Multiple transcriptional events take place when normal urothelium is transformed into tumor tissue. These can now be monitored simultaneously by the use of oligonucleotide arrays, and expression patterns of superficial and invasive tumors can be established. Single-cell suspensions were prepared from bladder biopsies (36 normal, 29 tumor). Pools of cells were made from normal urothelium and from pTa grade I and II and pT2 grade III and IV bladder tumors. From these suspensions, and from 10 single-tumor biopsies, labeled cRNA was hybridized to oligonucleotide arrays carrying probes for 6500 genes. The obtained expression data were sorted according to a weighting scheme and were subjected to hierarchical cluster analysis of tissues and genes. Northern blotting was used to verify the array data, and immunohistology was used to correlate between RNA and protein levels. Hierarchical clustering of samples correctly identified the stage using both 4076 genes and a subset of 400 genes covarying with the stages and grades of tumors. Hierarchical clustering of gene expression levels identified several stage-characteristic, functionally related clusters, encoding proteins that were related to cell proliferation, oncogenes and growth factors, cell adhesion, immunology, transcription, proteinases, and ribosomes. Northern blotting correlated well with array data. Immunohistology showed a good concordance between transcript level and protein staining. The study indicates that gene expression patterns may be identified in bladder cancer by combining oligonucleotide arrays and cluster analysis. These patterns give new biological insight and may form a basis for the construction of molecular classifiers and for developing new therapy for bladder cancer.
"From transcriptomic data obtained on human BC biopsies, we found that RNA levels of TTK were highly increased in TNBC samples compared to the other BC subgroups and healthy tissue samples. Our results are in agreement with previous microarray analyses showing that TTK is up-regulated in a variety of tumors, such as breast, bladder, esophagus, lung, prostate and anaplastic thyroid , , –. Moreover, TTK belongs to a signature defined by the top 25 genes overexpressed in chromosomally instable (CIN) and aneuploid tumors , highlighting their “addiction” to the mitotic checkpoint to sustain cell proliferation in the presence of CIN and aneuploidy. "
[Show abstract][Hide abstract] ABSTRACT: Triple-negative breast cancer (TNBC) represents a subgroup of breast cancers (BC) associated with the most aggressive clinical behavior. No targeted therapy is currently available for the treatment of patients with TNBC. In order to discover potential therapeutic targets, we searched for protein kinases that are overexpressed in human TNBC biopsies and whose silencing in TNBC cell lines causes cell death. A cohort including human BC biopsies obtained at Institut Curie as well as normal tissues has been analyzed at a gene-expression level. The data revealed that the human protein kinase monopolar spindle 1 (hMPS1), also known as TTK and involved in mitotic checkpoint, is specifically overexpressed in TNBC, compared to the other BC subgroups and healthy tissues. We confirmed by immunohistochemistry and reverse phase protein array that TNBC expressed higher levels of TTK protein compared to the other BC subgroups. We then determined the biological effects of TTK depletion by RNA interference, through analyses of tumorigenic capacity and cell viability in different human TNBC cell lines. We found that RNAi-mediated depletion of TTK in various TNBC cell lines severely compromised their viability and their ability to form colonies in an anchorage-independent manner. Moreover, we observed that TTK silencing led to an increase in H2AX phosphorylation, activation of caspases 3/7, sub-G1 cell population accumulation and high annexin V staining, as well as to a decrease in G1 phase cell population and an increased aneuploidy. Altogether, these data indicate that TTK depletion in TNBC cells induces apoptosis. These results point out TTK as a protein kinase overexpressed in TNBC that may represent an attractive therapeutic target specifically for this poor prognosis associated subgroup of breast cancer.
PLoS ONE 05/2013; 8(5):e63712. DOI:10.1371/journal.pone.0063712 · 3.23 Impact Factor
"Several attempts and many criteria have been suggested for the diagnosis of dysplastic grades or cytologic changes from the basal cell layer and upward.[10–13] Expression of thousands of genes during tumor progression by array technologies which may form a basis for a detailed molecular characterization of cancer development. A recent impressive series of studies has focused on DNA ploidy measurements in patients with oral epithelial dysplasia during a rather long follow-up period.[16–18] "
[Show abstract][Hide abstract] ABSTRACT: E-cadherin is known to be an invasion suppressor gene and cathepsin-D, a protease, which is an invasion promoter and plays a central role in solid tumors including oral cancer.
To look for the expression pattern in normal buccal mucosa, dysplastic oral epithelium and oral squamous cell carcinoma (SCC) along with their correlation to individual atypical features, thereby providing an objective to the grading system in predicting the fate of affected epithelium.
To elucidate the expression patterns of these markers, we examined immunohistochemically on formalin fixed, paraffin embedded sections 22 dysplastic epithelia, eight SCC and ten normal buccal mucosa.
In dysplastic epithelium slight loss of expression of E-cadherin was noted as grade of dysplasia increased. Two cases of carcinoma clearance showed only basal and suprabasal staining. The staining varied in SCC with patchy to complete absence of expression. With cathepsin-D fine to moderate granular cytoplasmic staining was noted in most of the dysplastic epithelium. Similar staining was noted in SCC. The atypical features which strongly correlated to loss of expression of E-cadherin and intense cathepsin-D expression are basilar hyperplasia, loss of cohesion, mitosis, loss of polarity and drop shaped rete ridges.
The result of the study shows that the above atypical features should be given more weightage in addition to traditional grading system, in predicting the fate of affected epithelium. Additional studies with larger sample size and using monoclonal antibody against cathepsin-D may further strengthen our findings.
Journal of Oral and Maxillofacial Pathology 09/2011; 15(3):288-94. DOI:10.4103/0973-029X.86689
"Among our identified candidate migration/progression genes, versican, ephrin-B2 and E-cadherin are well known to be associated with tumor transformation and progression . We also identified E-cadherin and fibronectin 1 as biomarkers for both cell migration and tumor stage consistent with data from other microarrays (Thykjaer et al., 2001; Modlich et al., 2004). Elevated levels of ECM versican are also predictive of poor prognosis in patients with prostate cancer, endometrial cancer and oral squamous cell carcinoma (Ricciardelli et al., 1999; Kodama et al., 2007; Pukkila et al., 2007). "
[Show abstract][Hide abstract] ABSTRACT: Cell migration is essential to cancer invasion and metastasis and is spatially and temporally integrated through transcriptionally dependent and independent mechanisms. As cell migration is studied in vitro, it is important to identify genes that both drive cell migration and are biologically relevant in promoting invasion and metastasis in patients with cancer. Here, gene expression profiling and a high-throughput cell migration system answers this question in human bladder cancer. In vitro migration rates of 40 microarray-profiled human bladder cancer cell lines were measured by radial migration assay. Genes whose expression was either directly or inversely associated with cell migration rate were identified and subsequently evaluated for their association with cancer stage in 61 patients. This analysis identified genes known to be associated with cell invasion such as versican, and novel ones, including metallothionein 1E (MT1E) and nicotinamide N-methyltransferase (NNMT), whose expression correlated positively with cancer cell migration and tumor stage. Using loss of function analysis, we show that MT1E and NNMT are necessary for cancer cell migration. These studies provide a general approach to identify the clinically relevant genes in cancer cell migration and mechanistically implicate two novel genes in this process in human bladder cancer.
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