Knockdown of Cbp/P300-interacting transactivator with Glu/Asp-rich carboxy-terminal domain 2 inhibits cell division and increases apoptosis in gastric cancer.

Abstract

BACKGROUND:

Cbp/P300-interacting transactivator with Glu/Asp-rich carboxy-terminal domain 2 (CITED2) is a pleiotropic protein associated with numerous cell functions, including transcription and differentiation. The role of CITED2 has been investigated in a number of malignancies; however, the roles of this protein in gastric cancers remain unclear. Therefore, we determined the role of CITED2 in gastric cancers.

MATERIALS AND METHODS:

Gastric cancer cell lines (MKN74, MKN28, 7901, and AGS) were used to assess CITED2 transcript levels. Messenger RNA levels were determined using quantitative polymerase chain reaction. Lentiviral vectors containing CITED2 small interfering RNA were used to knockdown CITED2 expression. Cell proliferation was assessed with fluorescent imaging and 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide assays. Apoptosis and cell cycle stages were assessed through flow cytometry, and formation of colonies was determined using a fluorescent microscope.

RESULTS:

All cell lines tested in this study expressed CITED2. The cell line expressing the highest levels of CITED2 (MKN74) showed significant knockdown of endogenous CITED2 expression on lentiviral infection. Cell proliferation was shown to be lower in CITED2 knockdown MKN74 cells. G1/S-phase cell cycle arrest was observed on silencing of CITED2 in MKN74 cells. A significant increase in apoptosis was observed on CITED2 knock down in MKN74 cells, while colony forming ability was significantly inhibited after knock down of CITED2.

CONCLUSIONS:

CITED2 supports gastric cancer cell colony formation and proliferation while inhibiting apoptosis making it a potential gene therapy target for gastric cancer.

• Authors: Ze Tang, Gan He, Jie Xu, Li Zhongfu
• Journal: The Journal of surgical research (J Surg Res) Vol. 211 Pg. 1-7 (05 01 2017) ISSN: 1095-8673 [Electronic] United States
• PMID: 28501104 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2016 Elsevier Inc. All rights reserved.

Chemical References

• Biomarkers, Tumor
• CITED2 protein, human
• Repressor Proteins
• Trans-Activators

Topics

• Apoptosis (genetics, physiology)
• Biomarkers, Tumor (genetics, metabolism)
• Cell Cycle (physiology)
• Cell Line, Tumor
• Cell Proliferation (genetics, physiology)
• Flow Cytometry
• Gene Expression Regulation, Neoplastic
• Gene Knockdown Techniques
• Humans
• Repressor Proteins (genetics, metabolism)
• Stomach Neoplasms (genetics, metabolism)
• Trans-Activators (genetics, metabolism)
• Tumor Stem Cell Assay