CD44基因在肠息肉和结直肠癌中的表达及其临床意义

摘 要：［目的］ 检测CD44基因在肠息肉和结直肠癌中的表达,探讨其对结直肠肿瘤癌变的临床意义。［方法］ 采用Real-time PCR方法检测CD44在82例结直肠增生型炎性息肉、腺瘤性息肉和癌变息肉组织中的表达。［结果］ 增生型炎性息肉组织中CD44 mRNA相对表达量为0.0006±0.0001,腺瘤性息肉组织中为0.0019±0.0006,癌变息肉组织中为0.0073±0.0016,三组相比,P均<0.05。有淋巴结转移者与无淋巴结转移者肿瘤组织中CD44 mRNA表达量相比,P<0.05;TNM分期Ⅲ+Ⅳ者与Ⅰ+Ⅱ者肿瘤组织中CD44 mRNA表达量相比,P<0.05。相关性分析结果显示,CD44的表达强度与肠息肉癌变进展呈正相关(r=0.562,P<0.0001)。［结论］ CD44基因在结肠癌组织中表达异常升高,提示CD44可能与结肠癌的发生、发展有密切关系。     

NOB1在结直肠癌中的表达及其临床意义

目的：研究NOB1编码蛋白在结直肠癌及正常结直肠黏膜（距癌组织边缘5cm以上）、结直肠良性息肉中的表达特征.探讨结直肠癌中NOB1表达的临床病理意义.方法：利用免疫组织化学SABC法检测87例结直肠癌组织、22例正常结直肠黏膜组织18例结直肠息肉组织中NOB1的表达.结果：NOB1在结直肠癌,结直肠良性息肉及正常结直肠黏膜中的阳性表达率分别为74.7％、44.4％、13.6％,三组进行比较,差异有统计学意义（P＜0.01）.细胞分化差的结直肠癌NOB1蛋白阳性率表达高（P＜0.05）与患者年龄,性别,肿瘤浸润深度及淋巴结转移无明显相关性.结论：NOB1蛋白在结直肠癌中表达升高,并与大肠癌临床病理学特征有关.     

结直肠癌及癌前病变中DNA倍体分析及其临床意义

目的:检测结直肠癌及癌前病变细胞内的DNA含量及DNA指数,探讨其辅助诊断和预测癌变危险度的意义.方法:应用流式细胞仪(FCM)对400例结直肠组织细胞的细胞核进行DNA倍体分析,其中正常结直肠黏膜组30例,增生性息肉组30例,绒毛状腺瘤组190例,结直肠癌组150例,检测其异倍体检出率和DI值.结果:结直肠癌组的DNA异倍体检出率显著高于正常直肠黏膜组3、增生性息肉组及绒毛状腺瘤组(P＜0.01),绒毛状腺瘤伴重度不典型增生组的DNA异倍体检出率显著高于正常直肠黏膜组3、增生性息肉组、无不典型增生的绒毛状腺瘤组及伴轻度、中度不典型增生的绒毛状腺瘤组(P＜0.01).随着结直肠癌的组织学分化程度降低,其DNA异倍体DI值呈增加趋势并有显著性差异(P＜0.01).结论:DNA含量和倍体的检测有助于了解肿瘤细胞的增殖情况及恶性程度,对诊断结直肠良恶性肿瘤、预测癌变危险度、判断预后均具有重要的参考价值.     

大肠癌组织中CD44V6、MVD的联合表达及其临床意义

目的　探讨CD44V 6、微血管密度 (MVD)在大肠癌组织中的表达及其临床意义。方法　采用免疫组织化学SP法 ,应用CD 44V 6、MVD的单克隆抗体检测两者在 5 7例大肠腺癌、78例大肠腺瘤及 17例正常肠黏膜中的表达情况。结果　腺癌中CD 44V 6阳性表达率为 45 .6% (2 6/5 7) ,腺瘤为 5 8.0 % (3 6/62 ) ,腺瘤恶变为 75 .0 % (12 /16) ,正常肠黏膜为 5 .9% (1/17) ;腺癌中MVD值为 2 4.65± 7.84。腺癌中CD44V 6表达与MVD值有明显相关性 (P <0 .0 1) ,而且两者表达与肿瘤大小、淋巴结转移、浸润深度和分化程度密切相关。结论　CD44V 6和MVD的联合表达与大肠癌生长浸润、转移密切相关 ,联合检测CD 44V 6与MVD将有助于筛选出具有浸润复发转移的肿瘤 ,可作为判断预后及指导治疗的辅助指标之一。     

