结直肠肿瘤P53蛋白表达与肿瘤临床病理学特征及预后的关系

应用免疫组织化学方法观察７２例结直肠、１１例腺瘤、３０例癌旁粘主１５例正常粘膜Ｐ５３蛋白表达及其与肿瘤临床病理学特征和预后的关系。结果显示：结直肠癌Ｐ５３蛋白阳性率为５０％，腺瘤的阳性率为１８．１８％（Ｐ＜０．０５），阳性细胞主要分布在腺瘤的增生区或不典型增生区；正常粘膜、癌粘膜Ｐ５３蛋白均阴性。Ｐ５３蛋白阳性的结直肠癌多呈浸润性生长方式，且以浸润至浆膜外者居多，患者片存率较Ｐ５３蛋白阴性者（Ｐ＜     

p-MAPK、cyclinD1和p53蛋白在大肠癌中的表达及其相关性

目的：检测ｐ－ＭＡＰＫ、ｃｙｃｌｉｎＤ１和ｐ５３蛋白在大肠癌中的表达，并探讨其相关性。方法：免疫组织化学Ｓ－Ｐ法。结果：１４０例大肠癌中ｐ－ＭＡＰＫ、ｃｙｃｌｉｎＤ１和ｐ５３蛋白的阳性表达率分别为９２．９％（１３０／１４０）、８７．１％（１２２／１４０）和６６．４％（９３／１４０）；５０例相应癌旁肠粘膜中阳性表达率分别为４．０％（２／５０）、６．０％（３／５０）和２．０％（１／５０）。三者阳性表达     

大肠肿瘤中抑凋亡基因bcl－2和p53蛋白产物的表达和意义

用免疫组化方法观察４５例大肠腺瘤和６１例大肠腺癌中ｂｃ１－２和ｐ５３蛋白的表达。结果显示：正常大肠粘膜中ｂｃ１－２和ｐ５３均未见表达，而大肠腺瘤及大肠腺癌阳性率均较正常组织明显增加（Ｐ＜０．０１）．大肠腺瘤中ｂｃ１－２表达位于不典型增生较重区域。大肠腺瘤ｐ５３表达随腺瘤直径增加而增加，其中≥２０ｍｍ组阳性率显著高于＜１０ｍｍ组（ｐ＜０．０５）。ｐ５３蛋白阳性率也随不典型增生程度增加而增高。ｐ５３表达与大肠腺癌分化程度及Ｄｕｋｅｓ分期有关。大肠腺瘤中ｂｃ１－２和ｐ５３蛋白的表达呈负相关。结果表明，ｂｃ１－２蛋白表达对大肠腺瘤的增殖有一定意义，ｐ５３在大肠腺瘤癌变和大肠腺癌进展中起重要作用。     

大肠腺瘤与腺癌P21,P53,P185蛋白表达及ras,p53基因突变的检测

应用免疫组化及ＰＣＲ－ＲＦＬＰ法检测３０例大肠腺瘤、７４例大肠腺癌及癌旁粘膜Ｐ２１、Ｐ５３、Ｐ１８５蛋白表达及ｒａｓ、ｐ５３基因突变。结果表明，大肠腺瘤Ｐ２１、Ｐ５３蛋白阳性率（５３．３％，２７．６％）高于癌旁粘膜（Ｐ＜０．０１），二者过度表达与腺瘤恶性倾向有关。大肠腺癌Ｐ２１、Ｐ５３及Ｐ１８５蛋白阳性率（７２．９％，３７．８％，４７．２％）皆高于腺瘤Ｉ级不典型增生（Ｐ＜０．０１，Ｐ＜０．０５及Ｐ＜０．０５）。９例腺瘤，４０例腺癌存在两种以上蛋白表达，共同表达与腺瘤恶性倾向及腺癌预后有关。大肠腺瘤及腺癌ｒａｓ基因突变率分别为２６．７％及４１．９％，ｒａｓ基因突变与腺瘤恶性倾向有关。大肠腺瘤及腺癌ｐ５３基因２４８位点突变率分别为３．３％及１４．９％。６例腺癌存在两种基因突变，两种基因协同突变与大肠癌预后有关。提示ｒａｓ、ｐ５３及ｃ－ｅｒｂＢ－２基因改变均参与了大肠癌的发生、发展过程。     

