Expression of p53 protein related to human papillomavirus and DNA ploidy in superficial esophageal carcinoma

We examined the p53 protein and human papilloma virus (HPV) by immunohistochemistry and DNA ploidy by cytofluorometry in paraffin-embedded esophageal carcinoma tissue specimens. Sixty-one patients with superficial esophageal carcinoma were operated on between 1983 and 1991 without any prior treatment. Immunostaining of the anti-p53 protein antibody (CM1) was positive in 32 carcinomas (52%). Patients with p53-positive tumors had a poorer outcome than those with p53-negative tumors (P<0.05). In addition, patients with p53-positive tumors did not have any characteristic site of relapse. Only 5 of the 61 patients (8.2%) had HPV-positive tumors. One of these 5 carcinomas expressed both p53 protein and HPV. Three patients with HPV-positive tumors which had invaded the submucosal layer died of relapse. A determination of DNA ploidy revealed 30 patients with aneuploid tumors, 13 with polyploid tumors and 18 with diploid tumors. The outcome of the patients with aneuploid tumors was worse than that of the patients with diploid tumor (P<0.05). p53 protein expression was not associated with DNA ploidy; however, the 16 patients who had both p53-positive and aneuploid tumors had a worse prognosis than patients with p53-negative and aneuploid tumors (P<0.01). These findings suggest that p53 protein expression in conjunction with DNA ploidy may be a useful indicator in evaluating the prognosis of patients with superficial esophageal carcinoma.     

The relation of p53 protein nuclear accumulation and angiogenesis in human prostatic carcinoma

All neoplasms require angiogenesis and resulting neovascularity for growth beyond 1 mm(2). Quantitative microvessel density (MVD) has been shown to provide staging and prognostic significance in human prostate cancer (CaP). recently, it has been demonstrated that loss of the wild-type allele of the p53 tumour suppressor gene results in reduced expression of thrombospondin-1 (TSP-1), a potent inhibitor of angiogenesis. There is also an increased expression of vascular endothelial growth factor which promotes neovascularization. p53 gene mutation and MVD were investigated in men with prostate cancer. Sections from 103 radical prostatectomy cases were evaluated with immunohistochemistry to detect mutant p53 proteins. Quantitative MVD was performed on the cases exhibiting p53 positive staining and compared with negative fields of similar Gleason grade on the same histologic sections. Twenty of the 103 cases (19.4%) revealed positive p53 staining nuclei. In 19 of these 20 cases, the MVD in p53 positive areas was greater than corresponding control regions (overall P<0.0001). Extent of p53 abnormality, as well as MVD, correlated with pathologic stage. These data suggest that mutations of the p53 tumour suppressor gene may be associated with increased angiogenesis in CaP. In addition to providing staging and prognostic information, this relationship potentially has therapeutic implications.     

Detection of human papillomavirus DNA and p53 gene mutations in human prostate cancer

The relationship between integration with human papillomavirus (HPV) and p53 gene mutations in tissues of prostate cancer was examined. Tissue samples analyzed were obtained by total prostatectomy (29 stage B cancer cases) and from autopsy (22 endocrine therapy-resistant metastatic disease cases). HPV DNA was detected in 8 of 51 (16%, 5 in stage B and 3 in autopsy cases) by polymerase chain reaction (PCR) using consensus primers on L1 region. Genotypes of HPV were entirely type 16. Structural abnormalities of p53 gene were detected in 7 of the 22 autopsy cases (32%) by PCR-single-strand conformation polymorphism analysis and direct sequencing. No p53 gene mutation was found in stage B cancer cases. Analysis of mutation spectra revealed clear differences between Japanese and Westerners. There was a significant difference in the mutation frequency between stage B and autopsy cases (P < 0.01, Fisher's exact test). One case showed both integration of HPV and p53 gene mutation in different cancer foci. However, the other cases revealed an inverse correlation between the presence of HPV DNA and p53 gene mutations. These data show that p53 genetic alteration is correlated with the progression of prostate cancer, in contrast to the integration of HPV that may occur in a relatively early stage. In conclusion, this study may indicate that either p53 gene mutation or the presence of HPV's oncogenic protein E6 is involved in the development of prostate cancer. © 1996 Wiley-Liss, Inc.     