EGFR、CD44v6和C-erbB-2在结直肠癌中的表达及其与淋巴结转移、肝转移的关系

 目的 分析表皮生长因子受体（EGFR）、CD44v6和C-erbB-2在结直肠癌中的表达及其与淋巴结转移、肝转移的关系。方法 采用免疫组织化学SP法染色观察63例结直肠癌及18例结直肠腺瘤标本EGFR、CD44v6和C-erbB-2蛋白表达情况，染色结果与结直肠癌分化程度、临床分期（Duke）、淋巴结转移、肝转移等因素进行比较。结果 EGFR、CD44v6和C-erbB-2在结直肠癌与结直肠腺瘤中的阳性表达率相比较，差异均有统计学意义（P＜0.05），在结直肠癌中阳性表达率分别为68.2 %、63.5 %和47.6 %，均高于结直肠腺瘤中的阳性表达率（分别为33.3 %、11.1 %和11.1 %）。在结直肠癌伴淋巴结转移和肝转移的病例中三者联合表达较无转移者差异有统计学意义（P＜0.05）。结论 EGFR、CD44v6和C-erbB-2在结直肠癌中均有较高的阳性表达率，三者的联合表达预示着结直肠癌有较强的淋巴结转移和肝转移能力，EGFR、CD44v6和C-erbB-2可作为判定结直肠癌转移的临床生物学指标。     

黏着斑激酶的表达与结直肠癌侵袭和转移的相关性

目的 探讨黏着斑激酶(FAK)表达与结直肠癌发生及其生物学行为的关系。方法 应用免疫组织化学S—P方法，检测67例结直肠癌和配对的正常黏膜以及37例结直肠区域淋巴结转移癌组织中FAK的表达。结果 正常结直肠黏膜、结直肠癌及淋巴结转移癌组织中FAK阳性率分别为71．6％(48／67)、89．6％(60／67)和97．3％(36／37)(P=0．001)；FAK强阳性率分别为6．0％(4／67)、77．6％(52／67)和83．8％(31／37)(P=0．000)。结直肠癌中区域淋巴结有转移及区域淋巴结无转移者FAK阳性率分别为100．0％(37／37)和76．7％(23／30)(P=0．007)；FAK强阳性率分别为89．2％(33／37)和63．3％(19／30)(P=0．012)。FAK表达水平与结直肠癌浸润深度呈正相关(P=0．009)；而与结直肠癌的分化程度无关(P=0．438)。结论 FAK表达水平增高可加速结直肠恶性细胞的增殖、浸润和转移。故检测结直肠活检黏膜及癌组织中FAK表达水平，有助于进一步了解结直肠黏膜癌变倾向及结直肠癌生物学行为，有利于预后判断。     