p53基因突变与肺癌发生的早期遗传学损害

在进行肺癌与ｐ５３基因关系的系列研究过程中，发现１例支气管粘膜上皮不典型增生和１例原位癌，运用免疫组织化学方法进行ｐ５３蛋白表达检测，结果发现支气管上皮不典型增生和原位癌组织中有明显的ｐ５３蛋白阳性染色细胞，从而进一步证实了ｐ５３基因突变可能是肺癌发生的早期遗传学损害。     

c─erbB─2、ras p21及p53在大肠癌表达的免疫组化研究

对４８例大肠癌连续切片进行ｃ─ｅｒｂＢ─２、ｒａｓｐ２１及ｐ５３３种癌相关基因产物的免疫组化ＡＢＣ法及ＡＰＡＡＰ－ＩＧＳＳ双重标记研究，结果表明：（１）癌相关基因ｃ─ｅｒｂＢ─２、ｒａｓｐ２１和ｐ５３阳性表达率在癌组织分别为５０．２％（２５／４８）、４３．８％（２１／４８）和５４．２％（２６／４８）；癌旁粘膜分别为２５％（１２／４８）、１８．８（９／４８）和２０．８％（１０／４８）；而在＂正常＂结肠粘膜则分别为１２．５％（６／４８）、６．３％（３／４８）和０．（２）有２种以上癌相关基因产物同时表达者在癌组织中有２６例（５４．２％）．在癌旁粘膜有５例（１０．５％），而在正常粘膜则未见有２种癌相关基因同时表达，（３）癌组织中ｃ─ｅｒｂＢ─２＋ｐ２１、ｃ─ｅｒｂＢ─２＋ｐ５３、ｐ２１＋ｐ５３及ｃ─ｅｒｂＢ─２＋ｐ２１＋ｐ５３同时表达者分别为２．１％、１２．５％、１６．７％及２２．９％，（４）ＡＰＡＡＰ－ＩＧＳＳ双重免疫标记结果表明，单一癌细胞可同时表达２种癌基因产物。上述结果提示：ｃ─ｅｒｂＢ─２、ｒａｓｐ２１及ｐ５３可能与大肠病的发生有关，三者在大肠病的发生中可能起协同作用。     

p53基因蛋白异常表达在胃癌中的意义

目的：探讨ｐ５３基因表达异常的意义。方法：用免疫组织化学方法研究ｐ基因蛋白在胃癌中的表达。结果：７２例胃腺癌中３６例阳性，高分化腺癌和低分化腺癌Ｐ５３阳性率明显高于粘液，不同浸润深度的癌细胞与癌细胞Ｐ５３阳性程度有关，在淋巴结转移组和无转移组间Ｐ５３阳性率和阳性程度均无程度差异。病人的生存时间与Ｐ５３阳性我明显的相关性，癌旁肠上皮化生和异型增生腺体全部为阴性。结论．Ｐ５３异常 表达是细胞癌变的标志     

大肠肿瘤p53基因表达及其临床病理意义

应用免疫组织化学Ｓ－Ｐ法检测了１０６例良，恶性大肠组织中抑癌基因ｐ５３基因表达民政部结果发现；正常大肠粘膜及非肿瘤性息肉均阴性；腺癌中１８．１８％ｐ５３阳性，但均为弱阳性，局灶型，而腺癌组织中４２．１１ｐ５３阳性，其中７５％为强阳性或阳性；癌组织中ｐ５３阳性表达与大肠癌的分化、组织学类型、侵范围是淋巴结转移等因素有关，低分化腺癌及粘液癌阳性率较高，ｐ５３阳性及强阳性者更易于侵出肌层及发生淋巴结转移     

