVD)之间的关系,所有病例均有随访,有无特征性描述疾病级数的病例均包含在内。LVD与LVI可通过免疫组化法检测淋巴血管特异性标记物D2-40来测量。在已经有微转     

Raptor、Rictor与结直肠癌血管生成的相关性及其临床意义

目的检测Raptor、Rictor与血管生成相关因子HIF-1α、HIF-2α以及VEGF在结直肠癌中的表达,探讨Raptor、Rictor与结直肠癌血管生成的相关性及其临床意义。方法采用免疫组化、Western blot、RT-PCR法检测120例结直肠癌组织及60例正常结直肠黏膜组织中Raptor、Rictor及HIF-1α、HIF-2α、VEGF的表达及其差异;CD34标记微血管密度(microvascular density,MVD);分析各指标间的相关性以及与结直肠癌临床病理特征的关系。结果 Raptor、Rictor、HIF-1α、HIF-2α和VEGF蛋白在结直肠癌中的阳性率均明显高于正常结直肠黏膜组织(P0.05)。Raptor、Rictor在中、低分化结直肠癌中的表达高于高分化结直肠癌(P0.05),同时在淋巴结转移组的表达高于无淋巴结转移组(P0.05);HIF-1α、HIF-2α和VEGF在淋巴结转移组的表达高于无淋巴结转移组(P0.05);Raptor、Rictor阳性的结直肠癌组织中MVD明显高于Raptor、Rictor阴性的结直肠癌组织(P0.05);Raptor与HIF-1α、VEGF在结直肠癌中表达呈正相关(P0.01),Rictor与HIF-2α、VEGF表达呈正相关(P0.01),Raptor与Rictor在结直肠癌中表达呈负相关(P0.01)。结论 m TOR核心分子Raptor、Rictor与结直肠癌的发生、发展及血管生成密切相关,两者以不同途径协同促进结直肠癌的血管生成。     

结直肠癌淋巴管生成的特点及其临床病理意义

目的探讨结直肠癌中淋巴管的分布特点、增殖状态及其与转移和预后的关系。方法采用淋巴管特异标记podoplanin对96例结直肠癌及其相应正常组织进行免疫组织化学染色检测微淋巴管密度,以CD34标记血管检测微血管密度作为对比,并分别与Ki-67进行双标免疫组织化学染色检测淋巴管和血管增殖活性,结合结直肠癌临床病理参数和预后分析。结果结直肠癌中心及浅表部淋巴管多为闭锁的条索状,边缘区淋巴管多呈管样扩张状。结直肠癌边缘区淋巴管密度(51.2±25.5)及较正常结直肠组织(29.4±9.0)和肿瘤其他区域显著性增高(P<0.01),并且其淋巴管内皮Ki67指数(0.23±0.17)也较其他区域显著性增高(P<0.05)。结直肠癌边缘区微淋巴管密度与淋巴管受累、淋巴结转移、远处器官转移及预后密切相关(P<0.01或P<0.05)。结论结直肠癌组织中存在新生淋巴管,且主要分布于肿瘤边缘区,癌周围淋巴管密度增加与癌细胞转移相关,结直肠癌边缘区微淋巴管密度测定对评估其淋巴结转移和预后判断可能具有意义。     

TIMP-3基因甲基化与结直肠癌临床病理的关系

目的：探讨TIMP-3基因甲基化与结直肠癌临床病理指标和转移复发的关系。 方法： 采用巢式甲基化特异性PCR技术（nMSP法）检测100例结直肠癌组织和100例癌旁非癌组织TIMP-3基因甲基化；采用RT-PCR检测100例结直肠癌组织和100例癌旁非癌组织TIMP-3 mRNA的表达。 结果： 肿瘤组织TIMP-3 mRNA的表达阳性率为64%，肿瘤组织TIMP-3 mRNA的表达率明显低于癌旁非癌组织（P＜0.01）；TIMP-3 mRNA的表达率无淋巴结转移组（34/42）高于淋巴结转移组（30/58）（P＜0.01），甲基化阳性率Duke’s C+D期伴淋巴结转移组明显高于Duke’s A+B期不伴淋巴结转移组（P＜0.05）。结肠近端、分化程度差的结直肠癌组织甲基化阳性率明显高于远端直肠和分化程度高者（P＜0.05）。 结论： TIMP-3基因甲基化容易发生在结肠近端、Duke’s C、D期、伴淋巴结转移、细胞分化差和浸润型结直肠癌患者。     