c-myc 和 CD24在结直肠癌的表达及意义

目的：检测 c-myc 和 CD24在结直肠癌、息肉及正常黏膜中的表达,探讨 c-myc 和 CD24在结直肠癌发生中的作用以及二者之间的关系。方法采用免疫组织化学法对60例结直肠癌、45例大肠腺瘤性息肉、15例大肠增生性息肉、30例正常结直肠黏膜石蜡切片进行 c-myc 和 CD24表达的检测。结果 c-myc 阳性表达率在结直肠癌中为73.3%,明显高于大肠腺瘤性息肉44.4%（χ2=9.0168,P <0.01）、大肠增生性息肉13.3%（χ2=18.2159,P <0.01）和正常黏膜组织6.7%（χ2=35.5731,P <0.01）;CD24阳性表达率在结直肠癌中为76.7%,明显高于大肠增生性息肉6.7%（χ2=25.1330,P <0.01）、正常黏膜组织3.3%（χ2=43.1074,P <0.01）。c-myc 与结直肠癌的肿瘤部位（χ2=8.3523,P <0.01）、淋巴结转移（χ2=4.2751,P <0.05）、分化程度（χ2=4.1153,P <0.05）和分期（χ2=5.7399, P <0.05）有相关性;CD24与肿瘤大小（χ2=9.3336,P <0.01）、淋巴结转移（χ2=7.6930,P <0.01）、分化程度（χ2=5.8700,P <0.05）和分期（χ2=4.4987,P <0.05）有相关性。结直肠癌组织中 CD24与c-myc 表达呈正相关（χ2=10.8249,r =0.39,P <0.05）。结论 c-myc 与 CD24在结直肠癌中高表达,且与一些临床病理特征有关。在结直肠癌的发生、进展、转移过程中,c-myc 很可能充当了 CD24信号通路的下游靶基因,受 CD24调控。     

Villin和VEGF在结直肠腺癌组织中的表达及临床病理意义

目的 探讨绒毛蛋白(Villin)和血管内皮生长因子(VEGF)在结直肠腺癌组织中的表达及临床病理意义.方法 采用免疫组化方法检测40例结直肠腺癌、10例结直肠腺癌切缘黏膜、10例结直肠腺瘤(轻-中度不典型增生5例,重度不典型增生5例)组织中Villin和VEGF的表达情况,并对二者表达的相关性及与各临床病理特征的关系进行统计学分析.结果 Villin在结直肠腺癌切缘黏膜、结直肠腺瘤(轻-中度不典型增生、重度不典型增生)及结直肠腺癌中的阳性表达率分别为90.0％、70.0％(80.0％、60.0％)及50.0％,呈递减趋势,3组比较,差异有统计学意义(P=0.029);切缘黏膜的Villin阳性表达率高于结直肠腺癌,差异有统计学意义(P﹤0.05);Villin的表达与结直肠腺癌的病理分级、淋巴结是否发生转移以及临床病理分期相关(P﹤0.05).VEGF在切缘黏膜、结直肠腺瘤(轻-中度不典型增生、重度不典型增生)及结直肠腺癌中的阳性表达率分别为30.0％、50.0％(40.0％、60.0％)及72.5％,呈递增趋势,3组比较,差异有统计学意义(P=0.024);切缘黏膜的VEGF阳性表达率低于结直肠腺癌,差异有统计学意义(P﹤0.05);VEGF的表达与结直肠腺癌的病理分级及肌层浸润深度相关(P﹤0.05).结直肠腺癌组织中Villin和VEGF的表达不存在相关性(P﹥0.05).结论 Villin和VEGF在结直肠黏膜恶性转变中发挥一定作用,可能参与结直肠腺癌的浸润和转移.在结直肠腺癌中同时检测Villin和VEGF的表达情况,可能对判断结直肠腺癌的发生、发展及恶变程度具有一定价值,并可能有助于预测其生物学行为.     

PTTG和bFGF在结直肠癌组织中的表达

目的探讨垂体肿瘤转化基因（PTTG）和碱性成纤维细胞生长因子（bFGF）在结直肠癌组织中的表达及意义。方法应用免疫组化法检测120例结直肠癌、30例结直肠腺瘤、30例正常结直肠黏膜组织中PTTG及bFGF的表达。结果结直肠癌组织中PTTG和bFGF阳性表达率明显高于结直肠腺瘤组织及正常结直肠黏膜组织（P均〈0.05）;结直肠癌组织中PTTG和bFGF阳性表达率均与结直肠癌Dukes分期及淋巴结转移有关（P均〈0.05）。结论 PTTG和bFGF与结直肠癌生物学行为及预后有密切关系,两者联合检测有助于结直肠癌恶性程度和预后的判断。     