Human papillomavirus and p53 protein immunoreactivity in condylomata acuminatum and squamous cell carcinoma of penis

Aim: To determine the immunoreactive pattern of human papillomavirus (HPV) antigen and p53 protein in condylomata acuminatum (CA) and squamous cell carcinoma (SCC) of penis. Methods: Immunohistochemistry for HPV and p53 were performed in 40 specimens of formalin fixed, paraffin embedded tissues using a polyclonal (rabbit) antibody against HPV and a monoclonal (mouse) antibody against human p53 protein. Twenty one cases of CA and nineteen cases of SCC were examined. Results: HPV antigen was detected in all 21 CA and 2 penile SCC. p53 protein overexpression was observed in 12 of 19 (63%) SCC in which 6 cases were strong positive. Five of 21 CA (24%) showed low-grade p53 protein overexpression. Conclusion: CA is related to HPV infection and some cases show p53 protein low-grade overexpression. In contrast, p53 protein overexpression is common in penile SCC, which is seldom related to HPV infection.     

乳腺癌中p53基因DNA水平突变与核蛋白水平突变的比较

目的研究ｐ５３基因ＤＮＡ水平和蛋白水平的突变。方法分别采用多聚酶链反应单链构象多态分析银染方法和链酶亲和素生物素过氧化物酶复合物（ｓｔｒｅｐａｖｉｄｉｎｂｉｏｔｉｎｐｅｒｏｘｉｄａｓｅｃｏｍｐｌｅｘ,ＳＡＢＣ）免疫组化方法,检测了５４例乳腺浸润性导管癌标本。结果２２例在ＤＮＡ水平突变阳性,突变率为４１％;３２例在核蛋白水平突变,突变率５７％。其中２２例ＤＮＡ突变阳性者,核蛋白突变均阳性;而３２例ＤＮＡ水平突变阴性者,核蛋白突变阳性者９例,占２８％。ｐ５３ＤＮＡ突变与核蛋白突变之间有显著相关性,并且两者都与患者的淋巴结转移以及雌、孕激素受体表达有关（Ｐ＜００１）。结论免疫组化是一种快速检测ｐ５３是否存在异常的有效方法。但是在蛋白水平突变阳性者中存在一定的假阳性。ｐ５３突变阳性患者预后较差     

Increased p53 protein expression in human failing myocardium.

BACKGROUND: The p53 gene is a tumor-suppressor gene which involves apoptosis and cell-cycle arrest under certain stress stimulate. However, the status of the p53 gene expression in human myocardium in congestive heart failure (CHF) remains unclear. Therefore, the current study was designed to investigate the expression of the p53 protein in human myocardium in normal subjects and in patients with severe CHF. METHODS: Human ventricular cardiac tissue was obtained from 7 normal subjects and 7 end-stage CHF patients during cardiac transplantation. The expression of p53 protein was determined by immunohistochemical staining. The cardiac apoptosis was determined by TUNEL staining. RESULTS: The p53 protein was minimally stained in normal human ventricular cardiomyocytes. In contrast, the staining density and positive stained nuclear (%) of p53 was significantly increased in ventricular cardiomyocytes of patients with severe CHF. Apoptosis in CHF human myocardium also markedly increased. CONCLUSIONS: The significantly increased expression of p53 in CHF human cardiomyocytes suggests that p53 may play an important pathophysiological role in the process of CHF through mechanisms involving myocardial apoptosis.     

Presence of human papillomavirus 6/11 DNA in condyloma acuminatum of the urinary bladder.

A Japanese woman with condyloma acuminatum of the urinary bladder is presented. The condyloma acuminatum lesion was resected endoscopically and human papillomavirus 6/11 DNA was detected. After treatment, there has been no recurrence of the disease.     

