CNOT3 targets negative cell cycle regulators in non-small cell lung cancer development


Lung cancer is one of the major causes of cancer death and clarification of its molecular pathology is highly prioritized. The physiological importance of mRNA degradation through the CCR4-NOT deadenylase has recently been highlighted. For example, mutation in CNOT3, a gene coding for CNOT3 subunit of the CCR4-NOT complex, is found to be associated with T-cell acute lymphoblastic leukemia, T-ALL, though its contribution to other cancers has not been reported. Here, we provide evidence suggesting that CNOT3 is required for the growth of non-small cell lung cancer. Depletion of CNOT3 suppresses proliferation of A549 human non-small cell lung cancer cells with enhanced mRNA stability and subsequent elevated expression of p21. In addition, we identified the mRNA for Krüppel-like factor 2 transcription factor, an inducer of p21, as a novel mRNA degradation target of CNOT3 in non-small cell lung cancer cells. Aberrant up-regulation of Krüppel-like factor 2 by CNOT3 depletion leads to impairment in the proliferation of A549 cells. Consistent with these findings, elevated mRNA expression of CNOT3 in non-small cell lung cancer in comparison with the paired normal lung epithelium was confirmed through scrutinization of the RNA-sequencing datasets from The Cancer Genome Atlas. Moreover, we found an inverse correlation between CNOT3 and CDKN1A (encoding p21) mRNA expression using the combined datasets of normal lung epithelium and non-small cell lung cancer. Thus, we propose that the up-regulation of CNOT3 facilitates the development of non-small cell lung cancer through down-regulation of Krüppel-like factor 2 and p21, contrary to tumor suppressive functions of CNOT3 in T-ALL.

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The authors would like to thank Dr. Hiroyuki Miyoshi (RIKEN) for lentivirus vector system. The authors also thank all the members of Cell Signal Unit at the Okinawa Institute of Science and Technology Graduate University (OIST) for discussion and advice. This research was supported by subsidiary of OIST and KAKENHI Grants-in-Aid for scientific research (S) (21229006 to TY), for Scientific Research on Innovative Areas (25121734 to TY), for Young Scientists (B) (26830066 to AM and 16K18437 to MH), and for Research Activity Start-up (24890289 to Y-TS and 26893050 to MH) from the Japan Society for the Promotion of Science (JSPS).

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Correspondence to Yo-Taro Shirai or Tadashi Yamamoto.

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Shirai, Y., Mizutani, A., Nishijima, S. et al. CNOT3 targets negative cell cycle regulators in non-small cell lung cancer development. Oncogene 38, 2580–2594 (2019).

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