ABSTRACT
Cervical cancer with co-existing pathologic components of squamous cell carcinoma, basaloid morphology and sarcomatoid carcinoma is rare, with limited reports in the literature. Here we present a patient who underwent a modified radical hysterectomy for cervical cancer, with final pathology specimen demonstrating multiple histologic variants including basal carcinoma, adenoid cystic-like areas, basaloid squamous cell carcinoma and areas of high-grade transformation to sarcomatoid carcinoma.
ABSTRACT
TAZ (WWTR1) and YAP are transcriptional coactivators and oncoproteins inhibited by the Hippo pathway. Herein we evaluate 159 sarcomas representing the most prevalent sarcoma types by immunohistochemistry for expression and activation (nuclear localization) of TAZ and YAP. We show that 50% of sarcomas demonstrate activation of YAP while 66% of sarcomas demonstrate activated TAZ. Differential activation of TAZ and YAP are identified in various sarcoma types. At an RNA level, expression of WWTR1 or YAP1 predicts overall survival in undifferentiated pleomorphic sarcoma and dedifferentiated liposarcoma. Immunohistochemistry demonstrates that TAZ and YAP expression and activation are positively correlated with grade in the well-differentiated liposarcoma to dedifferentiated liposarcoma tumor progression sequence as well as conventional chondrosarcomas. TAZ and YAP are constitutively activated oncoproteins in sarcoma cell lines. Knock-down of TAZ and YAP demonstrate differential activity for the two proteins. Verteporfin decreases colony formation in soft agar as well as CTGF expression in sarcoma cell lines harboring activated TAZ and YAP.
Subject(s)
Adaptor Proteins, Signal Transducing/metabolism , Antineoplastic Agents/pharmacology , Carcinogenesis/metabolism , DNA-Binding Proteins/metabolism , Intracellular Signaling Peptides and Proteins/metabolism , Muscle Proteins/metabolism , Oncogene Proteins/metabolism , Phosphoproteins/metabolism , Porphyrins/pharmacology , Sarcoma/metabolism , Transcription Factors/metabolism , Adaptor Proteins, Signal Transducing/genetics , Antineoplastic Agents/therapeutic use , Cell Line, Tumor , Cell Nucleus/metabolism , Cell Proliferation/drug effects , Connective Tissue Growth Factor/metabolism , Disease Progression , Gene Knockdown Techniques , Humans , Immunohistochemistry , Intracellular Signaling Peptides and Proteins/genetics , Neoplasm Grading , Oncogene Proteins/genetics , Phosphoproteins/genetics , Protein Serine-Threonine Kinases , RNA Interference , RNA, Small Interfering , Sarcoma/drug therapy , Sarcoma/pathology , Signal Transduction/drug effects , TEA Domain Transcription Factors , Tissue Array Analysis , Trans-Activators , Transcriptional Coactivator with PDZ-Binding Motif Proteins , Verteporfin , YAP-Signaling ProteinsABSTRACT
We set out to find the "fenestrin" gene, a gene whose protein is associated with numerous cellular apertures, including the nuclear exchange junction in mating Tetrahymena thermophila. First we developed protocols for imaging and isolating intact nuclear exchange junctions from conjugating cells. Proteins from these junctions were purified using SDS-PAGE, subjected to limited proteolysis, and precise molecular weights were determined by mass spectrometry. Using Protein Prospector software and the published Tetrahymena Genome Database, genes for 15 of the most abundant proteins found in our extracts were identified. The most promising candidate was cloned by PCR, fused to yellow fluorescent protein (YFP), and placed under the control of an inducible metallothionein promoter. YFP-localization within live Tetrahymena transformants strongly suggested that one of these genes encoded the fenestrin protein, a result that was subsequently confirmed by Western blotting.
