Cyclin D1-negative Tumors Research Articles

Use of cyclin D1 in conjunction with human papillomavirus status to predict outcome in oropharyngeal cancer

There is increasing use of multiple molecular markers to predict prognosis in human cancer. Our aim was to examine the prognostic significance of cyclin D1 and retinoblastoma (pRb) expression in association with human papillomavirus (HPV) status in oropharyngeal squamous cell carcinoma. Clinical records and specimens of 226 patients with follow-up from 1 to 235 months postdiagnosis were retrieved. Tumor HPV status was determined by HPV E6-targeted multiplex real-time PCR/p16 semiquantitative immunohistochemistry and cyclin D1 and pRb expression by semiquantitative immunohistochemistry. Determinants of recurrence and mortality hazards were modeled using Cox regression with censoring at dates of last follow-up. The HPV-positivity rate was 37% (91% type 16). HPV was a predictor of recurrence, an event (recurrence or death) and death after adjustment for clinicopathological variables. There were inverse relationships between HPV status and cyclin D1 and pRb. On univariate analysis, cyclin D1 predicted locoregional recurrence, event and death and pRb predicted event and death. Within the HPV-positive group, after adjusting for clinicopathological factors, patients with cyclin D1-positive cancers had up to a eightfold increased risk of poor outcome relative to those with cyclin D1-negative tumors. However, within the HPV-negative group, there was only a very small adjusted increased risk. A combination of pRb and HPV did not provide additional prognostic information. Our data provide the first evidence that a combination of HPV and cyclin D1 provides more prognostic information in oropharyngeal cancer than HPV alone. If findings are confirmed, treatment based on HPV and cyclin D1 may improve outcomes.

Wnt/beta-Catenin pathway in human glioma: expression pattern and clinical/prognostic correlations

Gliomas are the most common primary intracranial tumors. Understanding the molecular basis of gliomas' progression is required to develop more effective therapies. The Wnt/β-catenin signaling cascade is an important signal transduction pathway in human cancers. Although, overactivation of this pathway is a hallmark of several forms of cancer, little is known about its role in human gliomas. Here, we aimed to determine the clinical significance of Wnt/β-catenin pathway components in gliomas. Immunohistochemical staining was performed to detect the expression patterns of Wnt1, β-catenin and Cyclin D1 in the biopsies from 96 patients with primary gliomas. Kaplan-Meier survival and Cox regression analyses were performed to evaluate the prognosis of patients. Cytoplasmic staining pattern of Wnt1, membranous, cytoplasmic and nuclear accumulation of β-catenin, and nuclear localization of Cyclin D1 were demonstrated by immunohistochemical staining. The Wnt1 expression significantly correlated with the expression of Cyclin D1 (P<0.0001). The ratio of tumors with a cytoplasmic-nuclear pattern or a cytoplasmic pattern of β-catenin was significantly higher in Wnt1-positive (P<0.01) and Cyclin D1-positive (P<0.01) tumors than in Wnt1-negative and Cyclin D1-negative tumors, respectively. The protein expression levels of Wnt1, β-catenin and Cyclin D1 were all positively correlated with the Karnofsky performance scale (KPS) score and World Health Organization (WHO) grades of patients with gliomas. Furthermore, Wnt1, cytoplasmic-nuclear β-catenin and Cyclin D1 status were all the independent prognostic factors for glioma patients (P=0.01, 0.007 and 0.005, respectively). These results provide convincing evidence that the Wnt/β-catenin pathway correlated closely with the progression of gliomas and might be a novel prognostic marker for this neoplasm.

Cyclin D1 Expression in Breast Cancer Patients Receiving Adjuvant Tamoxifen-Based Therapy

