Immunofluorescence analysis of DNA damage response protein p53-binding protein 1 in a case of uterine dedifferentiated leiomyosarcoma arising from leiomyoma

AimsGenomic instability has been indicated during the dedifferentiation process from leiomyoma (LM) to leiomyosarcoma (LMS). Previously, we have described that nuclear expression pattern of DNA damage response protein p53-binding protein 1 (53BP1), detected by immunofluorescence, reflects the magnitude of genomic instability during malignancy. Here, we present a case of LMS arising from LM with molecular analysis of 53BP1, which showed transitional magnitude of DNA damage response within a tumor.Methods and resultsA fifty-year-old female with abdominal mass underwent hysterectomy. Histologically, the tumor consisted of LMS with highly atypical multinucleated giant cells as well as an LM component with transitional atypical spindle cells in the border area. LMS showed diffuse nuclear staining of 53BP1 expression, which has been previously described as high DNA damage response pattern. In contrast, the LM component lacked 53BP1 immunoreactivity and focal expression was observed in transitional lesion. Furthermore, double-labelled immunofluorescence revealed co-localization of 53BP1 with p53 and Ki-67 in the LMS component, which indicated abnormal DNA damage response in proliferative state.ConclusionsThis study revealed that diffuse-type 53BP1 expression may be beneficial to estimate genomic instability during dedifferentiation from LM to DLMS.     

软组织平滑肌肉瘤中肿瘤抑制基因p15,p16的表达

目的：探讨p15,p16蛋白在软组织平滑肌肉瘤（LMS）及平滑肌瘤（LM）的表达情况及其意义,方法：采用免疫组织化学方法检测38例LMS及20例LM标本的p15,p16蛋白表达情况,结果：在LMS,p15,p16的阴性率分别为18．42％及42．1％,二者差异有显著性。p15在LM的阴性率为60％,高于LMS。38例LMSp15和p16总异常率为52．6％,其中Ⅰ级LMS异常率显著高于Ⅱ、Ⅲ级。随访15例LMS,复发死亡组p15和p16异常率达75％,高于无复发组。结论：在LMSp16表达缺失比p15缺失更为常见,p15的缺失在LM在比在LMS常见,LMS的发生发展及预后p15及p16共同失活有关。     

Prognostic factors in malignant mixed tumors of the salivary gland: correlation of immunohistochemical markers with histologic classification

Malignant mixed tumor of salivary glands is a rare tumor whose variable behavior and prognosis are related for the most part to the clinical stage and histologic grade of the carcinomatous component. The purpose of this study is to predict prognosis by comparing the histologic grading and subclassification of the carcinomatous component with the immunohistochemical reactivity for E-cadherin, P53 mutation protein, and cellular proliferation (Ki67). Stains were performed on formalin-fixed paraffin-embedded tissue sections from 18 cases of malignant mixed tumor. Clinical follow-up was obtained for each patient. Regional lymph node and distant organ metastases were the criteria for poor prognosis. Of seven cases with lymph nodes metastasis, five were high-grade tumors (with one subsequent death from brain metastasis) and two were low-grade. Of the eight high-grade tumors, positivity for Ki67, p53, and E-cadherin were noted in six, four, and two cases, respectively. In contrast, of the 10 low-grade tumors, two stained with Ki67, five with p53, and none with E-cadherin. Most notably, all seven metastatic cases (as opposed to only one of 11 nonmetastatic tumors) had Ki67 reactivity of more than 10%. We conclude that malignant mixed tumor represents a spectrum of malignancies in which the clinical behavior is closely related to the carcinomatous element. In addition to histologic grading, Ki67 is a useful prognostic marker in the evaluation of malignant mixed tumor while p53 and E-cadherin appear to be of limited value.     

Diagnostic utility of cell cycle and apoptosis regulatory proteins in verrucous squamous carcinoma.

