Molecular alterations in endometrial archived liquid‐based cytology

Abstract

Endometrial cancer is one of the most common gynecological malignancy worldwide and its prevalence is increasing. The introduction of liquid-based cytology (LBC) and endoflower dispositive in routine practice gives the possibility to examine endometrial cells by cytological diagnosis and may also release the opportunity to study molecular alterations, in endometrioid type cancer in which carcinogenesis is well known. We gathered 72 cases of endometrial LBC samples and corresponding formalin-fixed paraffin-embedded (FFPE) blocks, collected from 2004 to 2010. DNA was isolated from both samples using standard protocols. DNA quality and quantity were assessed using Nanodrop and BIOMED2 multiplex PCR. Mutations in exon 5 of PTEN and exon 20 of PI3K were studied using Sanger sequencing. DNA with good quality and amount was isolated from 67/72 FFPE cases. In these samples, two cases were found to harbor mutations in exon 5 of PTEN. No PI3K mutations were identified. LBC samples were then assessed to verify the concordance with the FFPE DNA results. The results obtained were concordant, that is the wild type cases in FFPE were also wild type in LBC and vice versa for the mutated case. Unfortunately, the second case of mutation in PTEN could not be confirmed in LBC due to low amount of DNA obtained. Detection of molecular alterations in LBC will open a new era for the detection in asymptomatic women of precursor lesions that could evolve into cancer and for endometrial cancer diagnosis and screening in selected high-risk women.

Expression of PTEN in Endometrial Liquid-Based Cytology

“…Recently, we demonstrated that it is possible to use archived endometrial LBC samples to detect molecular alterations with a good amount and high quality of DNA extracted. We showed the same PTEN alteration both in histology and in cytology [19]. However, the high costs and the limitations of applying molecular techniques in routine practice stimulated us to study the consistency of PTEN immunostaining in LBC samples.…”

“…In the clinical perspective, the main and important advantages of using this technique are the low cost, the easy performance in office-based surroundings and the minimal discomfort for patients. Many studies have demonstrated its high accuracy in detecting endometrial cancer, especially with the LBC method [16,17,18,19,20,21]. In fact, the high detection rate is due to the possibility of taking samples from the entire uterus cavity, improving the diagnostic rate of biopsy [18].…”

“…According to the literature and based on our own experience, the LBC residual material, after morphological evaluation, is also suitable for ancillary techniques [19,20,21]. Recently, we demonstrated that it is possible to use archived endometrial LBC samples to detect molecular alterations with a good amount and high quality of DNA extracted.…”

The Yokohama system for reporting directly sampled endometrial cytology: The quest to develop a standardized terminology

“…Tumors with no or rare and sparse positive cells, but in any case <50% of the cancer cells, were considered negative (Figures and ). All the normal endometrium control cases were positive in cytology and histology (Figure ) …”

Endometrial carcinoma in high‐risk populations: is it time to consider a screening policy?

“…The final score was diagnostically useful and the authors proposed that it could be used in pre‐operative cytological samples. Di Lorito et al . have demonstrated recently that it is possible to apply molecular techniques on endometrial LBC, and Norimatsu et al .…”