The objective of our study was to determine the clinical relevance of cyclin D1 expression in hormone receptor-positive breast cancer patients who were treated with tamoxifen-based therapy. We assessed expression of cyclin D1 in surgical specimens of breast carcinoma by means of immunohistochemistry. Patients had been enrolled in either Austrian Breast and Colorectal Cancer Study Group (ABCSG) Trial 05 or ABCSG Trial 06 and received tamoxifen as part of their adjuvant treatment. Overall survival and relapse-free survival were analyzed with Cox models adjusted for clinical and pathologic factors. Cyclin D1 was expressed in 140 of 253 (55%) tumors of ABCSG Trial 05 and in 569 of 948 (60%) tumors of ABCSG Trial 06. Expression of cyclin D1 was associated with poor outcome in both cohorts. Overall survival was significantly shorter in patients with cyclin D1-positive tumors compared with patients with cyclin D1-negative tumors [adjusted hazard ratio (HR) for death (ABCSG Trial 05), 2.47; 95% confidence interval (95% CI), 1.08-5.63; P = 0.03; adjusted HR for death (ABCSG Trial 06), 1.78; 95% CI, 1.36-2.34; P < 0.0001]. Relapse-free survival was also shorter in patients with cyclin D1-positive tumors than in patients with cyclin D1-negative tumors [adjusted HR for relapse (ABCSG Trial 05), 2.73; 95% CI, 1.50-4.96; P = 0.001; adjusted HR for relapse (ABCSG Trial 06), 1.52; 95% CI, 1.14-2.04; P = 0.005]. Cyclin D1 expression is an independent poor prognostic factor in women with early-stage, hormone receptor-positive breast cancer who received adjuvant tamoxifen-based therapy.

Cyclin D1 expression in breast cancer patients and tamoxifen therapy

523 Background: Adjuvant tamoxifen therapy improves survival of hormone receptor-positive breast cancer patients, but there is a subset of patients who fails to respond or develops resistance to tamoxifen. Cyclin D1 plays a central role in cell cycle regulation, directly affects the estrogen receptor and may predict outcome of tamoxifen therapy. Methods: We determined expression of cyclin D1 in formalin-fixed, paraffin-embedded surgical breast cancer specimens by means of immunohistochemistry. The patients had been enrolled in the Austrian Breast and Colorectal Cancer Study Group (ABCSG) Trial 5 (test set) or ABCSG Trial 6 (validation set). Comparison of cyclin D1 expression with clinical parameters and survival of the patients was performed with cyclin D1 as a continuous variable and as a dichotomized variable. The median value of the percentage of cyclin D1-positive tumor cells was a priori chosen as cut-off point to classify cyclin D1- positive and cyclin D1-negative tumors. The cutpoint was established in the test set and validated in the validation set. Overall survival and relapse-free survival were analyzed with Cox models adjusted for clinical and pathological factors. Results: Cyclin D1 was positive (&gt;10% cyclin D1-positive tumor cells) in 140 of 253 (55%) tumors of the test set and in 569 of 948 (60%) tumors of the validation set. A benefit from adjuvant endocrine therapy was associated with absence of cyclin D1 in both cohorts. Overall survival was significantly shorter in patients with cyclin D1-positive tumors as compared to patients with cyclin D1-negative tumors (adjusted hazard ratio for death [test set], 2.47, 95% confidence interval [1.08–5.63] P=0.03; adjusted hazard ratio for death [validation set], 1.91, [1.45–2.51] P&lt;0.001). Relapse-free survival was also shorter in patients with cyclin D1-positive tumors than in patients with cyclin D1-negative tumors (adjusted hazard ratio for recurrence or death [test set], 2.73, [1.50–4.96] P=0.001; adjusted hazard ratio for recurrence or death [validation set], 1.60, [1.19–2.16] P=0.002). Conclusions: Women with early-stage, hormone receptor-positive breast cancer and cyclin D1-negative tumors benefit more from adjuvant tamoxifen therapy than women with cyclin D1-positive tumors. No significant financial relationships to disclose.

Prognostic significance of cyclin D1 and p16 in patients with intermediate‐risk head and neck squamous cell carcinoma treated with docetaxel and concurrent radiotherapy

The current study aimed to evaluate the significance of the cell-cycle-control proteins cyclin D1 and p16 as prognostic markers in head and neck squamous cell carcinoma (HNSCC) patients treated with docetaxel and radiotherapy. Cyclin D1 and/or p16 protein expression was retrospectively evaluated by immunohistochemistry in 53 patients with stage T1-3N0-2M0 (except T1N0 glottis) HNSCC who were treated with 10 mg/m(2)/week docetaxel 4 to 6 times and received concurrent chemoradiotherapy. Kaplan-Meier univariate analysis revealed that patients with cyclin D1-positive tumors or p16-negative tumors had a worse prognosis compared with those with cyclin D1-negative tumors or p16-positive tumors (p = .0004 and p = .025, respectively). The prognostic significance of cyclin D1 expression, not p16 expression, was confirmed using a proportional hazard regression model. An assessment of cyclin D1 and p16 levels might be of clinical use in defining subgroups of patients with poor prognosis.