A major problem in the diagnosis of verrucous squamous cell carcinoma is the lack of readily reproducible objective criteria for distinguishing this malignant lesion from reactive epithelial hyperplasia. Both lesions are characterized by thickened (well-differentiated) squamous epithelium without cellular atypia and subjacent stroma densely infiltrated by lymphocytes and plasma cells. This study was carried out to evaluate the use of cell cycle and apoptosis-related regulatory proteins in the diagnosis of verrucous carcinoma. The study materials consisted of representative formalin-fixed and paraffin-embedded tissue blocks from 19 cases of verrucous carcinoma, 18 classic squamous cell carcinoma, and 14 squamous epithelial hyperplasia (acanthosis). The immunohistochemical expression of the following of cell cycle and apoptosis-related regulatory proteins was evaluated using avidin-biotin complex detection technique: p16, p21, p53, Ki67, and retinoblastoma gene product (RBGP) (also known as retinoblastoma protein [pRb]). Expression of Ki67 was detected only in the single basal layer of the epithelium in all 14 cases of acanthosis. In verrucous carcinoma, Ki67 was detected in basal and suprabasal cells in the lower third of the neoplastic epithelium in 19 of 19 cases (100%). In neoplastic squamous epithelium with frankly invasive squamous cell carcinoma, Ki67 was diffusely expressed throughout the entire thickness of the epithelium as well as in the underlying invasive tumor nests. The pattern of p53 expression was similar to that of Ki67 in all the experimental groups, with a Pearson correlation coefficient of 0.98. In addition, immunohistochemical expression of p53 in the hyperplastic squamous epithelium was very weak, in contrast to the more intense immunoreactivity observed in verrucous carcinoma and classic squamous cell carcinoma. There was an overlapping in the expression of p16, p21, and RGBP in all the experimental groups, being present in more than half the thickness of the epithelium in 50% to 100% cases in each study group. We therefore conclude that the pattern of Ki67 and p53 expression in verrucous carcinoma is readily reproducible and distinctly different from that observed in epithelial hyperplasia and that seen in invasive squamous cell carcinoma. Thus Ki67, and p53 immunostains are reliable adjuncts that may be helpful in resolving diagnostic problems associated with verrucous carcinoma.     

Malignant transformation of mature cystic teratoma to squamous cell carcinoma involves altered expression of p53‐ and p16/Rb‐dependent cell cycle regulator proteins

Ovarian mature cystic teratomas (MCT) uncommonly undergo malignant transformation to squamous cell carcinoma (SCC). While alterations in the p53 tumor suppressor gene and protein have been shown, few studies have analyzed other molecular changes leading to this malignant conversion. The purpose of the present study was to investigate 21 samples of SCC arising in MCT for altered expression in known p53‐ and p16/Rb‐dependent cell cycle regulatory proteins, and the association between their expression and cellular proliferation and histological features. Overexpression of the p53 protein was observed in 14 SCC (67%), while four (19%) had point mutations in the p53 gene. Reduced expression of the p16 protein was observed in 18 SCC (86%), while p16 gene alterations (hypermethylation (29%) and point mutation (33%)) were found in 11 (52%). Furthermore, a statistically significant correlation was observed between p53 and Rb overexpression (P = 0.0010), and the overexpression of both p53 and Rb was respectively significantly correlated with increased cellular proliferation. The results indicate that alterations in both the p53 and p16‐Rb pathways are associated with SCC arising in MCT.     

Leptomeningeal melanomatosis associated with neurocutaneous melanosis: An autopsy case report

An autopsy case of leptomeningeal melanomatosis associated with neurocutaneous melanosis (NCM) involving a 44‐year‐old male is reported. The autopsy showed that the leptomeningeal surface of the brain and the spinal cord were covered with a diffuse black lesion. A histological examination detected diffusely distributed, proliferating, melanin‐containing cells and demonstrated that the lesion consisted of three different components; i.e. regions of melanomatosis, melanocytosis, and melanocyte hyperplasia. In the leptomeningeal melanomatosis component, tumor cells with pleomorphic nuclei and prominent nucleoli had infiltrated into the cerebral parenchyma via Virchow–Robin spaces. The Ki‐67 labeling index and the nuclear accumulation of p53 and p16 protein were immunohistochemically examined in each component. The Ki‐67 labeling indices of the melanomatosis, melanocytosis, and melanocyte hyperplasia components were 8.7%, 0.8%, and 0%, respectively. Immunostaining of nuclear p16 produced a negative result in the melanomatosis component, but positive results in the melanocytosis and melanocyte hyperplasia components, whereas nuclear p53 expression was not detected in any of the components. This case suggests that p16^INK4/CDKN2 may play a significant role in progression of leptomeningeal melanocytic neoplasms. We also reviewed previously reported cases of leptomeningeal neoplasms associated with NCM and discussed the relationship between the biological behavior and proliferative activity of such lesions.     

