Lung cancer is recognized as a leading cause of cancer-related deaths worldwide. Over the past several years, evidence emerged that microRNAs (miRNAs), a class of small non-coding RNA molecules regulating gene expression at posttranscriptional level, play an important role in cell functioning, as well as in human diseases. Here, we analyzed expression of miR-15a/16, miR-21, miR-34a, miR-126, miR-128, and miR-210 at transcriptional level in 30 non-small-cell lung carcinoma (NSCLC) tumor tissues compared to the matched adjacent normal tissues and their correlation with clinicopathological features of the patients. Samples were collected from the NSCLC patients undergoing surgery before radiotherapeutic or chemotherapeutic treatment. Expression levels of miRNAs were assessed by TaqMan RT-PCR assay. The data obtained in this study were processed using REST 2009 and SPSS statistical software. The graphs were designed by GraphPad prism 5.0. In tumor samples, we found downregulation of miR-15a/16 (50/83.3%), miR-34a (83.3%), miR-126 (70%), and miR-128 (63.3%). At the same time, miR-21 and miR-210 were upregulated by 53.3 and 66.6% in cancer tissue versus matched adjacent normal tissues, respectively. No significant correlation was found between the expression levels of miR-15a/16, miR-21, miR-34a, miR-126, miR-128, and miR-210 and lymph node, tumor size, sex, and smoking. However, the study demonstrated a correlation between a change in expression of miR-15, miR-16, miR-34a, miR-126, and miR-210 compared to normal tissues and TNM staging (P < 0.05). Furthermore, miR-126 expression level was different in adenocarcinomas and squamous cell carcinoma (SCC) subtype (P < 0.1). Detailed analysis revealed significant change in expression of miR-15a/16, miR-34a, miR-126, and miR-210 in NSCLC tumor samples indicating involvement of these miRNAs in lung cancer pathogenesis. miR-210 demonstrated the most consistent increase in tumor tissues between different patients, suggesting its potential significance for NSCLC.

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(2015) Upregulation of ATG3 contributes to autophagy induced by the detachment of intestinal epithelial cells from the extracellular matrix, but promotes autophagy-independent apoptosis of the attached cells, Autophagy, 11:8, 1230-1246, DOI: 10.1080/15548627.2015 Keywords: autophagy, apoptosis, anoikis, ATG3, ATG7, extracellular matrixAbbreviations: ATG, autophagy-related; BCL2L1, Bcl2-like1; CDK4, cyclin-dependent kinase 4; ECM, extracellular matrix; EGFR, epidermal growth factor receptor; GABARAP, GABA(A) receptor-associated protein; GFP, green fluorescent protein; LC3/ MAP1LC3, microtubule-associated protein 1 light chain 3; PE, phosphatidylethanolamine; qPCR, quantitative polymerase chain reaction; SD, standard deviation; SH3GLB1/Bif1, SH3-domain GRB2-like endophilin B1; siRNA, small interfering RNA; UVRAG, UV radiation resistance-associated.Detachment of nonmalignant intestinal epithelial cells from the extracellular matrix (ECM) triggers their growth arrest and, ultimately, apoptosis. In contrast, colorectal cancer cells can grow without attachment to the ECM. This ability is critical for their malignant potential. We found previously that detachment-induced growth arrest of nonmalignant intestinal epithelial cells is driven by their detachment-triggered autophagy, and that RAS, a major oncogene, promotes growth of detached cells by blocking such autophagy. In an effort to identify the mechanisms of detachment-induced autophagy and growth arrest of nonmalignant cells we found here that detachment of these cells causes upregulation of ATG3 and that ATG3 upregulation contributes to autophagy and growth arrest of detached cells. We also observed that when ATG3 expression is artificially increased in the attached cells, ATG3 promotes neither autophagy nor growth arrest but triggers their apoptosis. ATG3 upregulation likely promotes autophagy of the detached but not that of the attached cells because detachment-dependent autophagy requires other detachmentinduced events, such as the upregulation of ATG7. We further observed that those few adherent cells that do not die by apoptosis induced by ATG3 become resistant to apoptosis caused by cell detachment, a property that is critical for the ability of normal epithelial cells to become malignant. We conclude that cell-ECM adhesion can switch ATG3 functions: when upregulated in detached cells in the context of other autophagy-promoting events, ATG3 contributes to autophagy. However, when overexpressed in the adherent cells, in the circumstances not favoring autophagy, ATG3 triggers apoptosis.

Lung cancer is the leading cause of cancer-related deaths worldwide. Non-small cell lung cancer (NSCLC), the major lung cancer subtype, is characterized by high resistance to chemotherapy. Here we demonstrate that Tudor staphylococcal nuclease (SND1 or TSN) is overexpressed in NSCLC cell lines and tissues, and is important for maintaining NSCLC chemoresistance. Downregulation of TSN by RNAi in NSCLC cells led to strong potentiation of cell death in response to cisplatin. Silencing of TSN was accompanied by a significant decrease in S100A11 expression at both mRNA and protein level. Downregulation of S100A11 by RNAi resulted in enhanced sensitivity of NSCLC cells to cisplatin, oxaliplatin and 5-fluouracil. AACOCF3, a phospholipase A2 (PLA2) inhibitor, strongly abrogated chemosensitization upon silencing of S100A11 suggesting that PLA2 inhibition by S100A11 governs the chemoresistance of NSCLC. Moreover, silencing of S100A11 stimulated mitochondrial superoxide production, which was decreased by AACOCF3, as well as N-acetyl-L-cysteine, which also mimicked the effect of PLA2 inhibitor on NSCLC chemosensitization upon S100A11 silencing. Thus, we present the novel TSN-S100A11-PLA2 axis regulating superoxide-dependent apoptosis, triggered by platinum-based chemotherapeutic agents in NSCLC that may be targeted by innovative cancer therapies.

Resistance of carcinoma cells to anoikis, apoptosis that is normally induced by detachment of nonmalignant epithelial cells from the extracellular matrix, is thought to be critical for carcinoma progression. Molecular mechanisms that control anoikis of nonmalignant and cancer cells are understood poorly. In an effort to understand them we found that detachment of nonmalignant intestinal epithelial cells triggers upregulation of Chk2, a pro-apoptotic protein kinase that has never been implicated in anoikis and has been thought to kill cells mainly under the conditions compromising genome integrity. We found that enforced downregulation of Chk2 protects intestinal epithelial cells from anoikis. Chk2 can kill cells by stabilizing p53 tumor suppressor protein or via p53-independent mechanisms, and we established that Chk2-mediated anoikis of intestinal epithelial cells is p53-independent. We further found that, unlike nonmalignant intestinal epithelial cells whose anoikis is triggered by detachment-induced Chk2 upregulation, intestinal epithelial cells carrying oncogenic ras, a known inhibitor of anoikis, remain anoikis-resistant in response to enforced Chk2 upregulation. By contrast, drugs, such as topoisomerase I inhibitors, that can kill cells via Chk2-indpendent mechanisms, efficiently triggered anoikis of rastransformed cells. Thus, oncogenic ras can prevent Chk2 from triggering anoikis even when levels of this protein kinase are elevated in cancer cells, and the use of therapeutic agents that kill cells in a Chk-2-independent, rather than Chk-2-dependent, manner could represent an efficient strategy for overcoming ras-induced anoikis resistance of these cells. We conclude that Chk-2 is an important novel component of anoikis-promoting machinery of intestinal epithelial cells.