垂体腺瘤p53蛋白表达及其意义

目的：研究突变型ｐ５３癌基因在垂体腺瘤中的表达。方法：应用免疫组化ＡＢＣ法检测６４例垂体腺瘤（复发组３１例，非复发组３３例），６例正常垂体、２例垂体瘤中ｐ５３蛋白表达阳性，非复发组中有２例ｐ５３蛋白表达阳性，两组差异有显著性。２例垂体癌ｐ５３蛋白强阳性表达，６例正常垂体无阳性表达。ｐ５３蛋白在不同激素类型垂体腺癌中的阳性表达率差异无显著性。结论：ｐ５３蛋白高表达与少数垂体腺癌在潜在恶性及复发有关，     

骨肉瘤p53蛋白过度表达与预后的关系

目的；探讨ｐ５３蛋白过度表达与骨肉瘤临床病理，肿瘤细胞增殖活性及预后的关系。方法：用免疫组化方法检测５０例骨肉瘤标本中ｐ５３蛋白及增殖细胞核抗原（ＰＣＮＡ）的表达。结果５０例骨肉瘤中１６例ｐ５３蛋白阳性（３２％）；ｐ５３蛋白过度表达与骨肉瘤组织学类型，组织学分级及ＰＣＮＡ分级无关，１６例ｐ５３蛋白阳性病你中，１０例在２年内死亡（６２．５％）３４例阴性病例中，１１例在２年内死亡（３２．４％），结论；     

Concurrent p53 expression in bronchial dysplasias and squamous cell lung carcinomas.

We analyzed the p53 protein immunohistochemically in bronchial dysplasias or squamous cell carcinomas in situ and in squamous cell lung carcinomas occurring in the same patients. The polyclonal antibody used (CM-1) is directed against the wild-type p53 protein, but also recognizes the mutated p53 in formalin-fixed and paraffin-embedded sections. To study the integrity of basement membranes (BMs) and the possible invasion of the dysplastic epithelium, immunostainings for the BM proteins laminin and type IV collagen were used. Nine of the 17 dysplasias showed p53 protein expression (53%); it was significantly more often seen in severe dysplasias and carcinomas in situ than in mild or moderate dysplasias (P = 0.04). The p53 antigenicity was generally located in the basal part of the epithelium. The BMs beneath mildly dysplastic epithelia were continuous. In contrast, those under moderately or severely dysplastic epithelia showed occasional disruptions. p53 protein expression was also found in dysplastic epithelium above a continuous BM suggesting an ominous process before signs of invasion. Twelve of the 17 squamous cell carcinomas showed p53 protein expression (71%). There was a significant concurrent p53 expression in bronchial dysplasias and their related squamous cell carcinomas (P = 0.009), so that all nine cases of p53 positive bronchial dysplasia also showed p53 positivity in the associated squamous cell carcinomas. These findings indicate that p53 protein expression is possible in premalignant bronchial lesions, and suggests that the p53 expression could, at least in some cases, be an early event in the development of a squamous cell carcinoma of the lung.     

Angiogenesis in preinvasive, early invasive bronchial lesions and micropapillomatosis and correlation with EGFR expression

AIMS: To study the association between morphological changes of the bronchial epithelium and its angiogenic status evaluated by microvessel count (MVC), in order to gain a better understanding of bronchial carcinogenesis. Also, to correlate MVC with epidermal growth factor receptor (EGFR) expression. METHODS AND RESULTS: Eighty-three biopsy specimens were assessed for MVC: four normal bronchial epithelia, 23 hyperplasias, 26 metaplasias, two mild dysplasias, five moderate dysplasias, nine severe dysplasias, three carcinomas in situ, six early invasive squamous cell carcinomas (EIC) and five cases of micropapillomatosis. We observed a statistically significant difference in terms of MVC between EIC and all other subgroups and between micropapillomatosis and all other subgroups. There was also a statistically significant difference between micropapillomatosis and EIC. We did not observe any difference in MVC between normal mucosa, metaplasias, hyperplasias, dysplasias or carcinoma in situ. EGFR expression was higher in severe dysplasia, carcinoma in situ and EIC, whereas it was very low in micropapillomatosis. A statistically significant difference was observed in the expression profile of EGFR vs. MVC. EGFR expression was increased in severe dysplasia, whereas an increase in MVC occurred only in EIC. CONCLUSION: During bronchial carcinogenesis, except for micropapillomatosis, EGFR expression appears to be a prerequisite for neoangiogenesis in bronchial carcinogenesis.     

