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Cytoskeleton proteins are substrates for proteases and further apoptotic death. We evaluated the participation of cytoskeleton in morphological changes during cell death induced by two apoptotic conditions, potassium deprivation (K5) and staurosporine, in cerebellar granule neurons (CGC). We found that K5 induced somatic damage, but neurites were relatively preserved, which corresponded to the reorganization of actin and α-tubulin in neurites. Staurosporine (STS) induced an early alteration of neurites with reorganization of cytoskeleton proteins in somas. Caspase inhibitor ZVAD totally inhibited STS-induced α-tubulin reorganization and partially blocked STS-induced actin reorganization. α-tubulin and actin reorganization induced by K5 was affected by ZVAD. Calpain inhibitor (IC1) did not affect α-tubulin or actin reorganization induced by STS, K5 or ionomycin. Neither ZVAD, nor IC1 changed α-tubulin or actin levels upon K5 treatment. STS increased α-tubulin and actin levels, but neither ZVAD nor IC1 changed α-tubulin levels upon STS treatment. In contrast, ZVAD reduced the STS-induced increase of actin. These results suggest that CGC cytoskeleton proteins undergo a differential expression and reorganization depending on the apoptotic condition.

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624 Background: Neoadjuvant chemoradiotherapy (nCRT) follow by surgery, as treatment for locally advanced rectal carcinoma (LARC), has improve local disease control, increase of complete pathological response (cPR) and preserve anal sphincter. cPR is associated with better prognosis. CRT act mainly through induction of DNA damage. MicroRNAs (miRNAs) are small non-coding RNAs able to regulate gene expression at post-transcriptional level. miRNAs are involved in the regulation of DNA damage/repair mechanism. Our goal was to measure expression of miRNAs involved in DNA damage/repair genes and correlated with cPR. Methods: Retrospectively, we analyzed the initial paraffin embedded tissue block of 20 Mexican patients with LARC treated with nCRT at National Cancer Institute of Mexico between 2010-2013. Treatment response was evaluated with Ryan Classification. Ten patients had cPR (Ryan Grade 0) meanwhile other 10 poor response (Ryan Grade 3). RNA extraction was done with RNeasy FFPE (Qiagen) Kit. RNA concentration and purity was assessed using NanoDrop 2000 Spectrophotometer. miRNAs expression were evaluated by real-time polymerase chain reaction (PCR) analysis with TaqMan Probes from Applied Biosystems. Statistical analysis was carried out with IBM SPSS Statistics 22.0. Differential expression between the two groups was performed with Chi square test. Results: There wasn’tsignificant difference betweenclinical/demographic features between the 2 groups. All patients received CRT at a total dose of 4500 cGy of pelvic irradiation, concomitantly with fluoropyrimidine. Surgical treatment was performed 14 weeks after completion of nCRT. The 2 miRNAs (188-5p and 590-5p) was overexpressed in the cPR group vs poor response group (p = 0.011 and p = 0.057, respectively). Conclusions: We evaluated the expression of miR590-5p and miR188-5p because they are related to DNA repair genes such as: MSH2, ERCC3, BRCC3 and XRCC5. We found overexpression in both miRNAs. Even though miR590 5p didn't reach statistical significance this maybe because of simple size.Our results now are been analyzed with a biological and functional genes network platform: Ingenuity Pathway Analysis.

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Lung cancer is the major cause of cancer-related deaths and has a poor prognosis with a 5-yr overall survival of <10%, mostly due to lack of early stage diagnosis. Cigarette smoking is estimated to be responsible for >80% of the cases. Previous studies have demonstrated that former smokers have a higher risk of lung cancer compared to nonsmokers. Early detection of lung cancer would improve the overall survival of this disease. However, there is no validated screening test for lung cancer. To develop an early detection test, we selected saliva as the biofluid of choice due to its accessibility, protein content that provides diagnostic information on a variety of diseases, including cancer. To identify differentially expressed proteins in saliva from individuals with lung adenocarcinoma, whole saliva samples were collected from 6 patients with lung adenocarcinoma and 6 matched healthy controls (male individuals, >40 years, smokers >10 pack year). Proteins and peptides from whole saliva samples were analyzed by two-dimensional gel electrophoresis identifying 20 differentially expressed protein spots. Mass spectrometry analysis identified 63 peptides and revealed fourteen non-redundant proteins in saliva obtained from lung adenocarcinoma patients. Four of these proteins have been previously reported as serum biomarkers of cancer, and two of them were associated with non-small cell lung cancer. These potential biomarkers are being tested in an independent and larger cohort for validation. Patient-based saliva proteomics is a promising approach to searching for cancer biomarkers. Further characterization of these markers may provide the basis for new, noninvasive tests for screening, detection, and monitoring of high-risk individuals. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4786. doi:1538-7445.AM2012-4786

e18028 Background: Brain metastases (BM) occur in 30-50% of non-small cell lung cancer (NSCLC) patients and confer worse prognosis and quality of life. Better selection of in-risk patients through an accurate biomarker could improve the benefit of prophylactic therapies. The aim of this prospective study was to determine a gene-expression profile (GEP) of primary tumor (PT) associated with BM in patients with advanced NSCLC. Methods: From January 2009 to June 2011, patients with stage IV of NSCLC were evaluated. PT core biopsy was performed prior to any treatment and snap-frozen. Samples with tumor cellularity > 70% and RNA integrity number > 8 were chosen for RNA isolation. A cDNA microarray platform representing 33,297 genes was used to obtain GEPs. All patients received standard chemotherapy. BM were confirmed by magnetic resonance imaging. Non- and supervised hierarchical clustering methods were employed to identify GEPs. Results: A total of 29 patients were enrolled, 79.4% (23/29) were adenocarcinomas, and 20.6% have other histology. BM were present in 15 (51.7%) patients, 14 at diagnosis and 1 was developed at 5 months of follow-up. Clinical characteristics were similar for patients with and without BM. At non- and supervised analyses, 35 genes up and down regulated were evidenced in BM group. From these, 11 transcripts with proteomic functions previously associated with metastasis processes were identified. Conclusions: Our work provides valuable biological information for development of predictive biomarkers for metastatic brain tumors from primary NSCLC. External validation of our gene-expression signature in a different set of patients is warranted. [Table: see text]