Prognostic significance of methylated RASSF1A and PITX2 genes in blood- and bone marrow plasma of breast cancer patients.

Free circulating DNA is increased in the serum/plasma of cancer patients, and methylation of certain genes has been found to be characteristic for malignancy. Therefore, we investigated the prognostic value of two promising genes, PITX2 and RASSF1A, in peripheral blood-plasma (PB-P) and bone marrow plasma (BM-P) of breast cancer patients. Peripheral blood and bone marrow samples from patients with primary breast cancer were prospectively collected during primary surgery at the Department of Obstetrics and Gynecology in Innsbruck (n = 428) from June 2000 to December 2006. The study has been approved by the ethical committee of the Medical University of Innsbruck. Methylation analysis was performed using MethyLight, a methylation-specific quantitative PCR-method. In univariate survival analysis, methylated PITX2 in PB-P was found to be a significant indicator for poor overall survival (OAS) and distant disease-free survival (DDFS) (P = 0.001 and P = 0.023). Methylated RASSF1A in PB-P was also an indicator for poor OAS and DDFS (P = 0.001 and P = 0.004). RASSF1A had also significant prognostic potential when determined in BM-P (P = 0.016). In multivariate survival analysis methylated PITX2 and RASSF1A in PB-P remained as therapy-independent prognostic factors for OAS (P = 0.021, P < 0.001). For DDFS only RASSF1A in PB-P showed prognostic significance (P = 0.002). Methylated RASSF1A and PITX2 in PB-P appear to have promising potential as prognostic markers in clinical use.

Association of elevated C-reactive protein levels with an impaired prognosis in patients with surgically treated endometrial cancer.

OBJECTIVE: To evaluate whether C-reactive protein (CRP) serum levels are associated with prognosis in surgically treated endometrial cancer. METHODS: In the present multicenter study, CRP serum levels were measured preoperatively in 403 surgically staged patients with endometrioid endometrial cancer. Results were correlated to clinical data. RESULTS: The mean (standard deviation) serum CRP level in patients with endometrial cancer was 1.0 (1.8) mg/dL. Serum CRP levels were associated with tumor stage (P=.01), but not with tumor grade (P=.8), lymph node involvement (P=.8), and age at diagnosis (P=.9). In a univariable survival analysis, serum CRP levels, tumor stage, tumor grade, and age at diagnosis were associated with disease-free and overall survival (all P <.001). In a multivariable Cox regression model, serum CRP levels (P=.001, P=.004), tumor stage (P <.001, P <.001), tumor grade (P=.02, P=.009), and age at diagnosis (P=.002, P=.001) were independent prognostic factors for disease-free and overall survival. CONCLUSION: Our results suggest that elevated serum CRP levels are associated with a less favorable prognosis in patients with surgically treated endometrial cancer. LEVEL OF EVIDENCE: II.

Disseminated tumor cells: are they ready for clinical use?

During the past three decades, efforts successfully established the presence of disseminated tumor cells (DTC) in bone marrow as a prognostic factor. These works were comprehensively evaluated in a pooled analysis that now permits to classify the prognostic significance of DTC as level I evidence. Intriguing molecular data suggest a role for tumor stem cells possibly responsible for the prognostic impact of DTC. In a typical clinical setting of the year 2007, DTC--irrespectively of the strong prognostic significance--would only have a convincing clinical application if DTC were a surrogate marker for treatment efficacy. Consequently, this important question is to be addressed in well-designed clinical trials.

Analysis of aberrant DNA methylation and human papillomavirus DNA in cervicovaginal specimens to detect invasive cervical cancer and its precursors.

PURPOSE: Cancer of the uterine cervix is an important cause of death in women worldwide. Pap smears as a tool for screening decreased the incidence and mortality of cervical cancer dramatically. This proof of principle study aimed to develop a potential tool for cervical screening using a test that can be applied by patients without visiting a physician and to increase the coverage rate, especially of the high-risk population with low socioeconomic status. EXPERIMENTAL DESIGN: Human papillomavirus (HPV) DNA testing and methylation analysis of DNA obtained from cervicovaginal specimens of 13, 31, and 11 patients with no dysplasia/low-grade squamous intraepithelial lesion (SIL), high-grade SIL, and invasive cervical cancer, respectively, collected on a tampon, was performed using PCR-based methods to detect invasive cervical cancer and study whether these changes are already present in the precursor lesions. RESULTS: High-risk HPV DNA was present in 68 and 82% of patients with high-grade SIL and invasive cervical cancer. DNA methylation of the 11 genes tested increased with severity of the cervical lesion. Unsupervised hierarchical cluster analysis using solely information on DNA methylation of the 11 genes was able to predict the presence of invasive cervical cancers: one of the two clusters formed contained 9 of 11 invasive cervical cancers, as well as two high-grade SILs. CONCLUSIONS: HPV DNA and DNA methylation analyzed in cervicovaginal specimens are able to predict invasive cervical cancers. To detect all high-grade SILs when applying this test, genes that become methylated earlier throughout cervical carcinogenesis have to be defined.

