CIP4 promotes lung adenocarcinoma metastasis and is associated with poor prognosis.

Aberrant epidermal growth factor receptor (EGFR) signaling in non-small cell lung cancer (NSCLC) is linked to tumor progression, metastasis and poor survival rates. Here we report the role of Cdc42-interacting protein 4 (CIP4) in the regulation of NSCLC cell invasiveness and tumor metastasis. CIP4 was highly expressed in a panel of NSCLC cell lines and normal lung epithelial cell lines. Stable knockdown (KD) of CIP4 in lung adenocarcinoma H1299 cells, expressing wild-type EGFR, led to increased EGFR levels on the cell surface and defects in sustained activation of Erk kinase in H1299 cells treated with EGF. CIP4 localized to leading edge projections in NSCLC cells, and CIP4 KD cells displayed defects in EGF-induced cell motility and invasion through extracellular matrix. This correlated with reduced expression and activity of matrix metalloproteinase-2 (MMP-2) in CIP4 KD cells compared with control. In xenograft assays, CIP4 silencing had no effect on tumor growth but resulted in significant defects in spontaneous metastases to the lungs from these subcutaneous tumors. This correlated with reduced expression of the Erk target gene Zeb1 and the Zeb1 target gene MMP-2 in CIP4 KD tumors compared with control. CIP4 also enhanced rates of metastasis to the liver and lungs in an intrasplenic experimental metastasis model. In human NSCLC tumor sections, CIP4 expression was elevated greater than or equal to twofold in 43% of adenocarcinomas and 32% of squamous carcinomas compared with adjacent normal lung tissues. Analysis of microarray data for NSCLC patients also revealed that high CIP4 transcript levels correlated with reduced overall survival. Together, these results identify CIP4 as a positive regulator of NSCLC metastasis and a potential poor prognostic biomarker in lung adenocarcinoma.

Transducer of Cdc42-dependent actin assembly promotes breast cancer invasion and metastasis.

Metastatic breast adenocarcinomas display activation signatures for signaling pathways that trigger cell motility and tissue invasion. Here, we report that the adaptor protein transducer of Cdc42-dependent actin assembly-1 (Toca-1) is expressed in highly invasive breast cancers and regulates their metastatic phenotypes. We show that Toca-1 localizes to the filamentous actin-rich core of invadopodial protrusions actively degrading the extracellular matrix (ECM). Toca-1 colocalizes with Cortactin, and we show that this interaction is mediated by the SH3 domain of Toca-1. Stable knockdown (KD) of Toca-1 expression in MDA-MB-231 cells led to a significant defect in epidermal growth factor (EGF)-induced cell migration and invasion. Toca-1 KD cells also showed significant defects in EGF- and Src-induced ECM digestion and formation of invadopodial membrane protrusions. To test the role of Toca-1 in metastasis, we achieved stable Toca-1 KD in both human and rat metastatic breast adenocarcinoma cell lines. Orthotopic tumor xenografting of control and Toca-1 KD cells in natural-killer /B-/T-cell-deficient mice revealed a significant defect in spontaneous lung metastases with Toca-1 silencing in vivo. In contrast, no defects in primary tumor growth or lung seeding following tail vein injection of Toca-1 KD cells was observed, suggesting that Toca-1 functions at an early step in the dissemination of metastatic breast tumor cells. Taken together, our results identify Toca-1 as a proinvasive protein in breast adenocarcinoma and a potential therapeutic target to limit tumor metastasis.

Molecular characterization of a cDNA from the true armyworm Pseudaletia unipuncta encoding Manduca sexta allatotropin peptide(1).

Allatotropin (AT) is an insect neuropeptide isolated from the tobacco hornworm, Manduca sexta, stimulates juvenile hormone (JH) biosynthesis by the corpora allata. A cDNA isolated from the true armyworm, Pseudaletia unipuncta, encodes a 135 amino acid AT precursor peptide which contains the AT peptide, with processing sites necessary for its endoproteolytic cleavage and amidation, plus two additional peptides of unknown function. The encoded AT peptide is identical to that isolated from M. sexta and Agrius convolvuli. Southern blot analysis indicated that AT is a single copy gene per haploid genome and is present in two allelic forms. A single transcript of approximately 1.5 kilobases was detected by northern blot analysis. The expression of the AT gene was analyzed during development from sixth instar larvae to five day-old moths. Initial expression was observed in late pupae and this expression was maintained throughout the adult stages in both sexes. In one day-old moths, expression was at its lowest level of the stages that express AT mRNA but levels increased in day 3 and day 5 adults. This pattern of AT expression in adult P. unipuncta moths mirrors that of JH biosynthesis and supports the notion that AT may act in the adult stages. Immunohistochemistry and in situ hybridization revealed that AT expression was localized to numerous structures of the nervous system, suggesting that AT may have functions distinct from regulation of JH biosynthesis.

Emotional support for patients with cancer who are undergoing CT: semistructured interviews of patients at a cancer institute.

To understand and improve the experience of cancer patients undergoing computed tomography (CT), 79 patients who underwent CT at a cancer institute participated in semistructured interviews about their experiences with CT. All patients had previously undergone CT; 75% (n = 59), three times or more. Anxiety about results was the most common concern during first and subsequent CT examinations. Technical aspects were a common concern during initial scanning, but not subsequently. Methods of relaxation most used by patients during CT were following instructions (56% [n = 44]), meditating and visualizing (44% [n = 35]), and praying (42% [n = 33]). Patients suggested several ways in which the radiology staff can support them during the evaluation of their malignancy. Fifty-five (70%) of the patients said they would like the radiologist to tell them the results of their scanning. Optimal care of patients with cancer who undergo CT goes beyond technical to emotional and spiritual support.