Brain metastasis development and poor survival associated with carcinoembryonic antigen (CEA) level in advanced non-small cell lung cancer: a prospective analysis

<p>Abstract</p> <p>Background</p> <p>Central nervous system is a common site of metastasis in NSCLC and confers worse prognosis and quality of life. The aim of this prospective study was to evaluate the prognostic significance of clinical-pathological factors (CPF), serum CEA levels, and EGFR and HER2 tissue-expression in brain metastasis (BM) and overall survival (OS) in patients with advanced NSCLC.</p> <p>Methods</p> <p>In a prospective manner, we studied 293 patients with NSCLC in IIIB-IV clinical stage. They received standard chemotherapy. CEA was measured prior to treatment; EGFR and HER2 were evaluated by immunohistochemistry. BM development was confirmed by MRI in symptomatic patients.</p> <p>Results</p> <p>BM developed in 27, and 32% of patients at 1 and 2 years of diagnosis with adenocarcinoma (RR 5.2; 95% CI, 1.002–29; p = 0.05) and CEA ≥ 40 ng/mL (RR 11.4; 95% CI, 1.7–74; <it>p </it>< 0.01) as independent associated factors. EGFR and HER2 were not statistically significant. Masculine gender (RR 1.4; 95% CI, 1.002–1.9; <it>p </it>= 0.048), poor performance status (RR 1.8; 95% CI, 1.5–2.3; <it>p </it>= 0.002), advanced clinical stage (RR 1.44; 95% CI, 1.02–2; <it>p </it>= 0.04), CEA ≥ 40 ng/mL (RR 1.5; 95% CI, 1.09–2.2; <it>p </it>= 0.014) and EGFR expression (RR 1.6; 95% CI, 1.4–1.9; <it>p </it>= 0.012) were independent associated factors to worse OS.</p> <p>Conclusion</p> <p>High CEA serum level is a risk factor for BM development and is associated with poor prognosis in patients with advanced NSCLC. Surface expression of CEA in tumor cells could be the physiopathological mechanism for invasion to CNS.</p

P4-04-02: Smoothened Function as a G-Protein Coupled Receptor in Mammary Epithelial Cells.

Abstract Background Hedgehog signaling orchestrates many key developmental processes, including cell fate determination and tissue patterning in the embryo, as well as tissue homeostasis and stem cell maintenance in the adult. Aberrant hedgehog signaling has been implicated in several cancers including skin, brain, prostate and breast. Previous work in our laboratory demonstrated that overexpression of activated Smoothened (SMO), a key effector in hedgehog signaling, leads to mammary hyperplasia, and is overexpressed in DCIS and IBCs. Consistent with recent data, we now provide evidence that SMO signals via Gαi proteins in the mouse mammary gland. Hypothesis: Smoothened promotes mammary gland proliferation by functioning as a G-protein coupled receptor(GPCR) and couples to one or more members of the PTX-sensitive Gαi family of heterotrimeric G-protein subunits. Experimental Design and Methods: Expression of Gαi subunit(s) was tested by qRT-PCR &amp; immunofluorescence microscopy of mammary epithelial cells (MECs) and whole mammary glands from SMO and WT mice. We analyzed mammary glands from single Gαi subunit null mice for developmental abnormalities. Genetic studies are underway in which SMO mice are crossed to Gαil2&amp;3 null mice. To elucidate downstream signaling events triggered by SMO activation, we are treating SMO derived MECs cultured in a 3D matrix and whole gland organ cultures with inhibitors of Gαi effector molecules and assessing proliferation status. We will also employ GTPγS binding assays using primary MECs from SMO overexpressing mice to test whether SMO can activate Gαi proteins in mammary gland epithelium. Results and Discussion: Treatment of MMTV-Cre dependent SMO hyperplasias with pertussis toxin (PTX), a potent inhibitor of Gαi family of G-protein signaling, significantly attenuates mammary gland hyperproliferation. This result supports the hypothesis that Smoothened functions as a GPCR interacting with one or more members of the Gαi family. We have now found that Gαi2 and Gαi3 null mice display an increased number of terminal end buds and a more completely filled fat pad relative to WT control littermates. Our data suggest that SMO-expressing cells induce proliferation of neighboring WT cells and that low-level proliferation in SMO-expressing cells is not affected by PTX. qPCR data indicates that Gαi1 is the most abundant transcript in sorted MECs which overexpress SMO and that the Notch signaling pathway may be activated. While our mechanistic data thus far mostly comprises transcriptional analyses coupled with pharmacological inhibition, they offer insight as to which pathways may be regulated by SMO activation. Genetic crosses between the SMO and Gαi null mice should provide evidence that these two proteins interact in the developing mammary gland. When associated with biochemical data, these studies ought to provide functional insight into the mechanism behind SMO-driven mammary hyperplasias. This work offers potential clinical implications towards breast cancer treatment as there exist therapeutic agents targeting the hedgehog signaling pathway in clinical trials that were developed by solely testing their ability to inhibit GLI1/2-mediated transcription. Our data identify targets for intervention other than the GLI transcription factors. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-04-02.</jats:p