高危性人乳头瘤病毒与抑癌基因P53在膀胱癌中的表达

用ＰＣＲ法检测５２例膀胱癌组织中人高危性乳头瘤病毒（ＨＰＶ）ＤＮＡ，同时用免疫组化方法检测其Ｐ５３蛋白的过度表达，发现２８例（５３．８４％）ＨＰＶＤＮＡ阳性，１９例（３６．５４％）肿瘤细胞核Ｐ５３染色阳性，二者同时阳性者３例（５．７７％）。ＨＰＶＤＮＡ阳性者多见于高分化、非侵袭性肿瘤中，相反Ｐ５３蛋白阳性则主要表现在低分化、侵袭性肿瘤中。ＨＰＶＤＮＡ和Ｐ５３的表达呈显著性负相关（ｒ＝－０．５７６９）。结果提示：ＨＰＶ感染或Ｐ５３的过度表达与膀胱肿瘤的生物学行为有关，并可将其作为膀胱肿瘤的预后评价指标。     

Immunoexpression of p53 protein and proliferating cell nuclear antigen in penile carcinoma.

PURPOSE: We examined p53 protein and proliferating cell nuclear antigen immunoexpression as prognostic factors to the outcome of squamous cell carcinoma of the penis in 50 patients. MATERIALS AND METHODS: Penectomy and lymphadenectomy were performed in 14 patients with clinically positive nodes while 36 with cN0 disease were treated with penectomy and kept under surveillance that resulted in subsequent lymphadenectomy due to nodal relapse in 8. Of 21 patients with confirmed nodal metastases 18 died of disease. Immunohistochemical reactions were performed via the avidin-biotin-immunoperoxidase method and the results were compared with tumor pT stage, grade, nodal status and cause specific death. RESULTS: In univariate analysis proliferating cell nuclear antigen staining showed association only with nodal metastasis (p = 0.04) while p53 staining exhibited correlation with tumor pT stage (p = 0.0005), grade (p = 0.02), lymphatic spread (p = 0.02) and cause specific survival (p = 0.003). Multivariate analysis showed that p53 immunoreactivity was the only factor with prognostic significance for disease progression and cause specific survival. Tumor pT stage, grade and proliferating cell nuclear antigen staining had no significance for nodal metastases and cause specific death. CONCLUSIONS: Proliferating cell nuclear antigen staining had no prognostic value for disease progression. Since p53 over expression was associated with tumor progression and cause specific death, perhaps it should be evaluated in staging and therapeutic planning for patients with squamous cell carcinoma of the penis.     

FHIT和p53在结肠癌中的表达及其意义

目的探讨脆性组氨酸三联体（FHIT）基因和p53基因在结肠癌中的表达及其与临床病理因素的关系。方法采用免疫组织化学链菌素亲生物素一过氧化酶法（SP法）法检测74例结肠癌组织中FHIT和p53的表达。结果FHIT和p53在结肠癌中表达率分别为52．7％和56．8％。FHIT蛋白的丢失在结肠癌中占47．3％,FHIT蛋白低表达癌在癌组织学分级中的分布为低分化癌10／12,高中分化癌25／62,;在Dukes分期中的分布为C、D期癌23／37,A、B期癌12／37;在有淋巴转移组中为23／37,无淋巴转移组中为12／37。30例获访病例中,FHIT蛋白低表达癌在5年随访病例组中的分布为5年生存组为6／17,5年死亡组为10／13。两者与结肠癌的分化程度、病理分期（Dukes分期）、淋巴转移情况和五年生存率均有明显相关（P〈0．05）。结论结肠癌组织中FHIT蛋白的丢失是频发事件,FHIT可能是结肠癌发生中重要的抑癌基因;FHIT和p53的表达。可作为评估结肠癌高危因素和恶性生物学行为的参考指标。     

Human papillomavirus and p53 mutational status as prognostic factors in head and neck carcinoma

