热休克蛋白10在大肠癌的表达及其临床病理意义

目的研究热休克蛋白10(heat shock protein 10,HSP10)在大肠癌发生、发展过程中的表达规律,了解其与大肠癌临床病理特征和术后生存期的关系。方法应用免疫组化EnVision二步法,检测HSP10在大肠正常黏膜、腺瘤和腺癌中的表达,及其与临床病理表现和术后生存期的相关性。结果HSP10在大肠正常黏膜、腺瘤和腺癌中均有表达,在腺瘤、腺癌中的表达明显强于正常黏膜(P<0.001);HSP10的阳性表达与大肠癌患者的年龄、性别、肿瘤浸润深度、局部淋巴结转移、远处转移、临床分期、组织学分化程度等临床病理特征和术后生存期均无关。结论HSP10在大肠腺瘤和腺癌的高表达,提示可能与大肠癌的发生发展有关;HSP10的阳性表达与大肠癌的临床病理特征和术后生存期无关。     

大肠腺瘤和腺癌组织中Survivin的表达及其意义

目的:研究Survivin在大肠腺瘤和腺癌组织中的表达及其意义。方法:SP法检测大肠腺瘤50例(管状腺瘤35例,绒毛状腺廇9例,混合腺廇6例)和大肠腺癌89例组织中Survivin表达情况,正常大肠黏膜组织20例作为对照。结果:在正常大肠黏膜组织、腺瘤和大肠腺癌中,Survivin的阳性表达率分别为10.0%(2/20)、26.0%(13/50)和75.3%(67/89);随腺瘤不典型增生程度增高,Survivin的表达也随之明显增高,P=0.019。Survivin的表达随大肠腺瘤分化程度的降低而升高[高、中和低分化组阳性表达率分别为65.0%(26/40)、86.0%(37/43)和100%(6/6)]。伴有淋巴结转移组大肠腺癌Survivin阳性表达率85.2%(52/61)明显高于无淋巴结转移组53.6%(15/28),P=0.003。在生存时间≥2年的大肠腺癌中,阳性表达率为73.7%(14/19),在<2年组中为64.7%(11/17),两者差异无统计学意义,P=0.41。结论:正常大肠黏膜、大肠腺瘤和大肠腺癌中Survivin的阳性表达逐步增高。随大肠腺瘤不典型增生程度增高,Survivin的表达也随之明显增高。Survivin的表达随大肠腺癌分化程度的降低而升高。伴有淋巴结转移组大肠腺癌Survivin表达的阳性率明显高于与无淋巴结转移组。     

核转录因子E2F-1和p27在大肠癌中的表达及意义

目的 探讨E2F-1和p27在大肠腺癌组织中的表达及意义.方法 采用免疫组织化学S-P法检测78例大肠癌、20例正常大肠黏膜和30例大肠腺瘤E2F-1和p27的表达情况.结果 E2F-1在大肠癌中的阳性率显著高于正常大肠黏膜和大肠腺瘤(P＜0.05).E2F-1表达与大肠癌的病理分化程度、淋巴结转移和临床分期具有相关性(P＜0.05).p27在正常大肠黏膜的阳性率明显高于大肠腺瘤和大肠癌.p27表达与大肠癌的病理分化程度、淋巴结转移和临床分期具有相关性(P＜0.05).结论 E2F-1和p27的表达与大肠癌发生关系密切,可以作为肠癌发生、发展的生物学标志物.     

c-erbB-2、ras、p53基因产物在大肠癌及癌前病变中的表达

目的：探讨c－erbB－2、ras、p53基因的表达与大肠癌发生发展的关系以及其对大肠癌早期诊断、预后判断的价值。方法：对45例大肠癌及36例癌旁非腺瘤型不典型增生、17例大肠腺瘤，用免疫组化方法检测基因产物的表达。结果：p53在大肠癌的阳性表达率为57．8％，p53蛋白高表达的癌旁非腺瘤型不典型增生及大肠腺瘤均为中度或高度不典型增生。p53蛋白表达与大肠癌的组织分化程度、淋巴结转移有关（P＜0．05）。p21、p185蛋白表达与大肠癌组织学分型、癌组织浸润程度、淋巴结转移无关，而与癌组织分化程度有关（P＜0．05）。结论：p53蛋白表达可能是大肠病变恶性倾向的一个独立标志。p53、p21、p185蛋白对大肠癌的发生、发展起重要作用。     