p53 expression in normal,dysplastic, and neoplastic laryngeal epithelium. Absence of a correlation with prognostic factors

p53 expression has been examined in 89 squamous cell carcinomas of the larynx (34 glottic, 28 supraglottic, 18 transglottic, 8 pyriform sinus, and 1 subglottic) obtained from 88 patients surgically treated in our centre. In addition, 59 laryngeal samples including normal respiratory epithelium and non-invasive squamous cell lesions were also tested. Frozen sections were immunostained with PAb 1801 and the results were correlated with pathological features, DNA ploidy and S-phase of the tumours, disease-free interval, and survival of the patients. p53 immunoreactivity was observed in 57 (64 per cent) carcinomas. None of the eight samples of normal respiratory epithelium was positive. p53-positive cells were seen in 8 of 23 (35 per cent) squamous cell metaplasias, 6 of 19 (32 per cent) low-grade dysplasias and 5 of 10 (50 per cent) high-grade dysplasias. No correlation was found between p53 expression in carcinomas and their clinical and pathological characteristics, DNA ploidy, or proliferative activity. Neither disease-free nor overall survival showed differences between p53-positive and p53-negative cases. These findings indicate that p53 may play a role in an early stage of malignant transformation of a subset of squamous cell carcinomas of the larynx, but seems not to be associated with further progression of the tumours.     

Expression of minichromosome maintenance 2 (MCM2), Ki-67, and cell-cycle-related molecules, and apoptosis in the normal-dysplasia-carcinoma sequence of the oral mucosa.

OBJECTIVE: We examined cell cycle and cell death biomarker trends with the normal-dysplasia-carcinoma sequence of the oral epithelia analyzing the pathological significance of a new biomarker, minichromosome maintenance 2 (MCM2). METHODS: This study analyzed 12 patients with normal oral epithelia, 69 with dysplasia, and 35 with squamous cell carcinoma (SCC); in 13 patients, SCCs were preceded by dysplasia. The sections were immunostained for MCM2, Ki-67, P53, P27(Kip1) and P21(CIP1/WAF1), and conducted by TUNEL methods. Western blot analysis of MCM2 was performed in the 4 human cultured oral SCCs, all of which showed the expression. RESULTS: Significantly higher labeling indices (LI; %) of MCM2, Ki-67, and P53, as well as lower LI of TUNEL indices (TI; %), P27, and P21 were noted in the SCCs than in the dysplasias. The 13 dysplasias developed SCC with significantly higher LI of MCM2 and P53, and lower LI of P21 than the other dysplasias (each p < 0.05). The LI of MCM2, P21 and the TI were not correlated with P53 expression. CONCLUSIONS: Oral dysplasia was characterized by lower cell proliferation and a higher frequency of cell death compared to SCCs. The higher LI of MCM2 and P53 and the lower LI of P21 might predict malignant transformation of oral dysplasia. MCM2 is regulated via a P53-independent pathway, and a useful biomarker of proliferating cells.     

p53 GENE ALTERATIONS AND p53 PROTEIN IN ORAL EPITHELIAL DYSPLASIA AND SQUAMOUS CELL CARCINOMA

To examine the expression of p53 protein and gene alterations in oral epithelial lesions including epithelial dysplasias and primary squamous cell carcinomas, immunohistochemical and temperature gradient gel electrophoresis (TGGE) methods were applied to formalin-fixed and paraffin-embedded tissues. Morphologically normal mucosal epithelium stained negatively for p53 protein. Three out of 11 (27·3 per cent) epithelial dysplasias and 19 out of 57 (33·3 per cent) primary squamous cell carcinomas stained positively for p53 protein. Although more than half of the cases were positive for p53 protein in stage I, the positive cancer cases were found at other stages with variable frequency. Immunoreactive products were localized in the nucleus, especially in the basal and suprabasal layers. The analysis by TGGE revealed gene alterations in exons 5–8 in 3 out of 3 epithelial dysplasias and 17 out of 19 (89·5 per cent) primary squamous cell carcinomas which were immunohistochemically positive for p53 protein. These results suggest that p53 gene mutation may be involved in carcinogenesis in the oral squamous epithelium even in the early stage of the dysplasia–carcinoma sequence.     