Methylated DNA collected by tampons--a new tool to detect endometrial cancer.

This proof of principle study aimed to define a new and simple strategy for detection of endometrial cancer using epigenetic markers. We investigated DNA isolated from vaginal secretion collected from tampon for aberrant methylation of five genes (CDH13, HSPA2, MLH1, RASSF1A, and SOCS2) using MethyLight in 15 patients with endometrial cancer and 109 patients without endometrial cancer. All endometrial cancer patients revealed three or more methylated genes, whereas 91% (99 of 109) of the patients without endometrial cancer had no or fewer than three genes methylated in their vaginal secretion. The methods developed in this study provide the basis for a prospective clinical trial to screen asymptomatic women who are at high risk for endometrial cancer.

Repeat surgery in patients with cervical cancer stage FIGO IA1: a series of 156 cases and a review of the literature.

BACKGROUND: Many physicians advocate repeat surgery after cervical conization with a diagnosis of cervical cancer stage FIGO IA1. In a multicenter trial, whether repeat surgery is a necessary therapeutic procedure in the treatment of cervical cancer stage FIGO IA1 was evaluated and a literature review performed. PATIENTS AND METHODS: From 1997 to 2006, 156 patients with squamous cell cervical cancer, stage FIGO IA1, were primarily treated with conization in three different institutions; 102 of these patients underwent repeat surgery, comprising the study group for the present trial. RESULTS: In the conization specimen, 22 patients had clear resection margins, none of whom had residual dysplasia in the repeat conization/hysterectomy specimen. Sixty-four patients had cervical intraepithelial neoplasia (CIN) I-III at the conization resection margin; of these, 29, 9, 24, and 2 patients had no sign of residual dysplasia, CIN I, CIN II/III, or multifocal cervical cancer FIGO IA1 in the repeat conization/hysterectomy specimen, respectively. Sixteen patients had invasive cancer at the resection margin of the conization specimen; no sign of dysplasia, CIN I, CIN II/III, or residual cervical cancer were found in the repeat conization/hysterectomy specimen in 4, 1, 5, and 6 cases, respectively. In a multivariate analysis, risk factors for residual CIN II/III or multifocal invasive carcinoma in patients with CIN at the resection margin were advanced patient age and presence of multifocal invasive cervical cancer, but not depth of invasion, lymphovascular space involvement (LVSI), nor positive endocervical curettage. CONCLUSION: The risk of residual dysplasia after conization of FIGO IA1 cervical cancer with clear margins is minimal. A considerable number of patients with locally resected FIGO IA1 cervical cancer, who had CIN I-III at the resection margin, showed signs of residual high-grade CIN or multifocal cervical cancer. The need for repeat surgery when signs of invasive carcinoma are present at the resection margins after conization is clear.

Ultrasound-guided core needle biopsy of the breast: does frozen section give an accurate diagnosis?

Reducing the period of uncertainty between the discovery of a breast tumor and histological diagnosis alleviates the psychological impact of breast cancer to an important degree. We aimed to verify whether histological results obtained with frozen sections of core needle biopsies (CNBs) offer an accurate and reliable tool for minimising this period. In 2619 cases we compared histological diagnosis on frozen sections with those on paraffin sections of CNB and finally with the results of open biopsies. Of the cases 49% were proved malignant and 51% benign. In 99.3% of the malignant lesions preceding CNB was correctly classified as B5 (n = 1185, 92.9%) or at least B4 (n = 82, 6.4%) in frozen and in paraffin sections. There were seven false-negative cases in frozen (false-negative rate = 0.5%) and five false-negative cases (false-negative rate = 0.4%) in paraffin sections of CNB. On frozen sections complete sensitivity was 99.5% and the positive predictive value of B5 was 99.9%. There was one false-positive case in frozen sections and one in paraffin sections. False-positive rate = 0.08%, negative predictive value for B2 = 99.4% for frozen and 99.6% for paraffin sections; full specificity was 85.9 for frozen and 85.8 for paraffin sections of CNBs. Immediate investigation of CNB in frozen sections is an accurate diagnostic method and an important step in reducing psychological strain on patients with breast tumors and may be offered by specialised Breast Assessment Units.