COX-2和VEGF-C在大肠癌中的表达及其临床意义

目的探讨环氧合酶（COX-2）和血管内皮生长因子-C（VEGF-C）在大肠癌中的阳性表达率与临床病理特点,特别是与淋巴结转移的关系。方法选择经病理确诊的大肠癌60例,同体癌组织远端5 cm以外的正常大肠组织30例和大肠腺瘤30例为研究对象。采用免疫组化S-P染色法,检测三组的COX-2和VEGF-C的表达率,进行比较及关联性分析,并将大肠癌的阳性表达率与淋巴结转移等临床病理特点进行相关性分析。结果 （1）COX-2正常大肠组织不表达;大肠癌和大肠腺瘤的阳性表达率均高于正常大肠组织（P〈0.017）;COX-2大肠癌的阳性表达率,在中低分化、浸及浆膜、有淋巴结转移和C＋D期均比相应的高分化、未及浆膜、无淋巴结转移和A＋B期增高（P〈0.05,OR〉1,其95%可信区间内不包含1）。（2）VEGF-C在正常大肠组织和大肠腺瘤均不表达;大肠癌的阳性表达率均高于大肠腺瘤和正常大肠组织（P〈0.017）;VEGF-C阳性表达率,在中低度分化、有淋巴结转移和C＋D期均比相应的高分化、无淋巴结转移和A＋B期高（P〈0.01,OR〉1,其95%可信区间内不包含1）。（3）大肠癌中,COX-2和VEGF-C的表达有关联性,即COX-2表达高,VEGF-C亦高（P〈0.05,OR〉1,其95%可信区间内不包含1）。结论在大肠癌中,COX-2和VEGF-C均呈高表达,且有关联性;阳性表达率与淋巴结转移、Dukes分期和组织分化有相关性。     

大肠癌组织COX-2的表达及其与肿瘤细胞增生和凋亡的关系

目的 探讨大肠癌中环氧化酶-2（c yclooxygenase-2,COX-2）表达及其与癌细胞增生和凋亡的关系.方法 采用免疫组织化学法检测60例大肠癌、20例大肠腺瘤和20例癌旁正常大肠黏膜中COX-2和PCNA的表达,并采用TUNEL法检测原位细胞凋亡水平.结果 大肠癌组织中COX-2阳性表达、PI及AI均显著高于大肠腺瘤及正常大肠黏膜组织（P＜0.05）;COX-2的高表达与大肠癌肿瘤大小、淋巴结转移和临床分期有关,与肿瘤生长部位、分化程度无关.COX-2阳性大肠癌组中PI高于COX-2阴性大肠癌组,AI低于COX-2阴性大肠癌组（P＜0.05）.结论 COX-2的过度表达可促进大肠癌细胞增生、抑制细胞凋亡,在大肠癌的发生、发展过程中起着重要的作用.     

APC、β-catenin及其意义和c-myc在大肠癌中的表达

摘要：目的探讨APC、β-catenin和c-myc在大肠癌发生、发展中的作用。方法采用免疫组化方法检测30例正常大肠黏膜、30例大肠腺瘤、10例大肠腺瘤恶变及50例大肠癌组织中APC、pcatenin和c-myc蛋白的表达情况。结果大肠癌和大肠腺瘤恶变APC阳性率分别为44．0％、40．0％，显著低于大肠腺瘤（86．7％）和正常大肠黏膜（100％）（P〈0.01）。大肠癌、大肠腺瘤恶变和大肠腺瘤β-catenin胞浆和（或）胞核异位表达率分别为62．0％、50．0％、30．0％，显著高于正常大肠黏膜（0）（P〈0.01），大肠癌8Icatenin异位表达率显著高于大肠腺瘤（P〈0.01）。大肠癌、大肠腺瘤恶变、大肠腺瘤c-myc阳性率分别为56．0％、60．0％、46．7％，显著高于正常大肠黏膜（0）（P〈0.01）。大肠癌中8-catenin膜表达缺失率为：46．0％，显著高于大肠腺瘤（10．0％）和正常大肠黏膜（0）（P〈0.01），且与大肠癌组织分化程度、浸润深度、淋巴结转移、Dukes分期有关。APC蛋白表达与大肠癌组织分化程度有关。大肠癌中8Icatenin异位表达与c-myc阳性表达呈正相关关系（r=0．63，P〈0.01），而与APC阳性表达呈负相关关系（r=一0．39，P％0．05）。结论APC失表达和（或）pcatenin异位表达，激活癌基因c-myc过表达与大肠癌的发生、发展密切相关，可能是大肠癌发生的早期事件；pcatenin膜表达缺失与大肠癌的侵袭、转移有关。     

