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Effect of Hsa-miR-411-3P on Gastric Cancer Cells and Related Mechanisms |
CHEN Xue-yan1,ZHANG Na2,CHEN Juan2,YANG Yan-hong3,ZHANG Ju-feng2,**() |
1 School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou 510006, China 2 School of Life Science and Biopharmaceutics, Guangdong Pharmaceutical University, Guangzhou 510006, China 3 The First Affiliated Hospital, School of Clinical Medicine, Guangdong Pharmaceutical University, Guangzhou 510080, China |
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Abstract Objective: To study the functional effects of Hsa-miR-411-3P on gastric cancer cells and related molecular mechanisms. Method: The effect of Hsa-miR-411-3P on the proliferation of gastric cancer cells SGC-7901 and AGS was detected by MTT; The effect of Hsa-miR-411-3P on the cycle and apoptosis of gastric cancer cells SGC-7901、 AGS was detected by flow cytometry; Three online tools: RNAhybrid2.1.2, Miranda3.3a, TargetScan7.0 predicted Hsa-miR-411-3P target genes, and combined with mRNA sequencing results, the intersection genes were considered reliable target genes, and then performed GO, KEGG analysis; Real-time PCR was used to detect the expression of Hsa-miR-411-3P target genes VAV3, ROCK2, PLD1, and PTCH1. Result: Overexpression of Hsa-miR-411-3P inhibited the proliferation of gastric cancer cells SGC-7901 and AGS, promoted the apoptosis of gastric cancer cells SGC-7901, AGS, arrested the cell cycle of SGC-7901 in S phase, and arrested AGS cell cycle in G1 phase. Three online tools: RNAhybrid2.1.2, Miranda3.3a, TargetScan7.0, predicted Hsa-miR-411-3P target genes, and combined with mRNA sequencing results, the intersection genes were considered to be reliable target genes, and a total of 235 intersection targets were obtained. Take 235 intersection target genes as target gene sets for subsequent analysis. GO and KEGG analysis of 235 cross-target genes found that their molecular function was concentrated in enzyme binding, protein binding, transferase activity, etc. Biological processes was concentrated in metabolic processes, assembly of cellular components, development of anatomical structures, and biogenesis of cellular components, etc.(P<0.05). KEGG signaling pathway was concentrated in insulin signaling pathway, cAMP signaling pathway, AMPK signaling pathway, FOXO signaling pathway, etc. (P<0.05). Insulin signaling pathway, AMPK signaling pathway, FOXO signaling pathway, and cAMP signaling pathway are all closely related to gastric cancer. The Hsa-miR-411-3P target genes VAV3、 ROCK2、 PLD1 and PTCH1, which are highly correlated with gastric cancer, all participate in the cAMP signaling pathway.VAV3、 ROCK2、 PLD1 and PTCH1 may have a negative regulatory relationship with Hsa-miR-411-3P, because in the sequencing results, the expression of Hsa-miR-411-3P was up-regulated after berberine treatment of SGC-7901 cells, but the expression of VAV3、 ROCK2、 PLD1 and PTCH1 was down-regulated. For this, verification was performed by using Real-time PCR. It was found that overexpression of Hsa-miR-411-3P would decrease the expression of VAV3 and ROCK2 mRNA in SGC-7901、 AGS cells; overexpression of Hsa-miR-411-3P would increase the expression of PLD1 mRNA in SGC-7901 cells and decrease the expression of PLD1 mRNA in AGS cells;overexpression of Hsa-miR-411-3P would decrease the expression of PTCH1 mRNA in SGC-7901 cells and increase the expression of PTCH1 mRNA in AGS cells. It was indicated that overexpression of Hsa-miR-411-3P could down-regulate the expression of target genes VAV3 and ROCK2, because the expression of PLD1 and PTCH1 was differently in two gastric cancer cells, which was controversial and need to be further studied by other gastric cancer cells. Conclusion: Overexpression of Hsa-miR-411-3P will decrease the expression of target genes VAV3、 ROCK2 to affect cAMP signaling pathway, inhibit the proliferation of SGC-7901, AGS cells, promote apoptosis, and arrest SGC-7901 cell cycle in S phase, AGS cell cycle in G1 phase.
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Received: 02 September 2019
Published: 18 May 2020
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Corresponding Authors:
Ju-feng ZHANG
E-mail: jfzhang111@163.com
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