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©The Author(s) 2018. Published by Baishideng Publishing Group Inc. All rights reserved.
Piwi like RNA-mediated gene silencing 1 gene as a possible major player in gastric cancer
Taíssa Araújo, André Khayat, Luciana Quintana, Danielle Calcagno, Ronald Mourão, Antônio Modesto, Juliana Paiva, Adhara Lima, Fabiano Moreira, Edivaldo Oliveira, Michel Souza, Moneeb Othman, Thomas Liehr, Eliana Abdelhay, Renata Gomes, Sidney Santos, Paulo Assumpção
Taíssa Araújo, André Khayat, Luciana Quintana, Danielle Calcagno, Ronald Mourão, Antônio Modesto, Juliana Paiva, Adhara Lima, Fabiano Moreira, Sidney Santos, Paulo Assumpção, Núcleo de Pesquisas em Oncologia, Universidade Federal do Pará, Belém 66073-000, Brazil
Edivaldo Oliveira, Michel Souza, Laboratório de Cultura de Tecidos e Citogenética, Instituto Evandro Chagas, Belém 66087-082, Brazil
Moneeb Othman, Thomas Liehr, Institute of Human Genetics, Universitätsklinikum Jena, Jena 07747, Germany
Eliana Abdelhay, Renata Gomes, Laboratório de Célula Tronco, Centro de Transplante de Medula Óssea, Instituto Nacional de Câncer José Alencar Gomes da Silva, Rio de Janeiro 20230-130, Brazil
Author contributions: Araújo T and Khayat A contributed equally to this work; Araújo T and Khayat A performed the majority of experiments; Moreira F, Oliveira E and Souza M analyzed the data; Modesto A, Paiva J and Lima A performed the molecular investigations; Quintana L, Calcagno D and Mourão R participated equally in cell culture management; Othman M and Liehr T performed molecular cytogenetics experiments; Abdelhay E and Gomes R performed proteomic assay; Santos S and Assumpção P designed and coordinated the research; Araújo T , Khayat A and Quintana L wrote the paper.
Supported by Fundação Amazônia de Amparo a Estudos e Pesquisa (FAPESPA), No. 174/2014.
Conflict-of-interest statement: The authors declare that they have no conflicts of interest.
Data sharing statement: No additional data are available.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Corresponding author to: Paulo Assumpção, MD, MSc, PhD, Academic Research, Adjunct Professor, Surgical Oncologist, Núcleo de Pesquisas em Oncologia, Universidade Federal do Pará, Rua dos Mundurucus 4487, Belém 66073-000, Brazil. assumpcaopp@gmail.com
Telephone: +55-91-984171112
Received: August 14, 2018
Peer-review started: August 14, 2018
First decision: August 31, 2018
Revised: September 7, 2018
Accepted: October 5, 2018
Article in press: October 5, 2018
Published online: December 21, 2018
AIM
To establish a permanent piwi like RNA-mediated gene silencing 1 (PIWIL1) gene knockout in AGP01 gastric cancer cell line using CRISPR-Cas9 system and analyze phenotypic modifications as well as gene expression alterations.
METHODS
CRISPR-Cas9 system used was purchased from Dharmacon GE Life Sciences (Lafayette, CO, United States) and permanent knockout was performed according to manufacturer’s recommendations. Wound-healing assay was performed to investigate the effect of PIWIL1 knockout on migration capability of cells and Boyden chamber invasion assay was performed to investigate the effect on invasion capability. For the gene expression analysis, a one-color microarray-based gene expression analysis kit (Agilent Technologies, Santa Clara, CA, United States) was used according to the protocol provided by the manufacturer.
RESULTS
PIWIL1 gene knockout caused a significant decrease in AGP01 migration capacity as well as a significant decrease in cell invasiveness. Moreover, functional analysis based on grouping of all differentially expressed mRNAs identified a total of 35 genes (5 up-regulated and 30 down-regulated) encoding proteins involved in cellular invasion and migration. According to current literature, 9 of these 35 genes (DOCK2, ZNF503, PDE4D, ABL1, ABL2, LPAR1, SMAD2, WASF3 and DACH1) are possibly related to the mechanisms used by PIWIL1 to promote carcinogenic effects related to migration and invasion, since their functions are consistent with the changes observed (being up- or down-regulated after knockout).
CONCLUSION
Taken together, these data reinforce the idea that PIWIL1 plays a crucial role in the signaling pathway of gastric cancer, regulating several genes involved in migration and invasion processes; therefore, its use as a therapeutic target may generate promising results in the treatment of gastric cancer.
Core tip:Piwi like RNA-mediated gene silencing 1 (PIWIL1) gene emerged as an interesting target for gastric cancer, as it is expressed in cancer, stem and germ cells, but it is absent in normal somatic tissue. Our results propose that PIWIL1 plays a crucial role in the signaling pathway of gastric cancer, regulating several genes involved in migration and invasion processes; therefore, its use as a therapeutic target may generate promising results in the treatment of gastric cancer.