p53与Ki67在判断困难的子宫平滑肌肿瘤诊断中的价值

目的：探讨免疫组化方法在疑难的子宫平滑肌肿瘤诊断中的价值。方法：对56例判断困难的子宫平滑肌肿瘤进行光镜形态以及p53和Ki67标记的观察。结果：46例平滑肌瘤主要表现单项形态异常，p53阳性标记占8.7%，Ki67阳性细胞数平均65.87。7例平滑肌肉瘤或恶性倾向未定的平滑肌肿瘤主要表现多项形态异常，p53阳性标记占85%,Ki67阳性细胞数平均683.43。结论：p53与Ki67标记可以共同作为有价值的客观指标，为子宫平滑肌肿瘤的良、恶性判断提供辅助诊断依据。     

Goblet cell status in idiopathic ulcerative colitis--implication in surveillance program

Idiopathic ulcerative colitis (IUC) patients have higher incidence of dysplasia and malignancy. Close follow-up with biopsy at regular interval is mandatory. The study was done to correlate incidence of atypical epithelium, goblet cell hyperplasia (GCH) and disease duration (DD) with Ki67, AgNOR and p53 expression in IUC with disease for 5 or more years. Ki67 and AgNOR are good indicators of cellular proliferation and p53 tumour suppressor protein is a marker for neoplastic cell. Of 130 cases studied, 40 cases showed atypical epithelium and were selected for further study. DD in these 40 cases ranged from 60 to 228 months. All had GCH and showed histological features of chronicity. Low-grade dysplasia (LGD) was seen in 15 cases, indefinite for dysplasia (ID) in 8 and inflammatory atypia in 17 cases. Disease duration showed no influence in the type of atypical epithelium. A positive staining of lining epithelium by Ki67 and p53 was not restricted to dysplasia. LGD and ID showed stronger p53 nuclear staining. AgNOR appeared to be a more sensitive marker than Ki67. GCH showed a positive correlation with DD and AgNOR index. p53 expression correlated positively with goblet cell hyperplasia. Conclusion- goblet cell hyperplasia could indicate presence of epithelial cell dysplasia.     

Immunohistochemical and molecular analysis of p53, MDM2, proliferating cell nuclear antigen and Ki67 in benign and malignant peripheral nerve sheath tumours

A series of 26 malignant peripheral nerve sheath tumours (MPNST) and 24 benign peripheral nerve sheath tumours (BPNST) were analysed immunocytochemically for p53 expression and the cell proliferation markers proliferating cell nuclear antigen (PCNA) and Ki67 (with MIB1). In 23/26 MPNST, 5%–65% of the tumour cell nuclei were immunoreactive for Ki67 with MIB1 while none of the 24 BPNST had nuclear staining exceeding 5%. Greater than 50% nuclear PCNA staining was detected in 25/26 MPNST compared with 8/24 BPNST; 17/26 MPNST showed 5–100% nuclear staining for p53 (13/26>20%), whereas none of the BPNST had nuclear staining exceeding 1%. The Ki67, PCNA and p53 immunostaining results correlated significantly with benign versus malignant (P<0.001, P<0.001 and P<0.005, respectively) as well as mitotic rate (P<0.001, P<0.05 and P<0.05). Ki67 immunostaining results correlated significantly with PCNA and p53, as did p53 with Ki67 and PCNA (P<0.001 in both). Stepwise (logistic regression forward) multivariate analysis of the variable, benign versus malignant, revealed the strongest correlations with PCNA (P=0.007) and Ki67 (P=0.021). Direct confirmation of the presence of p53 protein was obtained by western blot analysis of 3 MPNST and 5 BPNST. Two MPNST, showing 90% and 30% immunoreactivity, were positive for p53, while one MPNST with 5% immunoreactivity and all 5 BPNST were negative. Southern blot analysis performed on the two MPNST with high p53 protein levels revealed no amplification of the MDM2 gene, suggesting that high p53 levels in MPNST are likely to be due to mutation. The results also indicate that PCNA and Ki67 are potentially useful in distinguishing BPNST from MPNST, particularly in problematic cases of cellular schwannoma versus MPNST. The detection of p53 in a large percentage of cells of a plexiform neurofibroma giving rise to MPNST and Ki67 in 5% and 25% of cells of two similar cases suggests that malignant transformation may be detected in some cases by p53 and proliferation markers prior to overt histological evidence of malignancy.     

