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  • miR‑494‑BAG‑1 axis is involved in cinobufacini‑induced cell proliferation and apoptosis in gastric cancer.

miR‑494‑BAG‑1 axis is involved in cinobufacini‑induced cell proliferation and apoptosis in gastric cancer.

Molecular medicine reports (2018-03-24)
Zhili Shen, Yan Li, Chengcheng Zhao, Fen Wang, Rongping Zhou, Gang Chen
ABSTRACT

Cinobufacini is widely used in the treatment of advanced cancers. It has been previously reported that microRNA (miR)‑494 was upregulated in cinobufacini‑treated gastric cancer cells; however, the detailed role of miR‑494 in the anti‑tumor activity of cinobufacini is unclear. The present study aimed to clarify the function of miR‑494 in cinobufacini‑induced cell behavior changes. Cell viability and proliferation ability were investigated using a Cell Counting Kit‑8 assay. Flow cytometry was performed to investigate the apoptosis rate of gastric cancer (GC) cells. The mRNA expression levels of microRNA (miR)‑494 and BCL2 associated athanogene 1 (BAG‑1) were investigated using reverse transcription‑quantitative polymerase chain reaction, and the protein expression level of BAG‑1 was investigated using western blot assays. The results demonstrated that treatment with cinobufacini suppressed proliferation and promoted apoptosis of gastric cancer cells. miR‑494 acts as a tumor suppressor gene in gastric cancer. In cinobufacini‑treated cells, miR‑494 and BAG‑1 exhibited opposing expression trends. Furthermore, knockdown of miR‑494 in cinobufacini‑treated cells upregulated the protein expression level of BAG‑1, promoted cell proliferation and inhibited cell apoptosis. In addition, inhibition of BAG‑1 using small interfering RNA in cinobufacini‑treated cells partially abrogated the effects of miR‑494 inhibitor on cell proliferation and apoptosis. Thus, these results suggest that cinobufacini suppresses GC cells proliferation and promotes apoptosis partially through the regulation of miR‑494‑BAG‑1 axis, which may provide a novel insight into the functional mechanism of cinobufacini.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
(Tyr[SO3H]27)Cholecystokinin fragment 26-33 Amide, ≥97% (HPLC), powder
Sigma-Aldrich
Monoclonal Anti-β-Actin antibody produced in mouse, clone AC-15, ascites fluid