直肠系膜淋巴结分布特点及其临床意义

目的：通过研究尸体及活体术后标本观察直肠系膜淋巴结的数目及分布，探讨直肠癌全直肠系膜切除术后患者的复发和转移之间的关系。方法：30例福尔马林固定的尸体和102例活体术后标本解剖，自直肠上动脉开始沿其最小的分支至肛门直肠环水平进行解剖并登记。结果：30例尸体标本直肠系膜共发现淋巴结248枚，平均8．3枚(标准差4．41)，淋巴结大小为1．8～10mm。102例行直肠癌全直肠系膜切除术的病例共发现淋巴结916枚淋巴结，平均8．98枚。所有淋巴结均送病理，其中有77枚(8．41％)有癌转移。结论：通过尸体与活体标本淋巴结的解剖，大部分淋巴结分布在直肠系膜周围靠近腹膜返折处。淋巴结数目变化范围可指导直肠癌全直肠系膜切除术后标本的病理解剖及指导术后病人的治疗。     

EGFR和Ki-67在肠癌组织中的表达及临床意义

目的:探讨表皮生长因子受体(epidermal growth factor receptor,EGFR)和Ki-67在结直肠癌(colorectal cancer,CRC)组织中的表达及临床意义.方法:应用免疫组织化学染色检测100例CRC组织中EGFR和Ki-67蛋白的表达,并分析其与临床病理特征的关系.结果:EGFR,Ki-67在CRC中的阳性表达率分别为70.0％,85.0％.EGFR的阳性表达在结直肠癌的不同分化程度和TNM分期中差异有统计学意义(P＜0.05),与淋巴结转移呈正相关(P＜0.05).Ki-67的表达与结直肠癌组织分化程度、淋巴结转移和肿瘤原发部位有关(P＜0.05).在CRC组织中,Ki-67与EGFR蛋白呈显著正相关(r=0.581,P＜0.001).结论:EGFR和Ki-67蛋白的表达与组织分化程度及淋巴结转移密切相关,二者的联合检测有助于结直肠癌恶性程度的判断和预后的估计.     

结直肠癌原发灶及淋巴结转移灶中SATB2表达及意义

目的：探讨富含AT序列特异性结合蛋白2(special AT-rich sequence-binding protein 2, SATB2)在结直肠癌组织及对应淋巴结转移灶中的表达,并分析其与临床病理特征的关系。方法应用免疫组化EnVision法检测200例结直肠癌原发灶及80例淋巴结转移灶中SATB2蛋白表达。结果结直肠癌原发灶中SATB2高表达率为25．0%(50/200),中等强度表达率为36．0%(72/200),阴性率为39．0%(78/200)。对应淋巴结转移灶中SATB2高表达率为15．0%(12/80),中等强度表达率为28．8%(23/80),阴性率为56．2%(45/80)。 SATB2在结直肠癌原发灶中的表达与肿瘤大小、分化程度,浸润深度、淋巴结转移以及TNM分期显著相关(P<0．05),与患者年龄、性别及远处转移无相关性。 SATB2在结直肠癌淋巴结转移灶中表达显著低于原发灶(P<0．05),其在转移灶中的表达与临床病理因素无明显相关性。结论 SATB2低表达与结直肠癌的发生、发展相关,在肿瘤转移过程中表达明显降低,有望成为新型的结直肠癌分子靶标。     

Lumican基因在结直肠癌中表达的相关研究

目的探讨Lumican基因的表达与结直肠癌发生、发展之间的关系。方法应用RT- PCR方法检测49例结直肠癌组织及其癌旁正常黏膜上皮组织内Lumican mRNA的表达水平,并进行统计学分析。结果RT-PCR结果表明结直肠癌组织中Lumican mRNA均呈阳性表达,癌旁正常黏膜上皮组织内Lumican mRNA的表达显著低于癌组织(P<0.01)。Lumican mRNA的表达水平与患者的性别、年龄、组织病理分型、淋巴结转移及Dukes分期无关。结论结直肠癌组织中Lumican mRNA的表达增强提示Lumican mRNA在结直肠癌组织中的高表达对结直肠癌的发生发展可能起重要作用。     