APC、β-catenin 和c-myc 在大肠癌中的表达及其意义

 目的 探讨APC、β-catenin和c-myc在大肠癌发生、发展中的作用。方法 采用免疫组化方法检测30例正常大肠黏膜、30例大肠腺瘤、10例大肠腺瘤恶变及50例大肠癌组织中APC、pcatenin和c-myc蛋白的表达情况。结果 大肠癌和大肠腺瘤恶变APC阳性率分别为44．0％、40．0％，显著低于大肠腺瘤（86．7％）和正常大肠黏膜（100％）（P〈0.01）。大肠癌、大肠腺瘤恶变和大肠腺瘤β-catenin胞浆和（或）胞核异位表达率分别为62．0％、50．0％、30．0％，显著高于正常大肠黏膜（0）（P〈0.01），大肠癌8Icatenin异位表达率显著高于大肠腺瘤（P〈0.01）。大肠癌、大肠腺瘤恶变、大肠腺瘤c-myc阳性率分别为56．0％、60．0％、46．7％，显著高于正常大肠黏膜（0）（P〈0.01）。大肠癌中8-catenin膜表达缺失率为：46．0％，显著高于大肠腺瘤（10．0％）和正常大肠黏膜（0）（P〈0.01），且与大肠癌组织分化程度、浸润深度、淋巴结转移、Dukes分期有关。APC蛋白表达与大肠癌组织分化程度有关。大肠癌中8Icatenin异位表达与c-myc阳性表达呈正相关关系（r=0．63，P〈0.01），而与APC阳性表达呈负相关关系（r=一0．39，P％0．05）。结论 APC失表达和（或）pcatenin异位表达，激活癌基因c-myc过表达与大肠癌的发生、发展密切相关，可能是大肠癌发生的早期事件；pcatenin膜表达缺失与大肠癌的侵袭、转移有关。     

抑癌基因PTEN在大肠癌癌变过程中表达的实验研究

目的探讨抑癌基因PTEN的表达与大肠癌发生及浸润转移的关系。方法应用免疫组织化学EnVi-sion法原位观察正常大肠粘膜、癌旁单纯增生上皮、增生性息肉、腺瘤型息肉及癌组织中PTEN蛋白的表达。结果正常大肠粘膜均见PTEN蛋白的表达。癌旁单纯增生粘膜、增生性息肉、腺瘤型息肉及癌组织中PTEN蛋白的阳性表达率分别为92.3%、87.5%、70.6%及56.9%,其中早期癌的阳性表达率为66.7%,浸润癌的阳性表达率为55.9%。无淋巴结转移者的阳性表达率为57.5%,有淋巴结转移者为32.0%(P=0.0002)。PTEN蛋白阳性表达率随肿瘤分化程度降低而降低(P=0.028),随肿瘤Dukes分期的增加而下降,但无统计学意义(P>0.05)。结论PTEN蛋白的表达与大肠癌的发生、浸润及淋巴结转移有关,其低表达可能是大肠癌发生的一个信号。     

PCNA和EGFR在不同性质大肠肿瘤中表达的研究

目的探讨PCNA和EGFR在不同性质大肠肿瘤中的表达及意义。方法采取正常大肠粘膜10例,大肠腺瘤20例,大肠癌粘膜30例。应用免疫组化染色方法（SP法）,在连续切片上同步检测PCNA和EGFR的表达。结果PCNA染色阳性物质主要定位于细胞核,EGFR染色阳性物质主要定位于细胞膜。PCNA和EGFR阳性细胞在大肠腺瘤和大肠癌中的分布呈不同程度的异质性。正常大肠粘膜、大肠腺瘤和大肠癌粘膜PCNA阳性细胞数分别为2．42±1．02、10．17±4．96和20．21±6．58,各组间比较均有极显著性差异（P＜0．01）;EGFR阳性率分别为0、30％和73．33％．各组间比较也有显著性差异（P＜0．05）。PCNA和EGFR的表达与大肠癌的分化程度、分期和淋巴结转移有关,与组织学类型无关。30例大肠癌粘膜PCNA和EGFR的表达里显著性正相关（P＜0．05）;60例不同性质大肠粘膜PCNA和EGFR的表达里极显著性正相关关系（P＜0．01）。结论PCNA和EGFR都是反映细胞增殖活性的重要指标。结合病理形态学观察,同时检测PCNA和EGFR的表达,能更客观更准确地判断大肠癌的恶性程度和生物学特性。     