Immunohistochemical expression of metallothionein in benign premalignant and malignant epithelium of the larynx: correlation with p53 and proliferative cell nuclear antigen

In this study we evaluated the immunohistochemical expression of metallothionein (MT) in 44 squamous cell carcinomas, 14 cases of in situ carcinoma, 47 with epithelial dysplasia, 11 papillomas and 21 cases of keratosis. The MT expression was studied in correlation with p53 protein expression and the proliferative cell nuclear antigen (PCNA). The monoclonal antibodies E9 (anti-MT), DO-7 (which reacts with a denaturation-resistant epitope in wild-type and mutant p53) and PC10 (anti-PCNA) on formalin-fixed, paraffin-embedded tissue were used employing the immunoperoxidase (ABC) method. The immunohistochemical localization of MT has shown its rather ubiquitous presence in the cytoplasm and nucleus of both benign and malignant epithelial cells. In most cases the adjacent "normal" epithelium showed low positivity in the basal portion. The mean value of metallothionein expression was 35.73 in squamous cell carcinomas, 32.21 in in situ carcinomas, 11.86 in dysplastic epithelium, 5.10 in papillomas and 3.5 in keratosis. In carcinomas, low MT expression (< 10% of neoplastic cells) was observed in 20.5% of the cases, moderate (10%-50% of neoplastic cells) in 54.5% and extensive expression (> 50% of neoplastic cells) in 25% of the cases. We did not find any statistically significant difference of MT expression between in situ and infiltrating carcinomas, while we did observe a significant difference between carcinomas and the other groups. There was a statistically significant difference in the PCNA values in both benign and malignant lesions, while no statistically significant difference was observed in p53 protein expression in the above groups. A positive correlation between MT expression and the PCNA value (p < 0.0001) in the benign and malignant groups was detected. The PCNA value was also correlated with the p53 protein expression (p = 0.001). No correlation was found between MT and p53 protein expression. In conclusion, these results suggest that the MT expression may play a role in the development of malignant disease of the larynx, from the early phase of laryngeal carcinogenesis, independently from the p53 expression. It is also possible to contribute to tumour cell growth, as determined by the PCNA score.     

p16 Alterations and retinoblastoma protein expression in squamous cell carcinoma and neighboring dysplasia from the upper aerodigestive tract

The progression potential of preinvasive epithelial lesions is usually evaluated by assessing the degree of histologic dysplasia. We examined p16, retinoblastoma protein (pRb), and proliferating cell nuclear antigen (PCNA) immunophenotypes in 57 cases of previously untreated squamous cell carcinoma (SCC) of the upper digestive tract and in the neighboring normal and dysplastic epithelia. Tissue samples were examined for homozygous deletion of exon 2 of the p16 gene using polymerase chain reaction (PCR) analysis. The PCNA index increased with increasing grade of dysplasia. The pRb protein was expressed in 89% of the samples of SCCs and in the neighboring dysplasias and carcinoma in situ (CIS). In cases with a lack of pRb expression, corresponding preinvasive lesions were also negative. Lack of p16 expression was found in 82% of SCCs. The prevalence of p16 expression decreased with increasing grade of dysplasia. Molecular analysis of the p16 gene showed homozygous deletion in 37% of SCCs, 33% of CIS, and 15% of the samples of normal epithelia. Our data indicate that inactivation of p16 may play an important role in early head and neck carcinogenesis, whereas the mutation of Rb may be an infrequent event. The p16 immunophenotype might be a biomarker for an increased risk of progression in squamous dysplasia.     

HPV type 16 in conjunctival and junctional papilloma, dysplasia, and squamous cell carcinoma.

