PMID- 37340282 OWN - NLM STAT- MEDLINE DCOM- 20230622 LR - 20230710 IS - 1420-9071 (Electronic) IS - 1420-682X (Print) IS - 1420-682X (Linking) VI - 80 IP - 7 DP - 2023 Jun 20 TI - DIS3L2 knockdown impairs key oncogenic properties of colorectal cancer cells via the mTOR signaling pathway. PG - 185 LID - 10.1007/s00018-023-04833-5 [doi] LID - 185 AB - DIS3L2 degrades different types of RNAs in an exosome-independent manner including mRNAs and several types of non-coding RNAs. DIS3L2-mediated degradation is preceded by the addition of nontemplated uridines at the 3'end of its targets by the terminal uridylyl transferases 4 and 7. Most of the literature that concerns DIS3L2 characterizes its involvement in several RNA degradation pathways, however, there is some evidence that its dysregulated activity may contribute to cancer development. In the present study, we characterize the role of DIS3L2 in human colorectal cancer (CRC). Using the public RNA datasets from The Cancer Genome Atlas (TCGA), we found higher DIS3L2 mRNA levels in CRC tissues versus normal colonic samples as well as worse prognosis in patients with high DIS3L2 expression. In addition, our RNA deep-sequencing data revealed that knockdown (KD) of DIS3L2 induces a strong transcriptomic disturbance in SW480 CRC cells. Moreover, gene ontology (GO) analysis of significant upregulated transcripts displays enrichment in mRNAs encoding proteins involved in cell cycle regulation and cancer-related pathways, which guided us to evaluate which specific hallmarks of cancer are differentially regulated by DIS3L2. To do so, we employed four CRC cell lines (HCT116, SW480, Caco-2 and HT-29) differing in their mutational background and oncogenicity. We demonstrate that depletion of DIS3L2 results in reduced cell viability of highly oncogenic SW480 and HCT116 CRC cells, but had little or no impact in the more differentiated Caco-2 and HT-29 cells. Remarkably, the mTOR signaling pathway, crucial for cell survival and growth, is downregulated after DIS3L2 KD, whereas AZGP1, an mTOR pathway inhibitor, is upregulated. Furthermore, our results indicate that depletion of DIS3L2 disturbs metastasis-associated properties, such as cell migration and invasion, only in highly oncogenic CRC cells. Our work reveals for the first time a role for DIS3L2 in sustaining CRC cell proliferation and provides evidence that this ribonuclease is required to support the viability and invasive behavior of dedifferentiated CRC cells. CI - (c) 2023. The Author(s). FAU - Garcia-Moreno, Juan F AU - Garcia-Moreno JF AD - Departamento de Genetica Humana, Instituto Nacional de Saude Doutor Ricardo Jorge, 1649-016, Lisbon, Portugal. AD - Faculdade de Ciencias, BioISI - Instituto de Biossistemas e Ciencias Integrativas, Universidade de Lisboa, 1749-016, Lisbon, Portugal. FAU - Lacerda, Rafaela AU - Lacerda R AD - Departamento de Genetica Humana, Instituto Nacional de Saude Doutor Ricardo Jorge, 1649-016, Lisbon, Portugal. AD - Faculdade de Ciencias, BioISI - Instituto de Biossistemas e Ciencias Integrativas, Universidade de Lisboa, 1749-016, Lisbon, Portugal. FAU - da Costa, Paulo J AU - da Costa PJ AD - Departamento de Genetica Humana, Instituto Nacional de Saude Doutor Ricardo Jorge, 1649-016, Lisbon, Portugal. AD - Faculdade de Ciencias, BioISI - Instituto de Biossistemas e Ciencias Integrativas, Universidade de Lisboa, 1749-016, Lisbon, Portugal. FAU - Pereira, Marcelo AU - Pereira M AD - Faculdade de Ciencias, BioISI - Instituto de Biossistemas e Ciencias Integrativas, Universidade de Lisboa, 1749-016, Lisbon, Portugal. FAU - Gama-Carvalho, Margarida AU - Gama-Carvalho M AD - Faculdade de Ciencias, BioISI - Instituto de Biossistemas e Ciencias Integrativas, Universidade de Lisboa, 1749-016, Lisbon, Portugal. FAU - Matos, Paulo AU - Matos P AD - Departamento de Genetica Humana, Instituto Nacional de Saude Doutor Ricardo Jorge, 1649-016, Lisbon, Portugal. AD - Faculdade de Ciencias, BioISI - Instituto de Biossistemas e Ciencias Integrativas, Universidade de Lisboa, 1749-016, Lisbon, Portugal. FAU - Romao, Luisa AU - Romao L AUID- ORCID: 0000-0002-5061-5287 AD - Departamento de Genetica Humana, Instituto Nacional de Saude Doutor Ricardo Jorge, 1649-016, Lisbon, Portugal. luisa.romao@insa.min-saude.pt. AD - Faculdade de Ciencias, BioISI - Instituto de Biossistemas e Ciencias Integrativas, Universidade de Lisboa, 1749-016, Lisbon, Portugal. luisa.romao@insa.min-saude.pt. LA - eng GR - UID/MULTI/04046/2019/Fundacao para a Ciencia e a Tecnologia/ GR - SFRH/BD/52495/2014/Fundacao para a Ciencia e a Tecnologia/ GR - PD/BD/142898/2018/Fundacao para a Ciencia e a Tecnologia/ PT - Journal Article DEP - 20230620 PL - Switzerland TA - Cell Mol Life Sci JT - Cellular and molecular life sciences : CMLS JID - 9705402 RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) RN - 0 (RNA, Messenger) RN - EC 3.1.- (Ribonucleases) RN - EC 3.1.- (DIS3L2 protein, human) RN - EC 3.1.- (Exoribonucleases) SB - IM MH - Humans MH - Caco-2 Cells MH - Cell Line, Tumor MH - *Colorectal Neoplasms/genetics/metabolism MH - Signal Transduction/genetics MH - TOR Serine-Threonine Kinases/genetics/metabolism MH - HCT116 Cells MH - Cell Proliferation/genetics MH - RNA, Messenger MH - Cell Movement/genetics MH - Ribonucleases/genetics MH - Gene Expression Regulation, Neoplastic MH - Exoribonucleases/genetics/metabolism PMC - PMC10282049 OTO - NOTNLM OT - AZGP1 OT - Cell migration OT - Cell viability OT - Colorectal cancer OT - DIS3L2 OT - mTOR COIS- The authors declare that they have no conflict of interest. EDAT- 2023/06/21 01:07 MHDA- 2023/06/22 06:42 PMCR- 2023/06/20 CRDT- 2023/06/20 23:33 PHST- 2023/02/06 00:00 [received] PHST- 2023/06/05 00:00 [accepted] PHST- 2023/05/12 00:00 [revised] PHST- 2023/06/22 06:42 [medline] PHST- 2023/06/21 01:07 [pubmed] PHST- 2023/06/20 23:33 [entrez] PHST- 2023/06/20 00:00 [pmc-release] AID - 10.1007/s00018-023-04833-5 [pii] AID - 4833 [pii] AID - 10.1007/s00018-023-04833-5 [doi] PST - epublish SO - Cell Mol Life Sci. 2023 Jun 20;80(7):185. doi: 10.1007/s00018-023-04833-5.