热休克蛋白10在大肠癌的表达及其临床病理意义

Objective： To investigate the expression of heat shock protein 10 （HSPIO） during genesis and development of large bowel carcinoma and discuss the clinical significance about its expression. Methods： The expression of HSPIO was observed in specimens from normal colonic mucosa （NC）, colorectal adenomas （CA） and colorectal adenocarcinomas （CAC） by immunohistochemistry EnVisionTM. Its correlations to clinicopathologic features, as well as to postoperative survival time of large bowel carcinoma patients were analyzed. Results： The expression of HSPIO was common in normal colonic mucosa, colorectal adenomas and adenocarcinomas and more intensive in colorectal adenomas and adenocarcinomas than that in normal colonic mucosa （P 〈 0.001）. The positive expression of HSPIO had no correlation to clinicopathologic features, including age, gender, primary tumor, infiltrating of regional lymph node, metastasis, clinical stage and histopathology of large bowel carcinoma patients, as well as to their postoperative survival time. Conclusion： HSPIO was overexpressed in the early stage of colorectal adenocarcinoma suggesting that it could serve as an index for early diagnosis of large bowl carcinoma. The positive expression of HSPIO had no correlation to clinicopathologic features or postoperative survival time of large bowel carcinoma patients.     

Matrix Metalloproteinase-9 Expression in the Normal Mucosa–Adenoma–Dysplasia–Adenocarcinoma Sequence of the Colon

It has been proposed that matrix metalloproteinases (MMPs) play a role in tumor invasion. We determined protein expression of matrix metalloproteinase-9 (MMP-9) in colorectal cancer (CRC), corresponding normal mucosa and colorectal adenomas. For confirmation of immunohistochemical results MMP-9 TaqMan RT-PCR analysis was performed. Expression of MMP-9 was determined on paraffin embedded biopsy sections by immunohistochemistry in 31 CRC patients (from cancer tissue and corresponding normal mucosa) and in 30 patients with adenoma (nine adenomas with high grade of dysplasia). MMP-9 immunostaining was determined semi-quantitatively. For Taqman RT-PCR analyses normal mucosa (n = 5), adenoma without (n = 6) and with high grade dysplasia (n = 7) and CRC (n = 10) were investigated. Statistical analysis with ANOVA, LSD test and correlation analysis were performed. P value of <0.05 was considered significant. The MMP-9 expression in CRC was significantly higher compared to adenomas or the normal mucosa (P = 0.001). Significantly higher expression of MMP-9 has been observed in adenomas with high grade dysplasia compared to other adenomas or normal colon (P < 0.001). Diffuse strong MMP-9 expression was present in tumor as well as in stromal cells. In adenoma samples, dysplastic epithelial cells showed moderate intensive cytoplasmic MMP-9 expression, with a clear-cut differentiation between dysplastic and non-dysplastic areas. Staining intensity correlated with the grade of CRC. We demonstrate a significantly higher expression of MMP-9 in adenoma with high grade dysplasia-CRC sequence as compared to normal tissue. The over-expression of MMP-9 strongly suggests its association with colorectal carcinogenesis.     

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，     

CD44V6在结直肠癌组织中的表达及临床意义

目的：探讨CD44V6表达与结直肠癌发生、发展及转移的关系。方法：采用S－P免疫组织化学方法，检测78例原发性结直肠腺癌、14例结直肠腺瘤癌变、57例结直肠腺瘤、30例结直肠增生性息肉和24例结直肠正常黏膜中CD44V6的表达情况。结果：腺瘤、腺瘤癌变和腺癌组织中CD44V6阳性表达率分别为78．95％、100．00％和56．41％，明显高于增生性息肉和正常黏膜组织的阳性表达率（14．81％）。CD44V6阳性表达与腺癌淋巴结转移、Dukes分期和病理分级无相关性。结论：CD44V6表达与结直肠癌的发生有关，可作为诊断结直肠癌前病变和早期癌的生物学指标。     

Cyclin B1, unlike cyclin G1, increases significantly during colorectal carcinogenesis and during later metastasis to lymph nodes

In collaboration with p53, cyclins B1 and G1 regulate the G2/M transition, a key checkpoint in the active cell cycle, which can be monitored by Ki67. However, the cyclin B1 expression remains unclear during colorectal carcinogenesis and during later metastasis to lymph nodes, and cyclin G1 expression is not clear in colorectal tumors. To clarify the variations of the two cyclins in colorectal tumors, cyclin B1, cyclin G1, p53, and Ki67 were immunohistochemically stained in 22 normal mucosa, 62 adenomas, 17 carcinomas in adenomas, 194 primary carcinomas, and 21 lymph node metastases; and the two cyclins were examined by Western blot in other 10 pairs of normal mucosa and primary carcinomas. Located in cytoplasms, nuclei or both, cyclin B1 expression increased significantly from normal mucosa through adenomas to primary carcinomas, from adenomas with mild dysplasia through those with moderate to those with severe, from peripheral adenomas to their central carcinomas, and from primary to metastatic foci. These increased expressions were confirmed by Western blot. Cyclin B1 expression, however, declined significantly in primary carcinomas showing large size, mucinous type, deep invasion, or short postoperative-patient-survival time. High cyclin B1 was linked to high p53 in adenomas, and to high Ki67 in adenomas and primary carcinomas. In contrast, found limited to nuclei, cyclin G1 expression did not vary significantly from normal mucosa through to metastatic carcinomas, and was not associated with clinicopathological parameters, p53 or Ki67. The unchanged expressions were confirmed by Western blot. Thus, increased cyclin B1, but not cyclin G1, may promote colorectal carcinogenesis and later metastasis to lymph nodes.     

