Mechanism of hsa-miR-302a-3p-targeted VEGFA in the Inhibition of Proliferation of Gastric Cancer Cell

YANG Chun, DENG Shao-ping

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.

 

Keywords: hsa-miR-302a-3p, VEGFA, Gastric cancer cell 

 

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References


BOURKE В. Will treatment of Helicobacter pylori infection in childhood alter the risk of developing gastric cancer. Can J Gastroenterol ,2005,19(7); 409-411.

SCHWARZENBACH H, NISH IDA N, CALIN GA, et al. Clinical relevance of circulating cell-free microRNAs in cancer. Nat Rev Clin Oncol, 2014,11(3) : 145-156.

NOGUCHI S, YASUI Y, IWASAKI J. et al. Replacement treatment with microRNA-143 and -145 induces synergistic inhibition of the growth of human bladder cancer cells by regulating PI3K/Akt and МАРК signaling pathways. Cancer Lett,2013,328(2):353-361.

HU S. WILSON KD, GHOSH Z, et al. MicroRNA-302 increases reprogramming efficiency via repression of NR2F2. Stem Cells,2013,31 (2):259-268.

QINC, ZHA W. FAN R, et at. MicroRNA-302a inhibits cell proliferation and invasion, and induces cell apoptosis in hepatocellular carcinoma by directly targeting VEGFA. Mol Med Rep,2017,16(5):6360-6367.

GOEL HL, PURSELL B, SHULTZ LD, et al. P-Rexl promotes resistance to VEGF/VEGFR-targeted therapy in prostate cancer. Cell Rep,2016,14(9):2193-2208.

MACEDO F. LADEIRA K, LONGATTOFILHO A, et al. Gastric cancer and angiogenesis; is VEGF a useful biomarker to assess progression and remission? J Gastric Cancer, 2017, 17(1): 1-10.

PENG S, GAO Г), GAO C, et al. MicroRNAs regulate signaling pathways in osteogenic differentiation of mesenchymal stem cells (Review). Mol Med Rep. 2016, 14 (1):623-629.

HIBINOS' SAITO Y. MURAMATSU T, et al. Tul900 MiR-1246 and Mir-302 are novel targets of epigenetic therapy with dznep and SAHA in human cancer cells. Gastroenterology,2013,144(5) :S876-S876.

MA G, LI Q, DAI W, etal. Prognostic implications of miR- 302a/b/c/d in human gastric cancer. Pathol Oncol Res, 2017 , 23(4):899-905.

WANG X, CHEN X, FANG J, et al. Overexpression of both VEGF-A and VEGF-C in gastric cancer correlates with prognosis, and silencing of both is effective to inhibit cancer growth. Int J Exp Pathol,2013,6(4):586-597.

TAMMA R, ANNESE T, RUGGIERI S. et al. VEGFA and VEGFR2 RNA scope determination in gastric cancer. J Mol Histol,2018,49(4) :429-435.

BURGESSER MV, RIVA V, OJEDA SM, et al. Expression of VEGF-A, HIF-1 A, CD34 and Ki67 in clear cell renal cell carcinomas and their relationship with conventional prognostic markers. Rev Fac Cien Med Univ Nac Cordoba, 2014,71(1):7-15.

JAVADB, TAYEBE R, ADELEH D. et al. Induction of apoptosis by green synthesized gold nanoparticles through activation of caspase-3 and 9 in human cervical cancer cells. Avicenna J Med Biotechnol.2016,8(2):75-83.

MARKOSYAN N, RECH A, VONDERHEIDE RH, et al. Abstract 469; mPGESl deletion and the mechanisms of tumor growth suppression. Cancer Res, 2015, 75 ( 15 Supplement);469-469.

LAMOUILLE S, XU J. DERYNCK R. Molecular mechanisms of epithelial-mesenchymal transition. Nat Rev Mol Cell Biol,2014,15(3): 178-196.

XU M, ZHOU H, ZHANG C, et al. A DAM 17 promotes epithelial-mesenchymal transition via TGF-p/Smad pathway in gastric carcinoma cells. Int J Oncol, 2016 . 49 ( 6 ): 2520- 2528.

MASUI T. OTA I. YOOK JI. et al. Snail-induced epithelial-mesenchymal transition promotes cancer stem cell¬like phenotype in head and neck cancer cells. Int J Oncol, 2014 *44(3) :693-699.

DESAI P. YANG J, TIAN B, etui. Mixed-effects model of epithelial-mesenchymal transition reveals rewiring of signaling networks. Cell Signal .2015 ,27( 7): 1413-1425.

ZHANG Z, RUI W. WANG ZC, et al. Anti-proliferation and anti-metastasis effect of barbaloin in non-small cell lung cancer via inactivating p38MAPK/Cdc25B/Hsp27 pathway. Oncol Rep, 2017,38 (2): 1172-1180.

SAW AD A J. LI F, KOMATSU M. R-Ras inhibits VEGF- Induced p38MAPK activation and HSP27 phosphorylation in endothelial cells. J Vase Res,2015,52(5);347-359.


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