KLK10与TFF3在结直肠癌中的表达及与临床病理的关系

  目的  研究人组织激肽释放酶10(kallikrein 10, KLK10)和肠三叶因子(trefoil factorfamily-3, ITF/TFF3)蛋白在结直肠腺癌、腺瘤和正常结直肠黏膜组织中的表达情况及相互作用关系, 探讨其在结直肠癌发生发展中的作用及与临床、病理的关系。  方法  采用SP免疫组织化学染色法检测57例结直肠腺癌、21例结直肠腺瘤及11例正常结直肠黏膜组织KLK10和TFF3的表达情况, 并分析KLK10和TFF3的表达与结直肠癌临床病理指标的关系。  结果  KLK10和TFF3在结直肠腺癌的阳性率分别为66.7%(38/57)和70.2%(40/57), 在结直肠腺瘤的阳性率分别为38.1%(8/21)和47.6%(10/21), 在正常结直肠黏膜组织中的阳性率为27.3%(3/11)和90.9%(10/11)。KLK10和TFF3在结直肠腺癌、结直肠腺瘤及正常结直肠黏膜组织中的表达均具有显著性差异(P均 < 0.0.05)。KLK10和TFF3的表达均与患者的TNM分期及淋巴结转移情况相关(P均 < 0.0.05)。KLK10和TFF3在结直肠癌中的表达呈正相关(r=0.597, P < 0.001)。TFF3在KLK10阳性组中的表达明显高于在KLK10阴性组中的表达。  结论  KLK10和TFF3参与结直肠癌的发生发展并与结直肠癌的TNM分期及淋巴结转移相关, 可作为较为理想的肿瘤标志物辅助判断患者预后并指导临床治疗。      

Prognostic and diagnostic significance of beta-catenin nuclear immunostaining in colorectal cancer.

In the present study, we investigated the prognostic and diagnostic significance of beta-catenin nuclear immunostaining in 60 specimens of normal colorectal tissue; 180 specimens of colorectal polyps, adenomas, and carcinomas; and 40 specimens from patients with the simultaneous occurrence of polyps, adenomas, and carcinomas. Additional specimens from 59 patients with colorectal carcinoma and 14 patients with adenoma who subsequently developed carcinoma were examined for possible survival study. Immunohistochemical staining showed that the occurrence of nuclear beta-catenin correlated with the sequential stages in colorectal carcinogenesis, in which positive staining was observed in 0% of normal tissues, 8% of polyps, 92% of adenomas, and 100% of carcinomas. High immunohistochemical scores in colorectal carcinoma were significantly associated with lymph node metastasis and poor survival. Adenomas associated with synchronous or metachronous carcinomas showed significantly higher levels of nuclear beta-catenin compared with adenomas without associated carcinomas. Nuclear translocation of beta-catenin was rare or absent in other types of cytokeratin 20 positive adenocarcinomas examined (99 cases). Thus, it was positive in only 7% of colonic mucinous adenocarcinomas, 3% of pancreatic adenocarcinomas, 8% of ovarian mucinous cystadenocarcinomas, and 0% of gastric adenocarcinomas. However, 100% of primary and metastatic colorectal adenocarcinomas were positive for nuclear staining for beta-catenin. Thus, nuclear staining for beta-catenin may serve as an additional parameter to help distinguish colorectal adenocarcinomas from adenocarcinomas of other tissue sites. Collectively, the present large-scale study has clearly addressed the clinical significance of beta-catenin nuclear translocation with respect to tumor progression, survival, and differential diagnosis.     

大肠肿瘤中bcl—2及PCNA表达及其意义

目的：研究ｂｃｌ－２及ＰＣＮＡ在大肠肿瘤肆生中的作用及其临床意义。方法：应用免疫组化Ｓ－Ｐ法分别检测大肠正常粘膜、腺瘤及腺癌中ｂｃ；－２及ＰＣＮＡ表达。结果：Ｂｃｌ－２在正常粘膜基底部上皮细胞表达，在腺瘤（７７．５％）和腺癌中（５６．３％）ｂｃｌ－２表达差异显（Ｐ〈０．０５）。高分化腺癌ｂｃｌ－２表达率（６８．４％）显高于差分化腺癌（４１．７％），ＰＣＮＡ表达在正常粘膜，腺瘤及腺癌中依次递增，     

Interleukin-4 receptor and epidermal growth factor receptor expression in colorectal cancer.