Loss of p57KIP2 is associated with colorectal carcinogenesis

The expression and significance of p57KIP2, an important inhibitor of the cell cycle, remain unclear during carcinogenesis and during late metastasis to lymph nodes of tumors. To detail changes of p57KIP2 during colorectal carcinogenesis and during late metastasis to lymph nodes, p57KIP2, cyclin A, cyclin B1, cyclin E, CDK2, and Ki67 were immunohistochemically investigated in 22 specimens of normal mucosa, 62 of adenomas, 17 of carcinomas in adenomas, 189 of primary carcinomas, and 23 of lymph node metastases. Situated in nuclei, p57KIP2 expression increased significantly from normal mucosa to adenomas (p=0.0068), from mild through moderate to severe dysplasia in adenomas (p=0.0132). It significantly decreased from adenomas to unpaired primary carcinomas (p=0.0112) and from peripheral adenomas to paired central carcinomas (p=0.0018), but remained unchanged when primary carcinomas metastasized to lymph nodes (p=0.3401). p57KIP2 expression was not correlated with clinicopathological indices, but the patients having tumors without p57KIP2 tended to show a poor prognosis (p=0.0674). High p57KIP2 was significantly correlated with increased cyclin A (p=0.0007), elevated cyclin B1 (p=0.0007), reduced CDK2 (p=0.0021), and increased Ki67 (p=0.0013) in adenomas. Thus, loss of p57KIP2 expression appears associated with colorectal carcinogenesis.     

Correlation of Skp2 with carcinogenesis, invasion, metastasis, and prognosis in colorectal tumors

In colorectal tumors, S-phase kinase-associated protein 2 (Skp2) still has numerous important questions unanswered: its expression in adenomas, its correlation with key clinicopathological indices, its association with patient prognosis, its variation in lymph node metastases, and its association with many cell-cycle regulators. To answer these questions in colorectal tumors, Skp2, cyclin A, cyclin B1, cyclin E, CDK2, and Ki67 were immunohistochemically stained in 12 normal mucosa, 36 adenomas, 11 carcinomas in adenomas, 102 primary carcinomas, and 12 paired lymph node metastases; and Skp2 was examined by Western blot in 8 pairs of normal mucosa and carcinomas. Situated in nuclei, Skp2 expression significantly increased from normal mucosa through adenoma to primary carcinoma (p<0.0001), from mild through moderate to severe dysplasia in adenomas (p=0.038), from peripheral adenoma to paired central carcinoma (p=0.0033), and from primary carcinoma to lymph node metastasis (p=0.015), and these increases were confirmed by Western blot. Expression, however, relatively declined significantly in the primary carcinomas showing deep invasion (p=0.0113), lymph nodal metastases (p=0.0268), and poor prognosis for all (p=0.0104) or stage III patients (p=0.0119). High Skp2 was also significantly linked with elevated cyclin A, cyclin B1, cyclin E, CDK2 (in primary carcinomas only), and Ki67 in both adenomas and primary carcinomas. Thus, overexpression of Skp2 is associated with colorectal carcinogenesis and late metastasis to lymph nodes, whereas relative reduction of Skp2 is correlated with local invasion of primary carcinoma.     

Detection of the tumor-associated glycoprotein antigen (TAG-72) in premalignant lesions of the colon

We used monoclonal antibody B72.3 to study the expression of the colorectal carcinoma-associated antigen TAG-72 in premalignant colonic lesions with the immunoperoxidase technique. This antigen, which is rarely detectable in the normal colonic epithelium, was expressed in 13 of 19 adenomas with moderate to severe dysplasia and nine of nine cases of inflammatory bowel disease. The antibody reacted with the normal-appearing mucosa adjacent to a carcinoma in 10 of 12 cases, although only eight of the tumors expressed the antigen. The expression of the TAG-72 antigen in the colonic epithelium may be an early marker of malignant transformation.     

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.