结直肠癌组织PSMA标记的微血管密度与相关临床病理特征的关系

目的 探讨结直肠癌组织中PSMA标记的微血管密度情况，并分析其与临床病理学特征及预后的相关性。方法 采用免疫组化EnVision法检测102例结直肠癌组织中PSMA标记的微血管密度情况，PCR法检测结直肠癌组织中微卫星稳定和KRAS基因突变情况，分析PSMA微血管密度与临床病理特征及预后的相关性。结果 PSMA均明确表达于肿瘤间质新生的血管内皮细胞膜，在肠壁组织的正常血管内皮未见表达；PSMA标记的微血管密度平均值为12.11,其在肿瘤组织及正常对照组织中的表达差异有统计学意义(P<0.001);PSMA标记的微血管密度与结直肠癌组织学类型(P=0.030)和淋巴结转移(P=0.033)相关。结论 PSMA在结直肠癌肿瘤相关血管中表达较高，且与组织学分型、淋巴结转移显著相关，提示PSMA可能作为结直肠癌血管靶向治疗的新靶点，为PSMA用于结直肠癌的靶向治疗提供理论依据。     

mRNA markers of breast cancer nodal metastases: comparison between mammaglobin and carcinoembryonic antigen in 248 patients

Histological detection of axillary lymph node metastases is still the most valuable prognostic parameter for breast cancer, but about 30% of node-negative patients relapse within five years, suggesting that current methods are inadequate for identifying metastatic disease. More sensitive, PCR-based methods for the detection of metastatic cells are now available, enabling the amplification of cancer cell-specific mRNA messages by the RT-PCR assay. An ideal tumour marker, consistently expressed in tumour samples and not at all in normal lymph nodes, remains to be identified. The present study first investigated the expression of seven mRNA markers, CEA, CK19, c-Met, mammaglobin, MUC-1, beta1-->GalNAc-T and p97, selected on the basis of their previously reported specificity for breast cancer cells. Eighteen lymph nodes were examined from patients without tumours. Only mammaglobin mRNA and CEA mRNA were not expressed in normal nodes. All of the other markers showed a band of expression in 17%-55% of cases, indicating that they are not breast cancer-specific. CEA mRNA and mammaglobin mRNA expression could be detected in 15/20 (75%) and 19/20 (95%) primary breast carcinomas, respectively. The expression of mammaglobin mRNA and CEA mRNA was then compared in axillary lymph nodes from 248 consecutive breast cancer patients, 89 with histologically documented lymph node metastasis and 159 without histological evidence of metastatic disease. Ninety-seven per cent of the patients with histologically involved nodes showed expression of mammaglobin mRNA, whereas CEA mRNA was expressed in 79% of these cases. In the group of patients with histologically negative lymph nodes, 46 (29%) and 32 (20%) were found to be positive for mammaglobin and CEA expression, respectively, indicating the presence of metastases not detected by routine histological examination of one lymph node section. These results show that both mammaglobin RT-PCR and CEA RT-PCR are useful tools for the detection of breast cancer metastases in axillary lymph nodes. The detection sensitivity of the mammaglobin RT-PCR is far superior to that of the CEA RT-PCR, allowing the diagnosis of occult metastases in nearly one-third of cases.     

