乙型肝炎病毒相关肝细胞癌P^53蛋白的免疫组织化学研究

采用免疫组化染色检测肝癌组织及癌旁肝组织ＨＢｓＡｇ，ＨＢｃＡｇ以及Ｐ＾５３蛋白的表达，结果显示：癌灶组织中Ｐ＾５３蛋白阳性率２６．７％（１２／４５），癌旁肝组织无１例Ｐ＾５３蛋白阳性，Ｐ＾５３蛋白表达与ＨＢｓＡｇ无明显关系，而与肝癌发生年龄及癌细胞分化程度有关，提示Ｐ＾５３基因突变与ＨＢＶ相关肝癌的发生有一定关系，而多数肝癌组织无并Ｐ＾５３基因突变又进一步提示肝癌发生尚涉及其它复杂机制，此外Ｐ６５     

直肠癌内分泌样癌细胞分化与P^53蛋白表达关系的研究

目的：研究直肠癌内分泌样癌细胞分化与Ｐ＾５３蛋白表达的关系及意义。方法：采用免疫组织化学ＡＢＣ法检测４４例福尔马林固定、石蜡包埋的直肠癌手术切除及活检组织中的嗜铬蛋白Ａ（ＣＧＡ）及Ｐ＾５３蛋白的表达。结果：Ｐ＾５３、ＣＧＡ阳性表达率分别为６８．２％和３８．６％：ＣＧＡ阳性及阴性直肠癌Ｐ＾５３表达率分别为８８．２％和５５．６％，两者比较Ｐ〈０．０２５Ｘ＾２检测；Ｐ＾５３ＣＧＡ在分化差的直肠癌中的表达     

肝癌P^53基因突变与HBxAg关系的免疫组化研究

本实验对４８例石蜡包埋人肝癌和癌旁组织的Ｐ＾５３蛋白（突变型）和ＨＢｘＡｇ进行了免疫组化（ＡＢＣ法）检测。结果显示肝癌及其癌旁组织Ｐ＾５３蛋白（突变型）和ＨＢｘＡｇ阳性率分别为２９．２％（１４／２８）和９３．８％（４５／１８），Ｐ＾５３蛋白（突变型）与ＨＢｘＡｇ表达不相关（Ｐ〉０．０５），提示ＨＢｘＡｇ在Ｐ＾５３基因突变中的不发挥作用。     

胃部组织P^53蛋白免疫组化研究及临床病理学特征和预后的关系

利用免疫组化ＬＳＡＢ法，检测３６例胃癌患者胃癌组织石蜡切片标本。结果发现Ｐ＾５３蛋白在胃癌组织中阳性率为４４．４％。晚期胃癌Ｐ＾５３蛋白阳性率显著高于早期胃部；Ｐ＾５３蛋白阴性者生存时间长于Ｐ＾５３蛋白阳性者。提示Ｐ＾５３蛋白表达与胃癌患者预后有在。     

血清HBsAg阳性患者肝癌及癌旁组织中P^53和K—ras基因突变及其相…

应用ＰＣＲ－ＳＳＣＰ方法对２４例血清ＨＢｓＡｇ患者肝癌及癌旁组织中Ｐ＾５３和Ｋ－ｒａｓ基因突变进行了检测。肝癌组织中Ｐ＾５３基因突变率为３３％，Ｋ－ｒａｓ基因突变率为２５％，癌旁正常组织中无Ｐ＾５３和Ｋ－ｒａｓ基因突变，癌旁硬化组织中Ｋ－ｒａｓ突变为１５％，无Ｐ＾５３基因突变。     

P^16蛋白在子宫内膜癌中的表达

应用免疫组织化学技术检测１０例正常子宫内膜、２２例不典型增生和４２例子宫内膜癌组织Ｐ＾１６蛋白的阳性表达率。结果显示,子人膜癌组织Ｐ＾１６蛋白阳性表达率为６９％,低于正常子宫内膜（１００％）和不典型增生组织（９１％）,高分化肿瘤的Ｐ＾１６蛋白阳性表达率高于低分化者,但Ｐ＾１６蛋白的阳性表达率与子宫内膜癌临床分期无关;子宫内膜癌患者Ｐ＾１６帽白表达阴性者较阳性者预后差。提示Ｐ＾１６基因可能参与子宫内     