Prognostic significance of cyclin D1 expression in resected stage I, II non-small cell lung cancer in Arabs

The aim of this study was to evaluate the prognostic significance of cyclin D1 expression in primary, resected stage I and II non-small cell lung cancer (NSCLC) in Arabs. A longitudinal cohort study of 98 consecutive patients with resected stage I and II NSCLC were evaluated immunohistochemically for the expression of cyclin D1 and determined its prognostic significance by comparison with follow-up data from January 1994 to December 1999. This study included 76 male and 22 female patients. They represented 31 stage I NSCLC and 67 stage II tumors. Expression of cyclin D1 was detected immunohistochemically in 48 (49%) of the 98 NSCLC. Cyclin D1 expression had significantly higher positive results in patients with chronic obstructive pulmonary disease (P=0.001), poorly differentiated carcinoma (P=0.001), presence of vascular invasion (P=0.003), and visceral pleural invasion (P=0.005). Patients with cyclin D1-positive tumors had shorter survival than those with cyclin D1-negative tumors (5-year survival rates, 48% and 74%, respectively; P=0.006 by the log-rank test). In conclusion, a higher percentage of NSCLC with visceral pleural invasion, vascular invasion, poor differentiation of the tumor, patients with chronic obstructive pulmonary disease have positive cyclin D1 expression than other lung tumors.

Prognostic significance of cyclin D1 overexpression in patients (pts) with primary breast cancer (BC)

9714 Background: Cyclin D1, a key cell cycle regulator, is overexpressed in approximately 50% of BCs. Previous studies yielded conflicting results on the prognostic importance of cyclin D1 in BC. In this study, we evaluated overexpression of cyclin D1 in primary BC to determine any relation to clinicopathologic factors and its impact on survival. Methods: Formalin fixed tissue samples from 119 pts diagnosed with primary BC in 1996 and 1997 were analyzed for cyclin D1 expression by immunohistochemistry (IHC). A scoring system based on intensity and proportion of staining cells was used to judge cyclin D1 overexpression. ER, PR, Her2-Neu, p53 and Ki-67 expression by IHC and nuclear grade (NG) and histologic grade were determined at the time of diagnosis. OS and DFS curves were estimated by the method of Kaplan-Meier using log rank statistics. Clinicopathologic variables were included in Cox proportional hazards model to determine significance. Results: Cyclin D1 overexpression was observed in 68 of the 119 pts (57%). There was a positive correlation between cyclin D1 overexpression and ER expression (p=0.005) and an inverse correlation between cyclin D1 overexpression and both TNM stage and tumor size (p=0.009 and 0.042). Median follow up was 57 months. In univariate analysis, younger age, nodal involvement and higher NG were associated with poorer DFS (p=0.016, 0.001 and 0.026) and OS (p<0.001, p=0.016 and 0.036). Cyclin D1 had no influence on DFS or OS regardless of the cut point used to define overexpression. Among the ER positive subgroup however, there was a trend for improved OS in pts with cyclin D1 negative tumors (p=0.08). In contrast, cyclin D1 expression did not influence OS in ER negative subset. When ER positive pts with ductal carcinoma histology were considered, cyclin D1 negativity was associated with a better prognosis (p=0.053). In multivariate analyses of DFS and OS, only the nodal status (p=0.031 and 0.023) and NG (p=0.029 and 0.039) remained statistically significant. Conclusions: Cyclin D1 overexpression is not an independent predictor of prognosis in BC. In ER positive subgroup however, there is a trend for improved survival in cyclin D1 negative pts. No significant financial relationships to disclose.