Interleukin-4 receptor (IL-4R) and Epidermal Growth Factor receptor (EGFR) were assessed as factors associated with adenoma-carcinoma progression in colorectal cancer and tumour invasion. A monoclonal antibody (MR6) was applied to detect IL-4R in: metaplastic polyps (five cases), adenomas (15 cases), and carcinomas (44 adenocarcinomas and one squamous cell). Positive labelling was obtained in all polyps, adenomas and in 40/45 carcinomas. Normal colonic mucosa of these patients, as well as macrophages and lymphocytes infiltrating the tumour stroma, were also positively labelled with MR6. Four out of five poorly differentiated adenocarcinomas did not show IL-4 receptor expression. No significant correlation was found with tumour size, lymph node stage and IL-4 receptor expression. On the above specimens a parallel detection of epidermal growth factor receptors (EGFR) by a monoclonal antibody (EGFR 1) was carried out. Expression of EGFR was found in 14/20 polyps and in 22/45 carcinomas. All but one of the EGFR positive malignant tumours showed coexpression of IL-4 receptor. Lymph node involvement by tumour cells was detected in 25 out of 45 patients. Eighteen of these 25 cases were positive with EGFR1.     

c-MET expression in colorectal adenomas and primary carcinomas with its corresponding metastases

Background

c-MET plays an important role in tumor proliferation, invasion and metastasis. In this study we examined the expression of c-MET in colorectal adenomas, primary adenocarcinomas and their corresponding lymph node, peritoneal and liver metastases. We correlated our findings with clinicopathological features.

Methods

Twenty three cases of colorectal adenoma and 102 cases of primary colorectal carcinoma and their corresponding metastases (44 lymph nodes, 21 peritoneal deposits and 16 liver metastases) were studied to evaluate c-MET expression by immunohistochemistry. For comparison, 12 sections of adjacent healthy colorectal mucosa were examined.

Results

Statistically significant differences were present among normal tissues, colorectal adenomas and primary colorectal carcinomas (P=0.011). Normal tissues showed a negative or weak reaction in 66.67% and 33.33% of cases respectively. Expression of c-MET was positive in 47.8% of adenomas. A significant positive association was identified between c-MET high expression and degree of dysplasia (P=0.024). c-MET was highly expressed in 66.7% of primary colorectal carcinoma. Significant positive correlations were detected between c-MET expression and TNM stage (P=0.036), lymph node metastasis (LNM), peritoneal deposits and liver metastasis (P=0.038, P=0.094 and P=0.045, respectively). c-MET expression in metastatic tissues was significantly higher than that of the primary tumor.

Conclusions

c-MET expression is gradually up-regulated in the development and progression of colorectal cancer (CRC) from normal epithelium to adenoma to colorectal carcinoma to metastases.     

MMP—2、VEGF、CD44V6、E—cd在大肠癌组织中的表达

目的 探讨基质金属蛋白酶－2（MMP－2）、血管内皮生长因子（VEGF）、CD44拼接异构体（CD44V6）、上皮钙粘附蛋白（E－cd）在大肠癌组织中的表达及其与转移的关系。方法 采用免疫组化SLAB法对45例大肠癌组织进行免疫组化研究;取10例大肠腺瘤样息肉和8例正常大肠粘膜细胞作对照。结果 大肠癌DukesA、B期和C、D期MMP－2、VEGF、CD44V6、E－cd的阳性表达率分别为18.2%（4／22）、63.6%（14／22）、27.3%(6/22)、100.0％（22／22）和91.3%（21／23）、78.3%（18／23）、87.0%（20／23）、69.6%（16／23）;对照组MMP－2、VEGF、CD44V6、E－cd的阳性表达率为5.6%(1/18)、33.3%（6／18）、5.6(1/18)、100.0%(18/18)。Dukes C、D期和A、B期的MMP－2、CD44V6和E－cd的表达有非常显著性差异（P＜0.01）,而VEGF的表达无显著性差异（P＞0.05）;大肠癌与对照组间MMP－2、CD44V6和VEGF的表达有非常显著性差异（P＜0.01）,而DukesC、D期和对照组间E－cd的表达有非常显著性差异（P＜0.01）。结论 MMP－2、CC44V6与大肠癌的分期和转移密切相关;E－cd、VEGF与转移无明显关系。     