P^53及nm23基因异常表达与肝细胞肝癌临床,病理的关系

用免疫级化方法检测了２７例肝细胞肝癌患者癌组织Ｐ＾５３及ｎｍ２３基因异常表达，结果Ｐ＾５３及ｎｍ２３基因异常表达率分别为６６．６７％和５９．２６％。Ｐ＾５３及ｎｍ２３基因异常表达与患者血清ＡＦＰ阳性有关（Ｐ〉０．０５），与存在门静脉癌栓有关（Ｐ〈０．０５）。提示Ｐ＾５３及ｎｍ２３基因异常表达与患者预后有关，对肝细胞肝癌患者检测Ｐ＾５３和ｎｍ２３基因可预测其预后。     

P^53基因在肺癌诊治中的应用研究

Ｐ＾５３基因是一种重要的抑癌基因。在肿瘤发生时，细胞中原癌基因的激活及抑癌基因的失活导致了细胞生长的调控和分化系统的紊乱，继而发展为肿瘤细胞。Ｐ＾５３基因的点突变、缺失和来活在肺肿瘤的激发和进展过程中甚为广泛。从分子水平研究Ｐ＾５２基因的关系，对肿瘤的诊治及预后估价具有重要的临床意义，也为基因治疗提供了可靠的理论依据。     

肺癌组织中P16蛋白与血管内皮生长因子表达的临床意义

采有ＳＡＢＣ免疫组织化学方法检测了７１例肺癌患者癌组织中Ｐ＾１６蛋白和血管内皮生长因子的表达情况，并与２０例肺良性病变患者比较。结果肺癌组的Ｐ＾１６蛋白阳性北为５３．５２％，明显低于对照组，Ｐ〈０．０１；有淋巴结转移者的Ｐ＾１６蛋白阳性率显著低于无淋巴结转移者；肺癌分期为Ⅲ～Ⅳ期者Ｐ＾１６蛋白阳性率显著低于Ｉ～Ⅱ期者。肺癌组ＶＥＧＦ阳性率为７１．８３％，明显高于对照组，Ｐ〈０．０１；吸烟量少者ＶＥ     

P53抑癌基因突变产物在胆囊腺癌中的表达

Ｐ５３基因位于１７号染色体短臂上，Ｐ５３基因野生型具有肿瘤抑制作用，而具有肿瘤转化作用的是Ｐ５３基因的突变体。野生型Ｐ５３蛋白不稳定，含量低，用抗Ｐ５３蛋白单克隆抗体检测，有Ｐ５３蛋白的过量表达显示Ｐ５３基因的空变，本实验检测３４例胆囊腺癌，结果表明不同分化的腺癌都有不同程度的Ｐ５３基因突变，说明Ｐ５３基因蛋白在肿瘤早期诊断和作为预后观察是一项辅助指标。     

Aneuploidy and RAS mutations are mutually exclusive events in the development of well-differentiated thyroid follicular tumours

OBJECTIVE: Follicular thyroid tumours present several genetic alterations such as aneuploidy, RAS mutations and PAX8/PPARgammarearrangements. The molecular basis of aneuploidy remains undefined in the majority of human cancers. It has been proposed that mutations in RAS oncogenes could be related to chromosomal instability, although this issue remains controversial. The aim of our study was to investigate the correlation between aneuploidy, RAS mutations and PAX8/PPARgamma gene rearrangement in thyroid follicular tumours. DESIGN: Ploidy status was determined by flow cytometry in 111 thyroid lesions (42 follicular thyroid adenomas, 27 follicular thyroid carcinomas, 19 follicular variants of papillary thyroid carcinoma, 20 poorly differentiated thyroid carcinomas and 3 anaplastic thyroid carcinomas). RAS mutations and PAX8/PPARgamma fusion gene were investigated in 101 and 87 of these samples, respectively. RESULTS: Altogether, 12 of 50 (24%) diploid tumours presented RAS mutation which contrasts with 3 of 51 (5.9%; P = 0.0124) RAS mutations in the group of aneuploid tumours. The aneuploid tumours harbouring RAS mutations were two poorly differentiated carcinomas and one follicular variant of papillary thyroid carcinoma with poorly differentiated areas. None of the tumours with RAS mutations expressed the PAX8/PPARgamma fusion gene. Three of five (60%) follicular thyroid adenomas and 1 of 7 (14%) follicular thyroid carcinomas, with the PAX8/PPARgamma fusion gene, were aneuploid. CONCLUSIONS: Our data suggest that aneuploidy and RAS mutations are mutually exclusive events in the development of well-differentiated thyroid follicular tumours.     

