Effect of Hsa-miR-411-3P on Gastric Cancer Cells and Related Mechanisms

doi:10.13523/j.cb.1909002

China Biotechnology

2020, Vol. 40

Issue (4): 1-9 DOI: 10.13523/j.cb.1909002

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-feng^2,^**(

)

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.

Key words： Gastric cancer miRNA Hsa-miR-411-3P

Received: 02 September 2019 Published: 18 May 2020

ZTFLH:

R737.9

Corresponding Authors: Ju-feng ZHANG E-mail: jfzhang111@163.com

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	Xue-yan CHEN
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	Yan-hong YANG
	Ju-feng ZHANG

Cite this article:

CHEN Xue-yan,ZHANG Na,CHEN Juan,YANG Yan-hong,ZHANG Ju-feng. Effect of Hsa-miR-411-3P on Gastric Cancer Cells and Related Mechanisms. China Biotechnology, 2020, 40(4): 1-9.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.1909002 OR https://manu60.magtech.com.cn/biotech/Y2020/V40/I4/1

Table 1 Primer sequences

Table 2 Effect of Hsa-miR-411-3P on proliferation of gastric cancer cells

Fig.1 Effect of Hsa-miR-411-3P on proliferation of gastric cancer cells (a) Effect of Hsa-miR-411-3P on the growth of SGC-7901 cells (b) Effect of Hsa-miR-411-3P on the growth of AGS cells * P<0.05;** P<0.01;*** P<0.001

Fig.2 Effect of Hsa-miR-411-3P on cell cycle of gastric cancer (a)、(b) Cycle distribution of mimic NC group and Hsa-miR-411-3P mimic group in SGC-7901 cells, respectively (c)、(d) Cycle distribution of mimic NC group and Hsa-miR-411-3P mimic group in AGS cells, respectively (e) S-phase proportional histogram of mimic NC group and Hsa-miR-411-3P mimic group in SGC-7901 cells (f) G1 proportional histogram of mimic NC group and Hsa-miR-411-3P mimic group in AGS cells

Fig.3 Effect of Hsa-miR-411-3P on apoptosis of gastric cancer cells (a)(b) Apoptotic profiles of Mimic NC group and Hsa-miR-411-3P Mimic group in SGC-7901 cells, respectively (c)(d) Apoptotic profiles of Mimic NC group and Hsa-miR-411-3P Mimic group in AGS cells, respectively (e) Apoptotic rate histogram of Mimic NC group and Hsa-miR-411-3P Mimic group in gastric cancer cells SGC-7901, AGS * P<0.05;** P<0.01;*** P<0.001

Table 3 Functional analysis of GO molecules of Hsa-miR-411-3P target genes

Table 4 Analysis of GO biological process of Hsa-miR-411-3P target genes

Table 5 KEGG analysis of Hsa-miR-411-3P target genes

Fig.4 Hsa-miR-411-3P and its target genes VAV3, ROCK2, PLD1, PTCH1 expression in sequencing results (a)The expression of Hsa-miR-411-3P in sequencing results (b)The expression of Hsa-miR-411-3P target genes VAV3, ROCK2, PLD1, PTCH1 in sequencing results

Fig.5 Effect of Hsa-miR-411-3P on target gene expression (a) Effect of Hsa-miR-411-3P on the expression of target genes VAV3 and ROCK2 (b) Effect of Hsa-miR-411-3P on the expression of target genes PLD1 and PTCH1 * P<0.05;** P<0.01;*** P<0.001


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