Expression of p-glycoprotein in the normal colorectal epithelium and in colorectal-carcinoma

Expression of P-glycoprotein (pgp) was analyzed by immunohistochemical staining with monoclonal antibody JSB-1 in 145 frozen specimens (67 were samples of normal colorectal mucosa from sites adjacent to the tumor, 66 were colorectal carcinomas, 5 colorectal polyps, 5 metastatic lymph nodes, and 2 samples of metastatic liver tumors) of 67 patients with colorectal carcinoma and polyps. All 72 specimens of normal colorectal mucosa and adenomatous polyps expressed pgp to various degrees. By contrast, 18 of 39 (46.2%) samples from cases of well differentiated adenocarcinoma were positive for pgp but only 3 of 21 (14.3%) samples from cases of moderately differentiated adenocarcinoma and none of the 4 samples from cases of poorly differentiated adenocarcinoma were positive for pgp. There was no correlation between the clinicopathological stage of colorectal carcinoma and the expression of pgp. These findings indicate that the expression of P-glycoprotein is closely related to the differentiation of cells. In normal colorectal epithelium, pgp was expressed normally and in well differentiated adenocarcinomas, pgp was still expressed. However, expression of pgp was no longer detectable in carcinomas with moderate or poor differentiation.     

CD44粘附分子表达与肿瘤转移相关性的探讨

目的 探讨CD44粘附分子表达与肿瘤转移相关性的意义.方法 采用LSAB免疫组化法对淋巴结转移性腺癌31例,鳞癌27例,和伴有淋巴结转移的鼻咽癌20例,食管鳞癌39例的原发癌石蜡切片进行CD44单抗染色,观察其表达阳性率及反应强度.结果1.淋巴结转移性腺癌CD44阳性表达18例(57%),转移性鳞癌阳性表达7例(2%),两组间腺癌阳性表达率明显高于鳞癌(P0.05).2.伴有淋巴结转移的鼻咽癌阳性表达11例,占55%(11/20),食管鳞癌阳性表达25例,(64%).两组间阳性表达率无明显差异(P>0.05),且阳性反应强度亦无显著的差异(P>0.05).结论 在多数肿瘤CD44表达与肿瘤转移密切相关,而以CD44V为明显,但无一致性.因此CD44不能作为肿瘤转移通用性标记物.     

Abnormalities in the expression of cell cycle-related proteins in tumors of the small bowel.

Tumors of the small bowel are quite rare for unknown reasons, although they resemble colorectal tumors in many respects. The purpose of this study was to determine whether abnormalities in the expression of several cell cycle control genes are of importance in small bowel tumorigenesis by comparing a series of samples of normal mucosa, adenomatous polyps, and adenocarcinomas. The levels of cyclin D1, cyclin E, p16, p21, p27, and p53 proteins were determined by immunohistochemistry in samples of normal small bowel (n = 16), small bowel adenomas (n = 20), and small bowel adenocarcinomas (n = 24). Normal small bowel mucosa expressed p27 protein, but not the other cell cycle-related proteins. About 20% of the tumors displayed a decrease in the expression of this protein. The most frequent alteration in the tumors was an increase in the p16 protein. Increased expression of p53 was associated with tumor progression because it was overexpressed in 45% of the adenomas and 65% of the adenocarcinomas (P<0.05). Advanced age and increased detection of cyclin D1 and p53 were associated with a decreased 3-year survival (P<0.05). Cell cycle abnormalities are early and important events in the multistep process of small bowel tumorigenesis, thus resembling colorectal carcinogenesis. As in colon cancer, deregulated expression of G1 proteins may perturb cell cycle control in benign adenomas of the small bowel and thereby enhance tumor progression. Increased expression of cell cycle inhibitors in tumors may serve as a defense mechanism for tumor progression.