HDM2 overexpression and focal loss of p14/ARF expression may deregulate the p53 tumour suppressor pathway in meningeal haemangiopericytomas. Study by double immunofluorescence and laser scanning confocal microscopy

AIMS: To investigate the p53 pathway in meningeal haemangiopericytomas (MHPCs), p14/ARF, p53 protein expression and two wild-type (wt) p53-induced proteins (HDM2 and p21/WAF1) were studied in 18 MHPCs, 11 primary, four of them recurrent on one, one, two and four occasions. METHODS: Immunohistochemical detection of p14/ARF, p53, p21/WAF1, HDM2 and Ki67 proliferative index (PI) protein expression. RESULTS: Ki67 index was > 5% in eight out 18 cases (44.4%). The PI in recurrent cases increased with neoplastic progression. Simultaneous p53 and wt p53 transactivated gene (p21/WAF, HDM2) expression occurred in all cases. This argues against p53 mutation. HDM2 overexpression was observed in 10 cases (55.5%). Double-immunofluorescence staining and laser scanning confocal microscopy (LSCM) displayed HDM2 and p53 colocalization. This strongly suggests that HDM2 binds and inactivates p53 that could be pathogenic for MHPCs, by a different mechanism than point mutation. p14/ARF expression > 5% was observed in 12 cases (66.6%). A normal (diffuse) pattern of expression was seen in 13 cases (72.2%). Focal loss of expression was observed in five patients (27.7%): three primary cases and two recurrences. Therefore, p14/ARF down-regulation may also contribute to the development of MHPC. CONCLUSION: HDM2 overexpression, sometimes combined with focal loss of p14/ARF expression, may play a pathogenic role in MHPCs.     

Malignant peripheral nerve sheath tumours: high Ki67 labelling index is the significant prognostic indicator

AIMS: We investigated p53, Ki67, MDM2, and p21WAF1/CIP1 in order to evaluate its relationship with prognosis in malignant peripheral nerve sheath tumours (MPNST). METHODS AND RESULTS: In 49 cases of MPNSTs, the immunohistochemical studies of Ki67, p53, MDM2, p21WAF1/CIP1 and polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) with direct sequencing of p53 were performed with the formalin-fixed paraffin-embedded tissue. In 43 cases with survival data available, an evaluation of the prognostic significance of clinicopathological factors was also carried out. A high Ki67 labelling index (LI) (>25%) was correlated with a reduced survival rate in the 43 cases of MPNST (P=0.0106, log-rank test). Furthermore, there was a significant correlation between the Ki67 LI and the immunohistochemical expression of p53 or MDM2. In 17 MPNST cases, PCR amplification of exons 5 through 8 of the p53 gene was successful. One case showed a base change of codon 240 (AGT-->AGC), but translated amino acid (Ser) remained unchanged. Multivariate Cox analysis of our series showed that the association of von Recklinghausen's disease, tumour depth, and the presence of rhabdomyoblasts (malignant triton tumour) each had an independent negative impact on overall survival. CONCLUSION: High Ki67 LI (>25%) was of significant prognostic value in MPNST.     