The importance of cyclin D1 and Ki67 expression on the biological behavior of pancreatic adenocarcinomas

The aim of this study was to evaluate the role of cyclin D1 and Ki67 proteins involved in cell-cycle control as a prognostic factor in pancreatic carcinomas. We examined formalin-fixed, paraffin-embedded material from 59 pancreatic adenocarcinomas, for which appropriate clinical and prognostic data were available. The standard streptavidin–biotin immunoperoxidase method was used for immunostaining with cyclin D1 and Ki67. The extent of positive nuclear and cytoplasmic cyclin D1 staining was graded semiquantitatively. Ki67 reactivity was quantified and expressed as the percentage of stained nuclei. Staining with cyclin D1 and Ki67 was compared with histopathological prognostic features, and their relation with survival was also tested statistically. Patients whose tumors were cyclin D1-positive showed perineural invasion significantly more frequently than did patients with cyclin D1-negative tumors at the immunohistochemical level. In addition, tumors with lymphatic vessel invasion and without showed a significant difference in terms of cytoplasmic cyclin D1 staining. Ki67 indices were statistically different in stage groups. There was a significant and direct correlation between Ki67 index and nuclear cyclin D1 staining scores. No relation with survival was found. Our results suggest that cell-cycle proteins do not directly affect the prognosis of patients with pancreatic adenocarcinoma. Conversely, cyclin D1-positive tumors tend to have perineural invasion more frequently. In addition, lymph vessel invasion is another factor related to cyclin D1 reactivity of the cells. Ki67 indices differ statistically in stage groups.

Cyclin D1 Overexpression versus Response to Induction Chemotherapy in Squamous Cell Carcinoma of the Head and Neck?Preliminary Report

The aim of this study was to investigate whether there is an association between overexpression of cyclin D1 and response to therapy. Immunohistochemical overexpression of cyclin D1 was determined in paraffin-embedded specimens from diagnostic biopsies of 89 primary cases of squamous cell carcinoma of the head and neck (SCCHN), using a polyclonal antiserum. The tumor response rates were estimated after curative treatment (i.e. surgery and/or radiotherapy and/or chemotherapy). Patients whose tumors were overexpressing cyclin D1 showed complete or partial response to neoadjuvant chemotherapy with cisplatin/5-FU. In addition, a majority of cyclin D1 negative tumors did not respond at all to this treatment (p = 0.02, Fisher's exact test). This study indicates that immunohistochemical assessment of cyclin D1 expression in SCCHN could be a new predictive marker to select a subgroup of patients that will benefit from neoadjuvant chemotherapy.

Cyclin D1 expression is useful as a prognostic indicator for advanced esophageal carcinomas, but not for superficial tumors.

The purpose of the present study was to define the overexpression of cyclin D1 in superficial and advanced esophageal carcinomas and to investigate whether the expression of this molecule indicates a poor prognosis. This study included 41 patients with superficial esophageal carcinomas (Tis and T1) and 48 patients with advanced esophageal carcinomas (T2, T3, and T4). The expression of cyclin D1 in surgically resected specimens was evaluated immunohistochemically with a monoclonal antibody. Positive immunoreactivity was found in 31 of 89 cases (35%). Overexpression of cyclin D1 did not correlate with TNM classification or histologic type. Of the 48 patients with advanced esophageal carcinomas, 32 patients with cyclin D1-negative tumors survived longer than did 16 patients with cyclin D1-positive tumors (P = 0.0017). In contrast, we observed no survival difference between patients with cyclin D1-positive and -negative superficial esophageal carcinoma. These results suggest that cyclin D1 indicates a poor prognosis in cases of advanced esophageal carcinoma but not in cases of superficial esophageal carcinoma.

Expression of cyclin D1 is correlated with poor prognosis in patients with areca quid chewing-related oral squamous cell carcinomas in Taiwan.

Abnormal expression of cell cycle regulatory proteins, particularly cyclin D1, has been implicated in the pathogenesis of several types of cancer. We have examined the expression of cyclin D1 in histological sections of oral squamous cell carcinomas (SCCs) using anti-cyclin D1 antibodies with an immunoperoxidase technique. Cyclin D1 nuclear staining was observed in 73 of 88 (83%) cases of oral SCC. In 54 of these 73 (74%) cases, positive cyclin D1 staining was also found in the normal appearing epithelium immediately adjacent to the cyclin D1-positive SCCs. No significant correlation was found between the expression of cyclin D1 and the patients' age, sex, oral habits, cancer location and STNM status. The Kaplan-Meier analysis showed that patients with tumors containing more than 10% cyclin D1-positive cells had significantly shorter overall survival than those with tumors containing less than 10% cyclin D1-positive cells or with cyclin D1-negative tumors (P<0.05). Patients with positive lymph node status also had significantly shorter overall survival (P<0.01). These results indicate that cyclin D1 may play an important role in the genesis of oral SCC and may serve as an adjuvant marker of worse prognosis in patients with oral SCCs in Taiwan.