BACKGROUND: Mutations of the p53 tumor-suppressor gene are common in squamous cell carcinoma of the head and neck (SCCHN) and may portend a worse prognosis. Human papillomavirus (HPV) represents another potential prognostic factor for SCCHN. The oncogenic potential of HPV may be due to the ability of its E6 oncoprotein to promote degradation of wild-type p53 protein. We wish to determine whether there is a lower incidence of p53 mutations in HPV-positive versus HPV-negative tumors, and if HPV and/or p53 status has an impact on survival. METHODS: Thirty-two SCCHN specimens were analyzed for mutations of the p53 gene using single-strand conformational polymorphism (SSCP) analysis followed by DNA sequencing. The HPV status of all specimens was evaluated by use of polymerase chain reaction with HPV consensus primers and Southern blot hybridization. Pertinent clinical information was obtained from chart review. RESULTS: Nonsilent p53 mutations were present in 2 of 15 (13%) of HPV-positive tumors compared with 6 of 17 (35%) of HPV-negative tumors (p =.229; Fisher's exact test, odds ratio.28). A survival advantage was found between HPV-positive compared with HPV-negative specimens (p =.0264) and between p53 wild type compared with p53 mutant specimens (p =.01) by univariate log rank analysis. When stratified according to both HPV and p53 status, a statistically significant survival difference was observed largely because of a 100% survival for the HPV-positive/p53 wild-type group (p =.003). CONCLUSIONS: This preliminary study supports the notion that the presence of HPV confers a survival advantage among HNSCC patients, particularly when p53 is wild type.     

Detection of human papillomavirus DNA and expression of p16, Rb, and p53 proteins in small cell carcinomas of the uterine cervix

Human papillomavirus (HPV) has been implicated as an etiologic agent for the development of primary small cell carcinoma of the uterine cervix, a rare but highly aggressive malignancy. It has been shown that the HPV E6 and E7 oncoproteins are able to inactivate the tumor suppressor functions of p53 and Rb. In squamous cell carcinoma and adenocarcinoma of the cervix, HPV infection is also associated with overexpression of p16, a cyclin-dependent kinase inhibitor. In this study, 22 cases of primary small cell carcinoma of the uterine cervix were subjected to broad-spectrum HPV DNA amplification and typing, and immunohistochemically examined for the expression of p16, Rb, and p53 proteins. The results show that HPV DNA was detected in every case (100%), with 18 cases (82%) harboring type 18. The tumor cells exhibited strong nuclear staining for p16 in 20 cases (91%). This was associated with a complete loss of Rb nuclear staining in tumor cells in 16 cases (73%). The p53 protein was essentially undetectable in all cases. In contrast, HPV DNA was not detected in 9 colorectal and 8 urinary bladder small cell carcinomas included in this study for comparison. While similar p16 and Rb expression patterns were observed in these HPV-negative tumors, a different expression pattern for p53 was noted where strong nuclear staining was seen in 8 cases (47%; P = 0.0004 compared with cervical tumors). These observations indicate that different mechanisms are involved in the pathogenesis of small cell carcinomas of the uterine cervix and support the notion that nuclear p16 overexpression serves as an indication of Rb defunctioning in tumor cells, which may or may not result from high-risk HPV infection.     

Expression of p53 product in Chinese human bladder carcinoma

Summary Expression of p53 protein was examined in 67 cases of primary transitional cell carcinoma of the bladder and 6 normal controls using an immunohistochemical method on paraffin sections. Positive nuclear staining for p53 in malignant cells was found in 34 (51%) of the 67 cancer patients; no positive staining for p53 was detected in any of the normal controls or in the benign cells, including stromal and inflammatory cells, within the tumor tissue. There were 8 positive cases (33%) in 24 grade G1 tumors, 12 (48%) in 25 G2 tumors and 14 (78%) in 18 G3 tumors. p53 protein was detected positively in 14 (36%) of 39 superficial tumors (Tis-T1) and in 20 (71%) of 28 invasive tumors (T2–T4). Thus, positive staining for p53 was found more frequently in poorly differentiated tumors (chi-squared test: G3/G1+G2P<0.01) and in invasive tumors (chi-squared test: T2–T4/Tis-T1P<0.01). Expression of p53 was also closely associated with recurrence of tumors. Alterations in p53 expression may be of prognostic value in cases of bladder transitional cell carcinoma.     