A prognostic index in skin melanoma through the combination of matrix metalloproteinase-2, Ki67, and p53

The objective of this immunohistochemical study was to explore the roles of Ki67 and p53 in conjunction with matrix metalloproteinase-2 and matrix metalloproteinase-9, tissue inhibitors of metalloproteinase-1 and tissue inhibitors of metalloproteinase-2 in a series of 157 cases of skin melanomas. Elevated Ki67 expression and positive staining for p53 correlated to the propensity to metastasize (P = .016) and to declined disease-specific survival, as well as to shortened recurrence-free survival. In patients with a high immunoreaction for Ki67, the 10-year disease-specific survival was 39% compared with 73% in patients with a low Ki67 expression (P = .03). In cases with a positive p53 expression in melanoma cells, the 10-year disease-specific survival was 59% compared with 76% in patients with a negative immunoreaction for p53 (P = .005). Overexpression of the matrix metalloproteinase 2 protein in conjunction with overexpression of Ki67 characterized melanomas with high metastatic potential and was associated with declined survival with a 10-year disease-specific survival of 33% compared with 85% in the cases with low matrix metalloproteinase-2 and low Ki-67 levels (P = .002). Similarly, in cases with overexpression of matrix metalloproteinase-2 and a positive immunoreaction for p53, the 10-year disease-specific survival was only 42% compared with 80% in patients with matrix metalloproteinase-2 less than 20% and a negative immunostaining for p53 (P < .001). The presence of all 3 adverse prognostic factors was prognostically more significant than any marker alone with a 10-year survival of only 28%. This combination of determining matrix metalloproteinase 2, Ki67, and p53 immunoreactive proteins could be beneficial in the selection of high-risk melanoma patients for future adjuvant trials.     

Immunohistochemical assessment of Ki67 and p53 expression assists the diagnosis and grading of ulcerative colitis-related dysplasia

AIMS: To assess whether Ki67 and p53 immunostaining may assist the diagnosis and grading of ulcerative colitis-related dysplasia. METHODS AND RESULTS : Location of Ki67 staining and location and intensity of p53 staining were assessed in ulcerative colitis (UC) cases showing the features of high-grade dysplasia (HGD, n = 14), low-grade dysplasia (LGD, n = 22), 'indefinite for dysplasia' (n = 12), or regenerative atypia (RA, n = 22). Good intra- and inter-observer reproducibilities were demonstrated in the performance of these assessments. All the dysplasia cases showed extension of Ki67 staining above the basal third of the crypt. Moderate intensity p53 staining was seen in 10/22 RA cases, but strong intensity p53 staining was seen only in cases of dysplasia. All the cases of HGD showed extension of Ki67 and p53 staining above the basal two thirds of the crypt. CONCLUSIONS: Restriction of Ki67 staining to the basal third of the crypt appears to exclude a diagnosis of dysplasia whereas strong intensity p53 staining suggests a diagnosis of dysplasia. Restriction of Ki67 or p53 staining to the basal two-thirds of the crypt appears to exclude a diagnosis of HGD.     

Lack of histologically suspicious features,proliferative activity,and p53 expression suggests benign diagnosis in phaeochromocytomas

AIMS: The malignancy of phaeochromocytomas is difficult to predict. Traditionally, only a metastasized tumour is considered malignant. The aim of this study was to assess the histopathological and clinical features, as well as the proliferative activity, and to analyse p53 and p21 expression in 105 phaeochromocytomas. METHODS AND RESULTS: All malignant phaeochromocytomas (n = 8) showed at least one of the histologically suspicious features, i.e. over five mitoses/10 high-power fields, necrosis, capsular or vascular invasion. Malignant tumours were larger, but the age and gender of the patients were not significantly different. All benign (n = 33) and most of the borderline (18/21) adrenal phaeochromocytomas had less than 6% Ki67+ tumour cells, while most malignant tumours (6/7) expressed Ki67 in >6% of the cells. p53+ immunohistochemistry was found in two malignant tumours, while p21 expression did not correlate with malignancy. CONCLUSIONS: These data suggest that the lack of histologically suspicious features, low proliferative activity, smaller size, and negative p53 immunostaining point to a benign diagnosis in phaeochromocytomas.     