Expression of P53 oncoprotein in benign and malignant lesions of large bowel

AIM: To investigate the expression of P53 oncoprotein in benign and malignant lesions of the large bowel as well as the relationship between p53 expression and clinicopathological factors. METHODS: Immunohistochemistry was used to detect P53 protein in large bowel tissues of 146 cases with benign and malignant lesions. RESULTS: All normal large bowel mucosae and non-neoplastic polyps were negative for P53 protein. However, the positive rates of P53 protein in adenomas, paracancerous mucosae and carcinomas were 18.18% (2/11), 13.21% (7/53) and 42.11% (32/76), respectively. The P53 expression in both adenomas and paracancerous mucosae presented only weak staining, whereas 75% of p53 positive cancers displayed very intense staining (++ or +++). The rates of P53 protein detection in poorly differentiated carcinoma and mucous carcinoma were 63.64% (7/11) and 62.5% (10/16), respectively, which were much higher than that of well/moderately differentiated carcinomas (30.16%, 15/40) (p < 0.05), and the carcinomas with marked positive p53 expression were more likely to penetrate the bowel wall and metastasize to lymph nodes (p < 0.05). However, no relationship between p53 expression and massive type, tumor size, location, Dukes stage or 3-year survival was found in this study. CONCLUSION: P53 gene mutation and overexpression are common in colorectal cancers, and seem to be associated with histological type, progression and lymph node metastasis of colorectal cancer.     

N-ras mutation in poorly differentiated thyroid carcinomas: correlation with bone metastases and inverse correlation to thyroglobulin expression.

Codon 61 of the N-ras oncogene was screened for mutations in 99 surgically resected thyroid carcinomas by a polymerase chain resection (PCR)-based method (PCR-primer introduced restriction with enrichment of mutant alleles [PCR-PIREMA]). A point mutation of the N-ras oncogene at the codon 61 was detected in 16 of 99 (16.2%) thyroid carcinomas examined by this method. No RAS alteration was detected in the group of 11 medullary thyroid carcinomas, while 3 of 31 (10.0%) papillary carcinomas, 2 of 5 (40%) follicular carcinomas, 8 of 44 (18.2%) poorly differentiated carcinomas, and 3 of 5 (60%) undifferentiated carcinomas showed an activation of N-RAS proto-oncogene. Interestingly, two primary follicular tumors and their corresponding bone metastases, showed N-ras mutations. In the same cases we evaluated the expression of thyroglobulin by immunohistochemical analysis. Although the majority of well-differentiated carcinomas expressed a high level of thyroglobulin, the expression of the same antigen was absent or only occasional weakly positive in 33 of 44 poorly differentiated carcinomas. Interestingly, N-ras mutation was restricted to the group of tumours with low or absent thyroglobulin expression, suggesting that this genetic change is prevalent in less differentiated tumors.     

BRAF mutations in thyroid tumors are restricted to papillary carcinomas and anaplastic or poorly differentiated carcinomas arising from papillary carcinomas

Activating point mutations of the BRAF gene have been recently reported in papillary thyroid carcinomas. In this study, we analyzed 320 thyroid tumors and six anaplastic carcinoma cell lines and detected BRAF mutations in 45 (38%) papillary carcinomas, two (13%) poorly-differentiated carcinomas, three (10%) anaplastic carcinomas, and five (83%) thyroid anaplastic carcinoma cell lines but not in follicular, Hürthle cell, and medullary carcinomas, follicular and Hürthle cell adenomas, or benign hyperplastic nodules. All mutations involved a T-->A transversion at nucleotide 1796. In papillary carcinomas, BRAF mutations were associated with older age, classic papillary carcinoma or tall cell variant histology, extrathyroidal extension, and more frequent presentation at stages III and IV. All BRAF-positive poorly differentiated and anaplastic carcinomas contained areas of preexisting papillary carcinoma, and mutation was present in both the well-differentiated and dedifferentiated components. These data indicate that BRAF mutations are restricted to papillary carcinomas and poorly differentiated and anaplastic carcinomas arising from papillary carcinomas. They are associated with distinct phenotypical and biological properties of papillary carcinomas and may participate in progression to poorly differentiated and anaplastic carcinomas.     

Do histological, immunohistochemical, and metabolic (radioiodine and fluorodeoxyglucose uptakes) patterns of metastatic thyroid cancer correlate with patient outcome?

