MicroRNA-200c and microRNA-31 regulate proliferation, colony formation, migration and invasion in serous ovarian cancer.

BACKGROUND: Serous epithelial ovarian cancer (SEOC) is a highly metastatic disease and its progression has been implicated with microRNAs. This study aimed to identify the differentially expressed microRNAs in Malaysian patients with SEOC and examine the microRNAs functional roles in SEOC cells. METHODS: Twenty-two SEOC and twenty-two normal samples were subjected to miRNA expression profiling using the locked nucleic acid (LNA) quantitative real-time PCR (qPCR). The localization of miR-200c was determined via LNA in situ hybridization (ISH). Functional analysis of miR-200c and miR-31 on cell proliferation, migration and invasion and clonogenic cell survival were assessed in vitro. The putative target genes of the two miRNAs were predicted by miRWalk program and expression of the target genes in SEOC cell lines was validated. RESULTS: The miRNA expression profiling revealed thirty-eight significantly dysregulated miRNAs in SEOC compared to normal ovarian tissues. Of these, eighteen were up-regulated whilst twenty miRNAs were down-regulated. We observed chromogenic miR-200c-ISH signal predominantly in the cytoplasmic compartment of both epithelial and inflammatory cancer cells. Re-expression of miR-200c significantly increased the cell proliferation and colony formation but reduced the migration and invasion of SEOC cells. In addition, miR-200c expression was inversely proportionate with the expression of deleted in liver cancer-1 (DLC-1) gene. Over-expression of miR-31 in SEOC cells resulted in decreased cell proliferation, clonogenic potential, cell migration and invasion. Meanwhile, miR-31 gain-of-function led to the down-regulation of AF4/FMR2 family member 1 (AFF1) gene. CONCLUSIONS: These data suggested that miR-200c and miR-31 may play roles in the SEOC metastasis biology and could be considered as promising targets for therapeutic purposes.

Adaptive localization of focus point regions via random patch probabilistic density from whole-slide, Ki-67-stained brain tumor tissue.

Analysis of whole-slide tissue for digital pathology images has been clinically approved to provide a second opinion to pathologists. Localization of focus points from Ki-67-stained histopathology whole-slide tissue microscopic images is considered the first step in the process of proliferation rate estimation. Pathologists use eye pooling or eagle-view techniques to localize the highly stained cell-concentrated regions from the whole slide under microscope, which is called focus-point regions. This procedure leads to a high variety of interpersonal observations and time consuming, tedious work and causes inaccurate findings. The localization of focus-point regions can be addressed as a clustering problem. This paper aims to automate the localization of focus-point regions from whole-slide images using the random patch probabilistic density method. Unlike other clustering methods, random patch probabilistic density method can adaptively localize focus-point regions without predetermining the number of clusters. The proposed method was compared with the k-means and fuzzy c-means clustering methods. Our proposed method achieves a good performance, when the results were evaluated by three expert pathologists. The proposed method achieves an average false-positive rate of 0.84% for the focus-point region localization error. Moreover, regarding RPPD used to localize tissue from whole-slide images, 228 whole-slide images have been tested; 97.3% localization accuracy was achieved.

Estrogen receptor-negative breast ductal carcinoma: clinicopathological features and MIB-1 (Ki-67) proliferative index association.

Breast cancer estrogen receptor (ER) status is one of the strong additional factors in predicting response of patients towards hormonal treatment. The main aim of this study was to assess the morphological characteristics and proliferative activity using MIB-1(Ki-67) of estrogen receptor negative invasive breast ductal carcinoma (NOS type) as well as to correlate these features with clinicopathological data. We also aim to study the expression of c-erbB2 in ER negative breast tumors. High proliferative rate (MIB-1 above 20%) was observed in 63 (63.6%) of 99 ER negative tumors and that these tumors were associated with high expression of c-erbB2 (57.6%). We observed that MIB-1 is a reliable independent prognostic indicator for ER negative infiltrating ductal carcinoma in this study.

Perilobar nephrogenic rests and chromosome 22.

Perilobar nephrogenic rests (NR) are precursor lesions that may display genetic changes similar to their associated Wilms tumor (WT). Two patients presented with WT, both with perilobar NR and 1 with bilateral, multifocal metachronous WT. Both patients' WT displayed monosomy 22 as the predominant cytogenetic change, and the constitutional cytogenetic analysis was normal. The purpose of our study was to identify at what stage in the morphologic progression from NR to WT the monosomy 22 occurred by using a fluorescent in situ hybridization probe for chromosome 22 in the subtypes of perilobar NR and WT present in both cases. Section and core fluorescent in situ hybridization with a chromosome 22 probe was performed on formalin-fixed, paraffin-embedded tissues containing WT and perilobar NR. We also performed fluorescent-based microsatellite analysis on some of the WT in the bilateral case to determine whether there was a preferential loss of the same allele of chromosome 22. Sclerotic and dormant perilobar NR showed a rate of monosomy 22 of only approximately 30%, but this increased to approximately 50% in hyperplastic and adenomatous NR. Monosomy 22 was present in 60%-80% of nuclei in WT. Microsatellite analysis showed loss of homozygosity, with preferential loss of the same allele of chromosome 22 in the tumors examined. There are differences in the rate of detection of monosomy 22 in perilobar NR and WT, suggesting loss of chromosome 22 in the progression of perilobar NR to WT in a subset of tumors.

Cytogenetic findings in Wilms' tumour: a single institute study.

AIMS: Cytogenetic abnormalities of Wilms' tumour (WT) treated in a single institution in Western Australia were reviewed. Correlation with histologic subtypes, stage, presence of nephrogenic rests and age of the patient at diagnosis were also evaluated. METHODS: 53 WT specimens were encountered between 1995 and 2009. Tissue culture was obtained in 49 (92%) specimens. Reports documenting histopathological features of the tumour, stage and outcome were also retrieved. RESULTS: A total of 53 tumour specimens from 42 patients/cases were examined and staged in accordance with the National Wilms' Tumor Study (NWTS). Thirty-eight cases were unilateral (34 unifocal, 4 multifocal) and four were bilateral (2 multifocal). Fifty tumours showed favourable histology WT. One tumour was a cystic partially differentiated nephroblastoma (CPDN). Two tumours showed diffuse anaplasia. Eighteen specimens had nephrogenic rests, seven with perilobar rests, 10 with intralobar rests and one with both types of nephrogenic rests. Twelve WTs were assigned as stage 1, 22 as stage 2, 16 as stage 3, and two each for stages 4 and 5. For chromosomal analysis, 92% of the specimens yielded results, of which 70% showed abnormal karyotype and 22% displayed normal karyotypic findings. Hyperdiploidy was more common than hypodiploidy. The most common chromosomal gain involved chromosome 12. Low stage tumours tended to have abnormal karyotypes with hyperdiploidy and hypodiploidy being more common. There was no statistical correlation between abnormal karyotype and stage or abnormal karyotype and age group or abnormal karyotype and non-blastemal WT. Eleven (58%) tumours harbouring nephrogenic rests displayed an abnormal karyotype. 16q loss or der(16)t(1;16) were more common in younger patients but no association was made between this chromosomal abnormality and stage. Monosomy 22 and gain of 1q were more common in older patients. Furthermore, monosomy 22 tended to occur in tumours of earlier stage. No correlation between 11p deletions and age or stage was seen. CONCLUSIONS: WTs are karyotypically heterogeneous tumours. Conventional cytogenetic analysis of WTs still remains a useful technique to assist in the understanding of these tumours.