Expression of mdm-2 and p53 protein in transitional cell carcinoma

Amplification of the mdm-2 gene and overexpression of the mdm-2 protein might inactivate p53 function, and may have prognostic relevance. The present paper investigated the immunohistochemical overexpression of the mdm-2 and p53 proteins in 25 biopsy specimens of transitional cell bladder carcinomas (10 pT1 and 15 pT2 or higher stages). Five cases (20%) showed strong mdm-2 protein immunoreactivity in more than 5% of the tumor cells; 14 cases (56%) showed p53 immunoreactivity in more than 20% of the cells, and were considered as overexpressing p53 protein. Four of the five cases with strong mdm-2 immunoreactivity did not show p53 overexpression, and 13 of the 14 cases with p53 overexpression did not show mdm-2 immunoreactivity. Our data are consistent with the hypothesis that p53 overaccumulation (and hence possible p53 gene mutation) or mdm-2 overexpression (and hence possible mdm-2 gene amplification) may mirror two different and possibly complementary gene alterations, which might finally interfere with the control of cell proliferation and apoptosis. In this perspective, evaluation of the combined mdm-2/p53 protein phenotype in human bladder carcinomas could have prognostic relevance and give us better prognostic information than evaluation of the p53 protein alone.     

阴茎癌组织中人乳头瘤病毒DNA的原位杂交研究

为了研究人乳头瘤病毒与阴茎癌的关系，应用地高辛标记的ＨＰＶ６、ＨＰＶ１１、ＨＰＶ１６和ＨＰＶ１８ＤＮＡ探针分别对４６例阴茎癌组织进行原位杂交检测ＨＰＶＤＮＡ。结果显示：ＨＰＶＤＮＡ阳性２２例，其中ＨＰＶ１６ＤＮＡ阳性２０例，ＨＰＶ１８ＤＮＡ阳性４例，在２例转移癌和癌旁不典型增生组织中检测到了ＨＰＶ１６ＮＤＡ，未见ＨＰＶ６ＤＮＡ和ＨＰＶ１１ＤＮＡ阳性，ＨＰＶＤＮＡ位于癌细胞核中。结果表明原位杂交可以用来研究阴茎癌组织中ＨＰＶ的存在及分型，同时也证实了阴茎癌的发生和ＨＰＶ１６及ＨＰＶ１８感染有密切关系     

乳腺癌患者高危型人乳头状瘤病毒感染、p53蛋白表达及淋巴结转移分析

目的分析乳腺癌患者高危型人乳头状瘤病毒（HPV）感染、p53蛋白表达和淋巴结转移率。 方法选取2016年5月至2018年5月于西北妇女儿童医院治疗的乳腺癌患者共90例作为观察组,选取同期于本院治疗乳腺增生患者90例作为对照组。对癌组织行HPV基因分型和p53蛋白检测,增生组织行HPV基因分型。观察入组患者高危型HPV感染率,分析乳腺癌患者肿瘤大小、TNM分期、淋巴结转移率与p53蛋白表达等。 结果观察组患者中共60例发生高危型HPV感染,对照组中共36例发生高危型HPV感染,观察组患者中HPV16、18基因型感染率分别为21.11%（19/90）和22.22%（20/90）,均高于对照组[2.22%（2/90）和2.22%（2/90）],差异均有统计学意义（χ² = 7.108、P = 0.001,χ² = 8.063、P = 0.001）。观察组HPV阳性患者淋巴结转移率为93.33%（56/60）,显著高于HPV阴性患者（21/30、70.00%）,差异有统计学意义（χ² = 4.072、P = 0.002）。观察组患者中p53蛋白阳性者39例（39/90、43.33%）,其中肿瘤大小≤ 2 cm者27例（27/39、69.23%）、TNM分期Ⅰ期、Ⅱ期、Ⅲ期和Ⅳ期分别占15.38%（6/39）、30.77%（12/39）、35.90%（14/39）和17.95%（7/39）,淋巴结转移率为82.05%（32/39）;p53蛋白阴性患者51例（51/90,56.67%）,肿瘤大小≤ 2 cm者35例（35/51、68.63%）、TNM分期Ⅰ期、Ⅱ期、Ⅲ期和Ⅳ期分别占7.84%（4/51）、19.61%（10/51）、49.02%（25/51）和23.53%（12/51）,淋巴结转移率为86.27%（44/51）,p53蛋白阴性组与p53蛋白阳性组患者肿瘤大小、TNM分期及淋巴结转移率差异均无统计学意义（χ² = 0.682、P = 0.462,χ² = 0.491、P = 0.507,χ² = 0.572、P = 0.461）。 结论检测乳腺癌患者高危型人乳头状瘤病毒感染、p53蛋白表达及淋巴结转移率有助于对乳腺癌的诊疗;p53蛋白阳性率下降可能促进HPV感染相关肿瘤的发生和发展。     