The aim of this study is to search for relationships between histology, radioiodine ((131)I) uptake, fluorodeoxyglucose (FDG) uptake, and disease outcome in patients with metastatic thyroid cancer. Eighty patients with metastatic thyroid cancer (34 males, 46 females, mean age at the time of the diagnosis of metastases: 55 years) were retrospectively studied. All patients were treated with radioactive iodine and evaluated by FDG-positron emission tomography (PET). Primary tumor tissue sample was available in all cases. Forty-five patients (56%) had a papillary, 12 (15%) a follicular, and 23 (29%) a poorly differentiated thyroid cancer. Cellular atypias, necrosis, mitoses, thyroid capsule infiltration, and vascular invasion were frequently detected (70, 44, 52, 60, and 71% respectively). Metastases disclosed FDG uptake in 58 patients (72%) and (131)I uptake in 37 patients (45%). FDG uptake was the only significant prognostic factor for survival (P=0.02). The maximum standardized uptake value and the number of FDG avid lesions were also related to prognosis (P=0.03 and 0.009). Age at the time of the diagnosis of metastases (P=0.001) and the presence of necrosis (P=0.002) were independent predictive factors of FDG uptake. Radioiodine uptake was prognostic for stable disease (P=0.001) and necrosis for progressive disease at 1 year (P=0.001). Histological subtype was not correlated with in vivo tumor metabolism and prognosis. In conclusion, FDG uptake in metastatic thyroid cancer is highly prognostic for survival. Histological subtype alone does not correlate with (131)I/FDG uptake pattern and patient outcome. Well-differentiated thyroid cancer presenting histological features such as necrosis and FDG uptake on PET scan should be considered aggressive differentiated cancers.     

Serum vascular endothelial growth factor levels are elevated in metastatic differentiated thyroid cancer but not increased by short-term TSH stimulation

Solid tumor formation requires the development of a blood supply adequate to meet the metabolic demands of the enlarging tumor mass that cannot be sustained by simple diffusion. One principal stimulant to endothelial cell growth and migration, vascular endothelial growth factor (VEGF), is synthesized and secreted by thyroid cancer cells. Furthermore, VEGF overexpression is associated with an aggressive thyroid cancer phenotype in both animal models and clinical-pathological studies. In other malignancies, elevated serum levels of VEGF often correlate with stage of disease and other poor prognostic clinical features. Therefore, we hypothesized that serum VEGF levels would be significantly higher in patients with persistent or recurrent thyroid cancer than in those cured of the disease. Because TSH stimulates both normal and neoplastic thyroid cells, we also proposed that serum VEGF would be further increased by TSH stimulation. Sixty-nine patients with either papillary or follicular thyroid cancer, status post total thyroidectomy, and prior radioactive iodine ablation, who had undergone routine recombinant human TSH (rhTSH, Thyrogen, Genzyme Transgenics Corp., Cambridge, MA) assisted whole-body radioactive iodine scanning, were included in this study. This cohort (mean age 53 +/- 16 yr, 51% female) included 21 patients with no evidence of disease and 48 patients with local or distant metastases. Stored serum samples obtained for standard Tg determinations before and 72 h following standard rhTSH stimulation were identified and assayed for VEGF 165 (R \[amp ]\ D Systems, Minneapolis, MN). Baseline serum VEGF levels obtained at a time of TSH suppression were significantly higher in patients with known metastatic disease than in those with no evidence of disease (416 +/- 62 pg/ml vs. 185 +/- 25 pg/ml, P = 0.001). Patients with distant metastases had baseline serum VEGF levels that did not differ significantly from patients with only cervical recurrences (455 +/- 90 pg/ml in distant metastases vs. 330 +/- 44 pg/ml for local cervical recurrences). Short-term TSH stimulation, although causing a significant rise in serum Tg, resulted in no significant increase in serum VEGF measured 72 h after rhTSH injection in either the patients with known metastatic disease (416 +/- 62 pg/ml baseline vs. 419 +/- 71 pg/ml after TSH stimulation) or in cured patients (185 +/- 25 pg/ml baseline vs. 191 +/- 33 pg/ml after TSH stimulation). Subgroup analysis revealed that patients with metastatic disease arising from well differentiated primary thyroid cancers had significantly higher serum VEGF levels than patients with metastatic disease arising from poorly differentiated thyroid cancer primaries (485 +/- 74 pg/ml vs. 167 +/- 32 pg/ml, P = 0.003 by ANOVA). Poorly differentiated metastatic thyroid cancers had serum VEGF levels indistinguishable from patients cured of disease (167 +/- 32 pg/ml vs. 186 +/- 25 pg/ml). In summary, serum VEGF is significantly elevated in patients with metastatic differentiated thyroid cancer but not in those with poorly differentiated thyroid cancer metastases. No measurable increase in serum VEGF levels can be detected 72 h after short-term TSH stimulation with rhTSH. We conclude that serum VEGF may serve as a clinical useful marker of residual differentiated thyroid cancer.     

