miR-181a and miR-630 Regulate Cisplatin-Induced Cancer Cell Death

• Published in: Cancer research (Chicago, Ill.) Vol. 70; no. 5; pp. 1793 - 1803
• Main Authors: GALLUZZI, Lorenzo, MORSELLI, Eugenia, RIPOCHE, Hugues, LAZAR, Vladimir, HAREL-BELLAN, Annick, DESSEN, Philippe, BARILLOT, Emmanuel, KROEMER, Guido, VITALE, Ilio, KEPP, Oliver, SENOVILLA, Laura, CRIOLLO, Alfredo, SERVANT, Nicolas, PACCARD, Caroline, HUPE, Philippe, ROBERT, Thomas
• Format: Journal Article
• Language: English
• Published: Philadelphia, PA American Association for Cancer Research 01.03.2010
• Subjects: Antineoplastic agents; Antineoplastic Agents - pharmacology; Apoptosis - drug effects; Apoptosis - genetics; bcl-2-Associated X Protein - metabolism; Biological and medical sciences; Cadmium Chloride - pharmacology; Carcinoma, Non-Small-Cell Lung - drug therapy; Carcinoma, Non-Small-Cell Lung - genetics; Carcinoma, Non-Small-Cell Lung - metabolism; Carcinoma, Non-Small-Cell Lung - pathology; Caspases - metabolism; Cell Cycle - drug effects; Cell Cycle - genetics; Cell Line, Tumor; Cisplatin - pharmacology; DNA Damage; HeLa Cells; Humans; Lung Neoplasms - drug therapy; Lung Neoplasms - genetics; Lung Neoplasms - metabolism; Lung Neoplasms - pathology; Medical sciences; MicroRNAs - genetics; MicroRNAs - metabolism; MicroRNAs - physiology; Oligonucleotide Array Sequence Analysis - methods; Pharmacology. Drug treatments; Phosphorylation; Sphingosine - analogs & derivatives; Sphingosine - pharmacology; Tumor Suppressor Protein p53 - genetics; Tumor Suppressor Protein p53 - metabolism; Tumors; Up-Regulation - drug effects; Antineoplastic agent; Gene silencing; Alkylating agent; RNA interference; Cell death; Micro RNA; Tumor cell; Cisplatin; Platinum II Complexes; Apoptosis
• ISSN: 0008-5472; 1538-7445
• DOI: 10.1158/0008-5472.can-09-3112

MicroRNAs (miRNA) are noncoding RNAs that regulate multiple cellular processes, including proliferation and apoptosis. We used microarray technology to identify miRNAs that were upregulated by non-small cell lung cancer (NSCLC) A549 cells in response to cisplatin (CDDP). The corresponding synthetic miRNA precursors (pre-miRNAs) per se were not lethal when transfected into A549 cells yet affected cell death induction by CDDP, C2-ceramide, cadmium, etoposide, and mitoxantrone in an inducer-specific fashion. Whereas synthetic miRNA inhibitors (anti-miRNAs) targeting miR-181a and miR-630 failed to modulate the response of A549 to CDDP, pre-miR-181a and pre-miR-630 enhanced and reduced CDDP-triggered cell death, respectively. Pre-miR-181a and pre-miR-630 consistently modulated mitochondrial/postmitochondrial steps of the intrinsic pathway of apoptosis, including Bax oligomerization, mitochondrial transmembrane potential dissipation, and the proteolytic maturation of caspase-9 and caspase-3. In addition, pre-miR-630 blocked early manifestations of the DNA damage response, including the phosphorylation of the ataxia-telangiectasia mutated (ATM) kinase and of two ATM substrates, histone H2AX and p53. Pharmacologic and genetic inhibition of p53 corroborated the hypothesis that pre-miR-630 (but not pre-miR-181a) blocks the upstream signaling pathways that are ignited by DNA damage and converge on p53 activation. Pre-miR-630 arrested A549 cells in the G0-G1 phase of the cell cycle, correlating with increased levels of the cell cycle inhibitor p27(Kip1) as well as with reduced proliferation rates and resulting in greatly diminished sensitivity of A549 cells to the late S-G2-M cell cycle arrest mediated by CDDP. Altogether, these results identify miR-181a and miR-630 as novel modulators of the CDDP response in NSCLC.