[Mechanism of hsa-miR-302a-3p-targeted VEGFA in the Inhibition of Proliferation of Gastric Cancer Cell].
Journal: 2019/April - Journal of Sichuan University (Medical Science Edition)
ISSN: 1672-173X
PUBMED: 31037899
Abstract:
To explore the role mechanism of hsa-miR-302a-3p overexpression in the inhibition of proliferation of gastric cancer cell SGC-7901 by targeted-regulating vascular endothelial growth factor A (VEGFA).

METHODS
The cell transfection was used to transfect hsa-miR-302a-3p mimic into miR mimic group and transfect pc-VEGFA into VEGFA group, and the two genes were co-transfected into miR+VEGFA group. The transfection efficiency was detected by RT-PCR and Western blot. The bioinformatics targeting prediction and fluorescein assay were used to verify the targeting relationship between the two genes. Cell proliferation was detected by CCK-8 test, and Transwell assay was used to detect the invasion ability of each group, and scratch assay was used to detect the migration ability of each group. The morphology changes of epithelial-mesenchymal transition (EMT) in cells were observed under microscope. Western blot was used to detect the protein expression levels of survival-related proteins Ki67 and Caspase-3, EMT-related proteins E-cadherin, Vimentin, N-cadherin and Snail and VEGFA downstream target genes p-P38, p-MAPKAPK and p-Hsp27.

RESULTS
VEGFA was the predicted target site of miR-302a-3p. Compared with control group, the number of cells, the invasion and migration rates were also reduced ( P<0.05) in miR mimic group, and the number of cells was increased ( P<0.05) as well as the invasion and migration rates in VEGFA group. Compared with VEGFA group, the number of cells, the invasion and migration rates were also decreased ( P<0.05) in miR+VEGFA group. The protein expression level of E-cadherin was up-regulated ( P<0.05) while the protein expression levels of Vimentin, N-cadherin and Snail were down-regulated ( P<0.05), and the protein expression levels of p-P38, p-MAPKAPK and p-Hsp27 were also down-regulated ( P<0.05).

CONCLUSION
hsa-miR-302a-3p overexpression can inhibit the proliferation and promote apoptosis of gastric cancer cell SGC-7901 by targeting negative regulation of VEGFA expression.

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