|Table of Contents|

Effect of down-regulating MACC1 by shRNA on invasion and migration of gastric carcinoma cells

Journal Of Modern Oncology[ISSN:1672-4992/CN:61-1415/R]

Issue:
2017 02
Page:
174-178
Research Field:
Publishing date:

Info

Title:
Effect of down-regulating MACC1 by shRNA on invasion and migration of gastric carcinoma cells
Author(s):
Yuan Yuan1; Zhu Zhengqiu2
1.School of Postgraduate,Xuzhou Medical University，Jiangsu Xuzhou 221000,China；2.Department of Oncology,the Affiliated Hospital of Xuzhou Medical University,Jiangsu Xuzhou 221000,China.
Keywords:
hepatic stellate cells; MACC1; hepatocyte growth factor; MMPs; grastric carcinoma; migration; invasion
PACS:
R735.2
DOI:
10.3969/j.issn.1672-4992.2017.02.003
Abstract:
Objective:By in vitro experiment to explore the regulatory effect of MACC1 gene silencing in HSC expressα-SMA,MMP-2,MMP-9 and HGF,and its effects on ability of migration and invasion of co-cultured gastric carcinoma cells.Methods:To construct targeting MACC1 gene eukaryotic expression interference plasmid (MACC1-shRNA) and non-specific irrelevant interference plasmid (shNC)，transient transfect them into HSC with lipofectamine-2000，named low-expression group(shRNA) and negative control group (NC) accordingly,non-transfected HSC treated as a blank control group(Blank-Ctrl).The above three groups of cells were cultured in accordance with routinel,the expression levels of MACC1,α-SMA,HGF,MMP-2 and MMP-9 in human hepatic stellate cells were detected by Western blot and RT-PCR.Three groups of cells were co-cultured with MGC803 gastric carcinoma cells in vitro,the capabilities of migration and invasion were measured by transwell assay.Results:Western-Blot results showed that the expression of MACC1,α-SMA,HGF,MMP-2 and MMP-9 protein in HSC was significantly lower than that in the negative group and blank control group after transfection of MACC1-shRNA (P＜0.05).Real-time PCR results showed that the expression levels of mRNA of MACC1,α-SMA,HGF,MMP-2 and MMP-9 in low-expression group HSC significantly decreased (P＜0.01),compared with the others groups.Transwell test indicated that the number of cells through membrane in low-expression groups were significantly less than the negatiave groups and blank control groups in MGC803 cell (P＜0.01).There had no statistical significance between blank control group and negative group（P＞0.05）.Conclusion:Silencing MACC1 in hepatic stellate cells,can inhibit the expression of MMP-2,MMP-9 and HGF,and decrease the migration and invasion of gastric cancer cells in co-culture.

References:

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