Project description:The Hippo pathway plays a key role in development, organ size control, and tissue homeostasis, and its dysregulation contributes to cancer. The LATS tumor suppressor kinases phosphorylate and inhibit the YAP/TAZ transcriptional co-activators to suppress gene expression and cell growth. Through a screen of marine natural products, we identified microcolin B (MCB) as a Hippo activator. Structure-activity optimization yielded more potent MCB analogs, which led to identification of PITP as the direct molecular targets. We established a critical role of PITP in regulating LATS and YAP. Moreover, we showed that PITP influences the Hippo pathway via cellular PI4P. This study uncovers a previously unrecognized role of PITP in Hippo pathway regulation and as potential cancer therapeutic targets.
Project description:Using this approach, we obtained phospho-proteomic data on the LATS1 interactome from cells treated with two proapoptotic signals: FAS and etoposide, which both activate LATS kinase activity [28].To identify the LATS1 interactome we transiently expressed GFP-LATS1 in HeLa cells, immunoprecipitated GFP-LATS1 with anti-GFP antibodies and identified the associated proteins using mass-spectrometry. Unspecific binding proteins were discarded by comparing with the control GFP IP.
Project description:To define the in vivo targets of the human nuclear exosome, we performed stranded RNA-seq using rRNA-depleted RNAs isolated from HeLa cells.
Project description:Analysis of STAT1 and IRF1 binding in IFNg treated and untreated HeLa cells for 6 hours was done using 50mer oligonucleotide probes at 30bp intervals tiling over non-repetitive 16MB gene locus (HG17).
Project description:Pharmacogenomic identification of targets for adjuvant therapy with the topoisomerase poison camptothecin. The response of tumor cells to the unusual form of DNA damage caused by topoisomerase poisons such as camptothecin (CPT) is poorly understood, and knowledge regarding which drugs can be effectively combined with CPT is lacking. To better understand the response of tumor cells to CPT and to identify potential targets for adjuvant therapy, we examined global changes in mRNA abundance in HeLa cells after CPT treatment using Affymetrix U133A GeneChips, which include all annotated human genes (22,283 probe sets). Statistical analysis of the data using a Bayesian/Cyber t test and a modified Benjamini and Hochberg correction for multiple hypotheses testing identified 188 probe sets that are induced and 495 that are repressed 8 h after CPT treatment at a False Discovery Rate of <0.05 and a minimum 3-fold change. This pharmacogenomic approach led us to identify two pathways that are CPT induced: (a) the epidermal growth factor receptor; and (b) nuclear factor-kappaB-regulated antiapoptotic factors. Experiments using HeLa cells in our lab and prior animal model studies performed elsewhere confirm that inhibitors of these respective pathways super-additively enhance CPT's cytotoxicity, suggesting their potential as targets for adjuvant therapy with CPT. Cancer Res. 2004 Mar 15;64(6):2096-104 Keywords = HeLa Keywords = Camptothecin Keywords: ordered
Project description:The objective of the present study is to investigate the role of processing glycosidase inhibition induced by processing glycosidase inhibitors, namely castanospermin as the processing glucosidases inhibitor, deoxymannojirimycin as the processing mannosidases inhibitor, and deoxyfuconojirimycin as the fucosidase inhibitor. Comparative gene expression analysis was performed in human cervical carcinoma HeLa cells, with non-inhibitor-treated cells as the negative control. Gene expression was analyzed using 3D-Gene® Human 25k oligonucleotide microarrays and computational gene expression analysis tools.
Project description:The objective of the present study is to investigate the role of processing glycosidase inhibition induced by processing glycosidase inhibitors, namely castanospermin as the processing glucosidases inhibitor, deoxymannojirimycin as the processing mannosidases inhibitor, and deoxyfuconojirimycin as the fucosidase inhibitor. Comparative gene expression analysis was performed in human cervical carcinoma HeLa cells, with non-inhibitor-treated cells as the negative control. Gene expression was analyzed using 3D-Gene® Human 25k oligonucleotide microarrays and computational gene expression analysis tools. Inhibition of processing glycosidases (glucosidases, mannosidases, and fucosidase) in human cervical carcinoma HeLa cells was performed by treating with a small molecule inhibitor and then culturing at 37°C. Total RNA samples were prepared from the cells. Gene expression was analyzed using 3D-Gene® Human 25k oligonucleotide microarrays. Sample preparation for array hybridization was carried out as described in the manufacturer's instructions.
