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Published on: April 28, 2021
ARID1A is involved in DNA double-strand break repair in gastric cancer
Ying Zhang1, He-Sheng Qian1, Gengwei Hu2
1Department of Oncology, Fuyang Cancer Hospital, Fuyang, China.
Background:
Defects in DNA damage repair can cause genetic mutations, which in turn can cause different types of cancers. Chromatin remodeling complexes, which help repair damaged DNA, can cause the chromatin structure to change as a result of DNA damage. ARID1A may play a role in the process of DNA damage repair, and arid1a may be related to the occurrence and development of gastric cancer (GC). This study aimed to investigate the mechanism of ARID1A regulating the DNA damage repair of gastric adenocarcinoma cell lines AGS and SGC-7901 and its effect on migration, proliferation and apoptosis.
Methods:
The expression of ARID1A plasmid was detected by Western blot and real-time polymerase chain reaction (PCR). The effect of etoposide (ETO) on the survival rate of AGS and SGC-7901 gastric adenocarcinoma cell lines was detected by MTT assay. The DNA double-strand break model was established by ETO and then passed through the comet assay and immunofluorescence co-localization to observe DNA damage; western blot method was used to detect the effect of ARID1A on the expression of related proteins in DNA damage repair pathway in gastric adenocarcinoma cells; scratch test and colony formation experiments were used to observe ARID1A migration and proliferation of gastric adenocarcinoma cells. The flow cytometry was used to detect the effect of ARID1A on apoptosis of gastric adenocarcinoma cells.
Results:
The expression of mRNA and protein was increased after transfection of ARID1A plasmid. ETO was confirmed by MTT assay to inhibit cell survival in a dose-dependent manner. After the DNA double-strand break model was established by ETO, the expression levels of phospho-ataxia telangiectasia mutated (p-ATM) protein increased in the overexpressed ARID1A group. Meanwhile, the overexpressed ARID1A group had a shortened tail moment, and γ-H2AX and ARID1A co-localized in the DNA damage site of the nucleus. The over-expressed ARID1A group had weaker wound healing ability, reduced number of clone formation, and increased apoptosis rate.
Conclusions:
ARID1A may repair DNA double-strand breaks caused by ETO by p-ATM pathway; ARID1A can inhibit the migration and proliferation of gastric adenocarcinoma cells and promote apoptosis. Our findings indicate that ARID1A could serve as a therapeutic target and biomarker for GC patients.
Insights
The study shows that ARID1A helps repair DNA double-strand breaks in gastric cancer cells. Overexpressing ARID1A inhibits cancer cell migration and proliferation while promoting apoptosis, suggesting it as a potential therapeutic target.
Area of Science:
- Molecular Biology
- Cancer Research
- Genetics
Background:
- Defects in DNA damage repair are linked to genetic mutations and cancer development.
- Chromatin remodeling complexes, including ARID1A, play a role in DNA repair and may influence gastric cancer (GC) progression.
- This study investigates ARID1A's mechanism in DNA damage repair within gastric adenocarcinoma cells.
Purpose of the Study:
- To elucidate the mechanism by which ARID1A regulates DNA damage repair in gastric adenocarcinoma cell lines (AGS and SGC-7901).
- To assess the impact of ARID1A on the migration, proliferation, and apoptosis of these cancer cells.
Main Methods:
- ARID1A expression was analyzed using Western blot and real-time PCR.
- Etoposide (ETO) was used to induce DNA double-strand breaks, with cell viability assessed by MTT assay.
- DNA damage, protein expression (p-ATM, γ-H2AX), cell migration (scratch test), proliferation (colony formation), and apoptosis (flow cytometry) were evaluated.
Main Results:
- ARID1A overexpression increased mRNA and protein levels and enhanced p-ATM expression following ETO treatment.
- ARID1A co-localized with γ-H2AX at DNA damage sites and reduced DNA damage indicators (tail moment).
- Overexpression of ARID1A inhibited cell migration and proliferation while increasing apoptosis rates.
Conclusions:
- ARID1A appears to repair ETO-induced DNA double-strand breaks via the p-ATM pathway.
- ARID1A demonstrates inhibitory effects on gastric adenocarcinoma cell migration and proliferation and promotes apoptosis.
- ARID1A presents potential as a therapeutic target and biomarker for GC patients.
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