Expression of cell cycle-regulatory proteins, MIB-1, p16, p53, and p63, in squamous cell carcinoma of conjunctiva: not associated with human papillomavirus infection

Squamous cell carcinoma (SCC), the most common primary malignant tumor of the conjunctiva, has a variable clinical presentation and immunohistochemical profile. Abundant cell cycles exist, including MIB-1 (Ki67 antigen), p16, p53, and p63, within the conjunctiva SCC. This investigation first reports the expressions of cell cycle markers in SCC. A retrospective study was conducted between December 1976 and June 2004, comprising 13 consecutive patients with conjunctiva SCC who were treated with surgical excision. Detailed clinical parameters were also reviewed. Overexpression of MIB-1, p16, p53, and p63 genes were studied by immunohistochemistry. Genechip containing 39 subtypes was used to elucidate human papillomavirus (HPV). The study group contained 13 (100%) men, with a mean age of 68±18 years and follow-up period of 20±17 months. The sample included four (33%) SCC located in the left eye and two (17%) recurrent SCC. Overexpression of the p53 and p63 was considerably higher than that of the p16 (P<0.01). HPV DNA was not detected in any of the 13 cases. This work first examined the immunohistochemical overexpression of cell cycle (MIB-1, p16, p53, and p63) in SCC. This investigation then showed that the expression of cell cycles in SCC was associated with key tumor clinicopathological features. This approach can help distinguish the potential roles of cell cycle in the development of SCC.     

Enhanced cell kinetics, p53 accumulation and high p21WAF1 expression in chronic cholecystitis: comparison with background mucosa of gallbladder carcinomas

AIMS: Since neoplasia resulting from chronic inflammation has recently attracted increasing attention, we have investigated surgically removed gallbladders to examine the relationship between chronic cholecystitis and carcinogenesis. METHODS AND RESULTS: The mucosa of 108 cholecystectomy specimens without gallbladder cancer and 54 surgically resected gallbladder carcinomas were classified into three groups according to the degree of lymphocytic infiltration, and assessed immunohistochemically for Ki67, p53, p21WAF1 and apoptosis. In gallbladder mucosa without carcinoma, all four parameters tended to increase with the inflammation score (IS). Significantly positive correlations were revealed between Ki67 and p53, Ki67 and p21WAF1, and p53 and p21WAF1. However, in gallbladder carcinoma cases, values of p53 and p21WAF1 for background mucosa were elevated as compared to the mucosa of cholecystitis with low IS, but there was no correlation between their expression and IS, except for Ki67. CONCLUSIONS: Severe chronic cholecystitis is associated with acceleration of epithelial cell turnover, damaged cells being eliminated by apoptosis. The background mucosa of gallbladder carcinomas showed similar cell proliferative activity (Ki67) to that in cholecystitis, with no parallel changes of p53 and p21WAF1 expression, suggesting the possibility of unknown cofactors causing genomic damage.     

4种生物学因子对局部晚期乳腺癌新辅助化疗疗效评估的价值

目的 探讨p53、ki67、nm23、表皮生长因子受体(EGFR)在局部晚期乳腺癌新辅助化疗疗效评估中的价值。 方法 选取2013年1月1日~2018年3月31日我院收治的局部晚期乳腺癌患者98例，检测新辅助化疗前后p53、ki67、nm23、EGFR的变化及其与临床疗效的关系。 结果 化疗后的p53、ki67、EGFR阳性表达均较化疗前降低，nm23阳性表达均较化疗前升高，差异均有统计学意义（P<0.05）。p53阳性者的有效率低于阴性者，差异有统计学意义（P<0.05）；ki67、nm23阳性者的有效率均高于阴性者，差异均有统计学意义（P<0.05）；EGFR阳性者与阴性者的有效率差异无统计学意义（P>0.05）。 结论 新辅助化疗能够显著改善局部晚期乳腺癌患者生物学因子p53、ki67、EGFR、nm23的表达，同时p53、ki67、nm23对评估化疗疗效具有一定价值。     

Angiomyolipoma of the kidney: expanding disease spectrum demonstrated by 3 cases.

