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Published on: May 21, 2019
MicroRNA-328 acts as an anti-oncogene by targeting ABCG2 in gastric carcinoma
1Department of Gastroenterology, Weifang People's Hospital, Weifang, China. anjunyan108@163.com.
Objective:
To explore the regulatory mechanism of microRNA-328 expression level by targeting the protein ATP Binding Cassette Transporter G2 (ABCG2) in gastric cancer cells and seek for a biological marker of predicting gastric cancer.
Patients And Methods:
SGC-7901 and MKN-28 human gastric cancer cell lines were cultured. Meanwhile, paired gastric cancer pathological tissues and the corresponding adjacent normal tissues were collected. Western blot analysis was used to validate the protein expression of ABCG2. Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) analysis was used to detect the mRNA expression level of miR-328 and ABCG2. Cell counting kit-8 (CCK-8) and colony formation assay were performed to validate the proliferous ability of human gastric cancer cells. The transwell invasion and migration were operated to determine the migratory and invasive capacity. Dual-Luciferase reporter assay, qRT-PCR and Western blot were used to prove the target of miR-328.
Results:
Bioinformatics analysis made a prediction that ABCG2 was a direct functional target of miR-328. Position 619-625 of ABCG2 3'-UTR had a space structure that was complementary to miR-328 by bioinformatics analysis, and there was a significant reduction in the level of miR-328 in human gastric cancer cell lines and tissues. The expression of miR-328 down-regulated proliferation, invasion and migration of human gastric cancer cells in vitro, while silencing of miR-328 accelerated proliferation, invasion and migration of human gastric cancer cells in vitro. All results displayed ABCG2 was direct target protein of miR-328 owing the binding site and they presented a negative correlation.
Conclusions:
ABCG2 is the target protein of miR-328. It presents a negative correlation of the expression level between miR-328 and ABCG2. Down-regulation of miR-328 inhibits the proliferation, invasion and migration of gastric cancer cell lines. MiR-328 could predict generation and development of gastric cancer as a biomarker.
Insights
MicroRNA-328 (miR-328) targets ATP Binding Cassette Transporter G2 (ABCG2) in gastric cancer, inhibiting its growth. This miR-328/ABCG2 interaction shows potential as a biomarker for gastric cancer development.
Area of Science:
- Molecular biology
- Cancer research
- Biomarker discovery
Background:
- Gastric cancer is a significant global health concern.
- Understanding the molecular mechanisms driving gastric cancer progression is crucial for developing effective treatments.
- MicroRNAs play vital roles in regulating gene expression and are implicated in various cancers.
Purpose of the Study:
- To investigate the regulatory relationship between microRNA-328 (miR-328) and ATP Binding Cassette Transporter G2 (ABCG2) in gastric cancer.
- To explore the potential of miR-328 as a biomarker for predicting gastric cancer.
- To elucidate the role of the miR-328/ABCG2 axis in gastric cancer cell proliferation, invasion, and migration.
Main Methods:
- Utilized human gastric cancer cell lines (SGC-7901, MKN-28) and patient tissues.
- Employed Western blot and quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) to assess protein and mRNA expression levels.
- Performed cell counting kit-8 (CCK-8), colony formation, and Transwell assays to evaluate cellular proliferation, migration, and invasion.
- Conducted Dual-Luciferase reporter assays to confirm the direct targeting of ABCG2 by miR-328.
Main Results:
- Bioinformatics predicted and experimental validation confirmed ABCG2 as a direct functional target of miR-328.
- A significant reduction in miR-328 levels was observed in gastric cancer cell lines and tissues.
- Down-regulation of miR-328 expression promoted proliferation, invasion, and migration of gastric cancer cells.
- A negative correlation was established between miR-328 and ABCG2 expression levels.
Conclusions:
- ABCG2 is a direct target protein of miR-328 in gastric cancer.
- The expression levels of miR-328 and ABCG2 are negatively correlated.
- Down-regulation of miR-328 significantly inhibits gastric cancer cell proliferation, invasion, and migration.
- MiR-328 holds promise as a potential biomarker for predicting the generation and development of gastric cancer.
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