Immunohistochemical studies on the expression of P-glycoprotein and p53 in relation to histological differentiation and cell proliferation in hepatocellular carcinoma

Localization of P-glycoprotein (P-gp) and p53 was immunohistochemically examined in 41 patients with hepatocellular carcinoma (HCC) in order to determine the relationship between the expression of P-gp and p53 and the degree of histological differentiation or cell proliferation in HCC. P-gp showed different patterns of expression between cancerous and cirrhotic liver hepatocytes, and the expression in cancerous tissue also varied according to the degree of histological differentiation. In cirrhotic liver hepatocytes, expression of P-gp was found on bile canalicular membranes. In the case of cancerous tissue, P-gp was localized on the canalicular membranes in well-differentiated HCC showing a trabecular pattern, as recognized cirrhotic liver hepatocytes. In moderately differentiated HCC showing pseudo-glandular patterns, predominant expression of P-gp was found on the luminal side of cell membranes of the glandular ducts. The P-gp expression rate was 87.5% in well-differentiated HCC, 84% in moderately differentiated HCC, and 37.5% in poorly differentiated HCC, indicating a marked decrease with decreasing degree of differentiation. On the other hand, the rate of mutation of p53, a tumor suppressor gene, was 12.5% in well-differentiated HCC, 52.0% in moderately differentiated HCC, and 85.5% in poorly differentiated HCC, showing a significant increase with decreasing degree of differentiation (P<0.005). The labeling index (LI) of proliferating cell nuclear antigen (PCNA) tended to increase with the progression of chronic liver disease, with a markedly high value of 24.0±1.5% in cases of HCC. The PCNA LI was 15.6±11.9% in well-differentiated HCC, 23.1±15.1% in moderately differentiated HCC, and 50.1±13.3% in poorly differentiated HCC, which indicated a significantly increase in poorly differentiated HCC (P<0.001). Thus, it became apparent that abnormal expressions of P-gp and p53 and the cell proliferation in HCC vary according to the degree of histological differentiation of the malignancy. This suggests that more effective chemotherapy for HCC can be potentially developed by considering the pattern and level of expression of P-gp as a mechanism of drug resistance and the extent of histological differentiation.     

Lumican蛋白在胰腺导管腺癌间质中的表达及其与Ki-67、VEGF及突变型P53表达的相关性

目的:观察细胞外基质蛋白Lumican在胰腺导管腺癌(pancreatic ductal adenocarcinoma,PDA)中表达特征,分析Lumican与Ki-67、VEGF、突变型P53等肿瘤恶性表型相关分子的关联.方法:采用免疫组织化学染色(IHC)和逆转录-聚合酶链式反应(RT-PCR)检测PDA原发灶及对应癌旁胰腺组织中Lumican表达.IHC检测PDA原发灶Ki-67、VEGF及突变型P53表达.用SPSS软件行统计学分析.结果:PDA原发灶中,Lumican表达在mRNA及蛋白质水平均明显高于癌旁胰腺组织.就该蛋白在癌灶中的分布特性而言,Lumican蛋白主要定位于癌间质,阳性表达率为83.0%(83/100).低分化PDA中,癌间质过表达Lumican与TNM分期相关(x2=6.446,P<0.05),与年龄、性别、淋巴结转移、远处转移等无明显相关.高中分化PDA中,癌间质过表达Lumican与临床病理特征无关,而与Ki-67(r=-0.28,P=0.017)、VEGF(r=-0.264,P=0.025)及突变型P53(r=-0.253,P=0.032)表达呈明显负相关.结论:Lumican在PDA原发灶中表达高于癌旁胰腺组织,主要分布于癌间质.Lumican在癌间质过表达与低分化PDA的TNM分期相关,与高、中分化PDA的Ki-67、VEGF及突变型P53表达呈负相关.