No evidence of human papillomavirus DNA in breast carcinoma in Tunisian patients

The aim of this study was to evaluate the prevalence of broad range of anogenital HPVs in a series of 123 Tunisian breast carcinoma cases. PCR assays were performed to amplify regions within the L1, E1, E6 and E7 open reading frames of a broad range of anogenital HPVs and specific types HPV16, 18, 31 and 33. In addition, we performed an in situ hybridization analysis using HPV biotinylated DNA probes for the detection of broad spectrum of anogenital HPV types, high-risk HPV types (16 and 18), intermediate-risk HPV types (31 and 33) and low-risk HPV types (6 and 11). None of the 123 breast carcinoma samples showed PCR amplification of HPV DNA using the broad spectrum consensus primer-pairs E1-350L/E1-547R and GP5+/GP6+ primers. Furthermore, neither high risk nor low-risk HPV types were detected in any of these cases. Moreover, using in situ hybridization for the detection of HPVs, we failed to detect a positive signal in neoplastic cells in any case. Our results suggest that anogenital papillomaviruses are unlikely to play a role in the development of breast carcinomas in Tunisian patients.     

Expression of p16(INK4A), p53, and Rb proteins are independent from the presence of human papillomavirus genes in oral squamous cell carcinoma.

BACKGROUND: The overexpression of p16(INK4A) and suppression of p53 and Rb proteins are key features of oncogenic transformation by human papillomaviruses (HPV) in anogenital cancers. HPV genomes are often detected in cancers of the oral cavity, but it is unclear whether HPV has a specific oncogenic role there. OBJECTIVES: The objectives of the study were to investigate the expression of p53, Rb, and p16(INK4A) proteins and identify HPV infection and viral integration into the host genome. METHODS: Seventy-nine cases of oral squamous cell carcinoma (OSCC) were studied by immunohistochemistry. Polymerase chain reaction (PCR) was performed to identify HPV DNA from the samples. The results were correlated with clinical data. RESULTS: Thirty-three cases were HPV positive for high-risk HPV (HR-HPV) types, of which 27 harbored HPV16. In 25 of 27 HPV16-positive tumors, the HPV16 genome was fully integrated into the host genome, as evidenced by the lack of PCR-amplifiable E2 gene sequences. Forty-five patients were p53 overexpressing, 20 with HR-HPV-positive and 25 with HR-HPV-negative tumors. p16(INK4A) protein was overexpressed in 4 of 31 HR-HPV-positive and 9of 45 HR-HPV-negative cases. Twenty-six of 32 HR-HPV-positive and 37 of 44 HR-HPV-negative samples exhibited pRb nuclear staining. These differences between HR-HPV-positive and -negative tumors were not statistically significant. No correlation was found between these biological factors and tumor location, stage, differentiation grade, or alcohol or tobacco abuse. CONCLUSIONS: A tumor immunophenotype, similar to HPV-related anogenital cancers, is not present in OSCC and highly oncogenic HPV types are therefore unlikely to be specific or independent risk factors for oral cancer.