AIMS--To clarify the role of human papillomavirus (HPV) infection in the development of papilloma, dysplasia, squamous cell carcinoma, and basal cell epithelioma arising from the eyelids, including the tunica conjunctiva palpebrum (conjunctiva), its junction to epidemis of eyelid skin (junction), and eyelid skin. METHODS--Sixteen cases of papilloma, four of dysplasia, four of squamous cell carcinoma, and 12 of basal cell epithelioma were examined using formalin fixed and paraffin embedded samples. Detection of HPV-DNA was performed by PCR-RFLP and in situ hybridisation (ISH) methods. RESULTS--HPV-16 was detected in 12/16 papillomas (75%), 2/4 dysplasias (50%), and 1/4 squamous cell carcinomas (25%) but in none of the basal cell epitheliomas. No other HPV subtypes were found. ISH assay showed positive signals in only two cases of dysplasia and squamous cell carcinoma. The mean age of HPV-16 positive dysplasia and squamous cell carcinoma cases (81.7 years) was significantly higher than that of HPV-16 positive papilloma cases (p < 0.01). CONCLUSIONS--Based on the presence of HPV-16 in both benign and malignant lesions and the age distribution, it seems likely that HPV-16 alone may be incapable of causing development of conjunctival and junctional dysplasia and squamous cell carcinoma, and that any correlation between the papilloma-squamous cell carcinoma sequence and HPV infection may be due to rare events.     

Divergent expression of L-type amino acid transporter 1 during uterine cervical carcinogenesis

L-type amino acid transporter 1 (LAT1) is a Na?-independent neutral amino acid transporter that has an essential role in cell proliferation. Although LAT1 expression is observed in various tumor cell lines and immunohistochemical expression of LAT1 has been investigated in carcinomas of various organs, LAT1 expression in uterine cervical neoplasm has not been reported. Therefore, in the present study, we immunohistochemically analyzed LAT1 expression along with the well-known markers of cervical carcinogenesis Ki-67 and p16 in normal uterine cervical mucosa (49 specimens) as well as cervical intraepithelial neoplasia (17 mild or moderate dysplasias and 19 severe dysplasias or carcinomas in situ) and invasive carcinomas (17 squamous cell carcinomas and 9 adenocarcinomas). LAT1 expression was limited to the basal layer of normal squamous epithelium, and it was significantly decreased in cervical intraepithelial neoplasia (P < .001), generally paralleled by increased expression of Ki-67 and p16. Interestingly, in invasive squamous cell carcinoma, LAT1 expression again increased especially at the invasive fronts (P < .001), whereas Ki-67 and p16 expressions were almost unchanged relative to noninvasive neoplasia. Although virtually no LAT1 expression was demonstrated in normal uterine cervical glands, LAT1 expression was observed in some adenocarcinomas (P < .001). The present study suggests that LAT1 expression decreases because of human papillomavirus infection as reflected by p16 overexpression in cervical intraepithelial neoplasia, whereas LAT1 expression in invasive carcinoma is associated with acquired malignant potential.     

Reduced p21(WAF1/CIP1) expression is an early event in gallbladder carcinogenesis and is of prognostic significance for patients with carcinomas of the gallbladder

The molecular basis underlying the development and progression of gallbladder carcinoma (GBC) remains poorly understood. To evaluate the roles of p21(WAF1/CIP1) and p53 in gallbladder carcinogenesis and to assess their prognostic significance for patients with GBC, we used immunohistochemistry to examine the expression of p21(WAF1/CIP1) and p53 protein in a series of surgically resected specimens, including normal epithelia, precancerous lesions adenoma, and dysplasia, and carcinomas of the gallbladder. Reduced p21(WAF1/CIP1) expression was frequently observed in carcinomas (18 of 37 lesions; 49%), and even in precancerous lesions adenomas (3 of 7; 43%) and dysplasias (5 of 5; 100%). p53 overexpression was detected in 43% of the adenomas, 60% of the dysplasias and 57% of the carcinomas. There was an inverse relationship between p21(WAF1/CIP1) and p53 expression in GBCs (P =.01). Survival analysis indicated that reduced p21(WAF1/CIP1) expression was significantly associated with shortened disease-free and overall survival (P =.04 and.03, respectively) for patients with stages II to IV GBCs. These observations suggest that reduced p21(WAF1/CIP1) expression and p53 overexpression contribute to GBC from an early stage and that determination of p21(WAF1/CIP1) expression in surgically resected specimens would add prognostic information to conventional pathologic examinations for patients with advanced-stage GBC.