Overexpression of cyclin D1 and p53 associated with disease recurrence in colorectal adenocarcinoma.

Multiple genetic changes occur during the evolution of normal cells into cancer cells. It has been reported that both cyclin D1 and p53 genes play major roles in oncogenesis and/or cell cycle control in various cancers. In this study, we examined the overexpression of cyclin D1 and p53 by the immunohistochemical method and investigated the correlation between expression of these antigen and prognosis in patients with colorectal adenocarcinoma. Disease-free survival was significantly lower in the patients with cyclin D1-strongly positive tumors than in those with cyclin D1-negative tumors. Similarly, disease-free survival of the patients with p53-strongly positive tumors was significantly lower than that of those with p53-negative tumors. Moreover, multivariate analysis indicated that both cyclin D1 and p53 overexpression are independent prognostic factors in patients with colorectal adenocarcinoma. In conclusion, both cyclin D1 and p53 overexpression may be useful predictors of disease recurrence in patients with colorectal adenocarcinoma.

Overexpressed cyclin D1 protein related to p53 and retinoblastoma proteins in transitional cell carcinoma of the urinary tract

We employed immunohistochemical methods to investigate the overexpression of cyclin D1 protein as well as p53 protein and retinoblastoma protein (pRb) in 57 patients with transitional cell carcinoma (TCC) of the urinary tract, in order to determine their frequency and significance, and also association with the clinicopathological behavior of this tumor. Nuclear immunostaining with cyclin D1 antibody revealed 7 homogeneously stained tumors (12.3%), including 4 heterogeneous pRb expressions, of which only one was also positive for p53. Positive immunostaining with each antibody to p53 or pRb was obtained in 15 (26.3%) and 49 (86%) tumors, respectively. The 49 pRb-positive tumors included 6 homogeneous and 43 heterogeneous nuclear immunostaining patterns. The remaining pRb-negative 8 tumors included 5 cyclin D1-negative tumors. The prevalence of p53 staining was higher in high-grade (P<0.05) and invasive types of tumors (P<0.05) than the other types studied. On the other hand, no significant relationship was found between cyclin D1- or pRb-immunoreactivity and clinicopathological factors. These findings provide evidence for infrequent cyclin D1 protein overexpression in TCC independently associated with p53 protein and/or pRb overexpression.

Cyclin D1 overexpression in primary hypopharyngeal carcinomas.

Hypopharyngeal squamous cell carcinomas (HPCS) are associated with an extremely poor prognosis. Generally, conventional clinicopathologic factors have only limited value as prognostic factors for this malignancy. It is therefore clinically important to identify new prognostic factors that accurately reflect the biologic aggressiveness of this malignancy. The amplification and overexpression of the cyclin D1 protooncogene have been reported in a variety of malignancies, and are thought to be related to tumor progression. Based on this phenomenon, the authors immunohistochemically evaluated overexpression of the cyclin D1 gene in 42 cases of primary HPCS. In addition, the immunohistochemical staining of the proliferation marker MIB-1 (Ki-67 antibody) was also performed. Formalin fixed, paraffin embedded biopsy specimens obtained prior to treatment were examined. Cyclin D1 and Ki-67 were detected using monoclonal antibodies by means of the streptavidin-biotin method. The relationship between cyclin D1 overexpression and the stage, histologic grade, presence of lymph node metastases, proliferation index, and survival was then statistically analyzed. The correlation between the proliferation index, other clinicopathologic factors, and survival was also evaluated. Twenty-three (54.8%) HPCS specimens showed a 20% or greater immunoreactivity for cyclin D1. Cyclin D1 overexpression was related to cervical lymph node metastases (P = 0.037) but not to clinical stage, histologic grade, or the proliferation index. Cyclin D1 negative tumors were associated with a significantly better prognosis (P = 0.023), particularly in patients who underwent multimodality treatment. Finally, the MIB-1 labeling index showed no correlation with either the clinicopathologic parameters or overall survival. Based on these findings, cyclin D1 immunohistochemical staining is considered to be useful, not only as a prognostic factor for HPCS, but also as a means of determining the optimum treatment for each individual patient. Conversely, the MIB-1 labeling index appears to have no clinical significance in HPCS.