We report 3 recent cases of angiomyolipoma of the kidney. Although generally regarded as a benign neoplasm, angiomyolipoma rarely behaves in an aggressive manner, producing complicated clinical courses leading to metastasis and death. The presence of epithelioid elements within the tumor can result in difficulty differentiating benign from malignant angiomyolipoma and differentiating this tumor from renal adenocarcinoma. The presence of lymph node involvement can cause difficulty in differentiating multicentric disease in lymph nodes from metastasis to lymph nodes. The presence of cytologic abnormalities in the primary tumor can result in difficulty in differentiating atypia in benign angiomyolipoma from malignant sarcomatous transformation of a benign lesion. The 3 cases reported show many of these problems. Criteria for predicting malignancy in epithelioid tumors and sarcomatous transformation are not well recognized because of the rarity of this entity. The typical immunophenotype of all types of angiomyolipoma (cytokeratin-negative and melanomarkers-positive) is very useful in diagnosis but does not help in the differentiation from renal adenocarcinoma at frozen section. We report the empiric use of Ki67 and p53 in these cases as adjuncts to clinical and histologic assessment in predicting behavior. High Ki67 expression was a feature of malignant epithelioid angiomyolipoma. Low levels of p53 expression were seen in the angiomyolipoma with sarcomatous transformation. Benign angiomyolipomas were consistently negative for both Ki67 and p53.     

Comparative study of the expression of cellular cycle proteins in cervical intraepithelial lesions

Interaction of human papilloma virus oncoproteins E6 and E7 with cell cycle proteins leads to disturbances of the cell cycle mechanism and subsequent alteration in the expression of some proteins, such as p16^INK4a, cyclin D1, p53 and KI67. In this study, we compared alterations in the expression of these proteins during several stages of intra-epitelial cervical carcinogenesis. Accordingly, an immunohistochemical study was performed on 50 cervical biopsies, including negative cases and intraepithelial neoplasias. The expression patterns of these markers were correlated with the histopathological diagnosis and infection with HPV. The p16^INK4a, followed by Ki67, showed better correlation with cancer progression than p53 and cyclin D1, which recommends their use in the evaluation of cervical carcinogenesis. These monoclonal antibodies can be applied to cervical biopsy specimens to identify lesions transformed by oncogenic HPV, separating CIN 1 (p16^INK4a positive) and identifying high-grade lesions by an increase in the cellular proliferation index (Ki67). In this way, we propose immunomarkers that can be applied in clinical practice to separate patients who need a conservative therapeutic approach from those who require a more aggressive treatment.     

Evaluation of CINtec PLUS® testing as an adjunctive test in ASC‐US diagnosed SurePath® preparations

The CINtec PLUS® system is an immunohistochemical cocktail composed of antibodies against p16^INK4a (surrogate of HPV infection) and Ki‐67 (proliferation marker) meant to improve the sensitivity and specificity for detecting high‐grade dysplasia (HGD). In the presence of dysplasia, a red chromogen marks Ki‐67 expression in the nucleus and a brown chromogen marks cytoplasmic p16^INK4a expression. Only cells showing dual staining are interpreted as positive. This retrospective study examined the performance of CINtec PLUS testing when performed on ASC‐US diagnosed samples. Comparison was made to high‐risk HPV DNA test results and colposcopic biopsy results. Technical considerations in the interpretation of this immunohistochemical stain are additionally discussed. CINtec PLUS showed modest sensitivity (64%) and specificity (53%) in identifying the presence of HGD at surgical biopsy. Positive and negative predictive values for HGD were 28% and 83%, respectively. HR‐HPV DNA test yielded sensitivity of 100% and specificity of 21%. During interpretation, squamous metaplasia and endocervical cells were seen to show individual staining for p16^INK4a or Ki‐67. Individual staining, when present within three dimensional cellular groups common to SurePath® preparations, can be time‐intensive to interpret necessitating thoughtful examination at high power. The Pap test with HR‐HPV DNA testing is a highly sensitive test. A specific test is needed to prevent false positives and over treatment. The CINtec® system provides a modest increase in specificity beyond HR‐HPV DNA testing. Future study of its appropriateness and cost‐ffectiveness in a treatment algorithm are warranted. Diagn. Cytopathol. 2013. © 2011 Wiley Periodicals, Inc.