An analysis of Cyclin D1, Cytokeratin 5/6 and Cytokeratin 8/18 expression in breast papillomas and papillary carcinomas

<p>Abstract</p> <p>Background</p> <p>To evaluate the expression levels of Cyclin D1 in breast papillomas and papillary carcinomas, and to analyze the types of cells that co-express Cyclin D1 with Cytokeratin 5/6 (CK 5/6) or with Cytokeratin 8/18(CK 8/18).</p> <p>Methods</p> <p>Fifty-nine cases of papillary lesions including 36 papillomas and 23 intracystic papillary carcinomas were examined. Cyclin D1, CK 5/6 and CK 8/18 expression levels were evaluated by double immunostaining.</p> <p>Results</p> <p>Cyclin D1 is highly expressed in papillary carcinomas (27.54% ± 15.43%) compared with papillomas (8.81% ± 8.41%, p < 0.01). Cyclin D1 is predominantly expressed in Cytokeratin 8/18- expressing cells, rather than in Cytokeratin 5/6-expressing cells, regardless of the type of lesion. In Papillomas, Cyclin D1 exhibited a mean 11.42% (11.42% ± 10.17%) co-expression rate with Cytokeratin 8/18 compared with a mean 2.50% (2.50% ± 3.24%) co-expression rate with Cytokeratin 5/6 (p < 0.01). In papillary carcinomas, Cyclin D1 exhibited a mean 34.74% (34.74% ± 16.32%) co-expression rate with Cytokeratin 8/18 compared with a co-expression rate of 0.70% (0.70% ± 0.93%) with Cytokeratin 5/6 (p < 0.01).</p> <p>Conclusions</p> <p>The increase in Cyclin D1 suggests an association of Cyclin D1 staining with papillary carcinomas. Although Cyclin D1 is an effective marker for the differential diagnosis of other papillary lesions, it cannot be used to distinguish between papilloma and papillary carcinoma lesions because its expression occurs in both lesions. Our results show that Cyclin D1 and CK 5/6 staining could be used in concert to distinguish between the diagnosis of papilloma (Cyclin D1 < 4.20%, CK 5/6 positive) or papillary carcinoma (Cyclin D1 > 37.00%, CK 5/6 negative). In addition, our data suggest that Cyclin D1 is expressed only in the cancer stem or progenitor cells that co-immunostained with CK 8/18 in papillary carcinomas, and predominantly with CK 8/18 in the papillomas.</p> <p>Virtual slides</p> <p>The virtual slide(s) for this article can be found here: <url>http://www.diagnosticpathology.diagnomx.eu/vs/7299340558756848</url></p

Non-cell autonomous control of apoptosis by ligand-independent Hedgehog signaling in Drosophila

Hedgehog (Hh) signaling is important for development and homeostasis in vertebrates and invertebrates. Ligand-independent, deregulated Hh signaling caused by loss of negative regulators such as Patched causes excessive cell proliferation, leading to overgrowth in Drosophila and tumors in humans, including basal-cell carcinoma and medulloblastoma. We show that in Drosophila deregulated Hh signaling also promotes cell survival by increasing the resistance to apoptosis. Surprisingly, cells with deregulated Hh activity do not protect themselves from apoptosis; instead, they promote cell survival of neighboring wild-type cells. This non-cell autonomous effect is mediated by Hh-induced Notch signaling, which elevates the protein levels of Drosophila inhibitor of apoptosis protein-1 (Diap-1), conferring resistance to apoptosis. In summary, we demonstrate that deregulated Hh signaling not only promotes proliferation but also cell survival of neighboring cells. This non-cell autonomous control of apoptosis highlights an underappreciated function of deregulated Hh signaling, which may help to generate a supportive micro-environment for tumor development