定量RT—PCR法检测乳腺癌前哨淋巴结CK19的研究

目的探讨定量RT—PCR法检测细胞角蛋白（CK19）在乳腺癌前哨淋巴结（SLN）中的表达，提高前哨淋巴结活检中微转移的检出率。方法采用常规病理检查法（HE染色）和定量RT—PCR法检测了40例乳腺癌患者SLN的CK19的表达量，同时选取10例来源于胃肠道的良性病变淋巴结作为定量RT—PCR检测的对照组。结果CK19在良性病变的淋巴结中没有表达。常规病理检查的敏感度为42．9％（9／21），假阴性为57．1％（12／21）．准确率为70．0％（28／40）。定量RT—PCR法检测出常规病理未检出的微小转移病例12例，敏感度为95．2％（20／21），假阴性为4．8％（1／21），准确率为97．5％（39／40）。结论前哨淋巴结活检可有效判断乳腺癌腋淋巴结转移状态，应用定量RT—PCR法检测CK19在SLN中的表达，可提高敏感度及准确率。     

The development of optimal pathological assessment of sentinel lymph nodes for melanoma

1158 sentinel lymph nodes (SLNs), excised from patients with primary cutaneous melanoma, were assessed pathologically using histology with immunohistochemistry (IHC) on all nodes, and RT-PCR for Mart-1 and tyrosinase on 55 nodes. RT-PCR was compared with the histology and IHC assessed on the same nodes. The evaluation of progressively more detailed protocols for histology and IHC modulated by the RT-PCR results led to a procedure that consistently detects metastases in 34% of patients submitted to SLN biopsy for cutaneous melanomas with a vertical growth phase and a mean thickness of 2.02 mm (range 0.25, with regression, to 19 mm). As this technique is virtually free of false positives and produces only a marginally lower detection rate than RT-PCR, which was subject to false positives of 7% in our study, it is suggested that this extended protocol should be the basis on which further evaluation of the place of RT-PCR in SLN assessment takes place. The evolved protocol described here has been adopted by the EORTC as the standard procedure for pathological handling of sentinel lymph nodes for melanoma when SLN status is a criterion in their clinical trials or studies.     

Cytogenetic and immunologic phenotype findings in Hodgkin's disease

There are very few chromosome studies using banding techniques of lymph nodes in Hodgkin's disease (HD), and determinations of immunologic phenotypes are scarce. We have performed both cytogenetic and immunologic studies in 12 of 22 lymph node biopsies of different histologic types obtained from 20 HD patients (no mitotic cells were found in the remaining ten lymph nodes). A near-diploid modal number was obtained in 80% of the cases, and 20% showed a bimodal distribution. Clones were observed in 50% of HD lymph nodes, with chromosome markers in 60% of them. Markers 15q+, 5p-, and der(X) and a trisomy of chromosome #21 were observed in our cases. Seventy-one percent of the lymph nodes studied showed a predominance of T lymphocytes. Within the lymph nodes, where the karyotype was determined, 4/12 lymph nodes presented a predominance of B lymphocytes, and they were all included in the group with structural chromosome abnormalities.     

Detection of micrometastases in bone marrow and sentinel lymph nodes of breast cancer patients

Objective: To study the sensitivity and clinical significance of HE-staining,IHC and RT-PCR in detecting breast cancer micrometastases in bone marrow and sentinel lymph nodes (SLNs). Methods:After general anesthesia, all patients underwent bone marrow puncture and sentinel lymph node biopsy (SLNB) by 1% isosulfan blue, and then HE-staining,IHC and RT-PCR were used to detect micrometastases. Results:Of 62 patients with breast cancer whose axillary lymph nodes showed negative HE-staining results, 15 cases presented with positive RT-PCR and 9 cases showed positive IHC results positive in bone marrow micrometastases detection. PT-PCR and IHC showed good uniformity(kappa=0.6945)and there was significant difference in detective rate between these two methods (χ2=4.1667,P=0.0412). In SLN samples, 13 showed positive RT-PCR results, while 7 showed positive IHC results. PT-PCR and IHC showed good uniformity (kappa=0.6483)and significant difference was also found in detective rate between these two methods (χ2=4.1667,P=0.0412). Both bone marrow and SLN samples were RT-PCR positive in 3 cases,which indicated that bone marrow micrometastases did not always accompany SLN micrometastases(χ2=0.067,P=0.796). Conclusion: Even if no axillary lymph node involvement or distant metastases are present in routine preoperative examination, micrometastases can still be detected in bone marrow or SLNs. Because the bone marrow micrometastases and axillary node micrometastses are not present simultaneously, combination test of multiple indicators will detect micrometastases more accurately.