Notch2 regulates matrix metallopeptidase 9 via PI3K/AKT signaling in human gastric carcinoma cell MKN-45

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外文摘要：AIM:To clarify the role of activated Notch2 in the invasiveness of gastric cancer.METHODS:To investigate the invasiveness of silencing Notch2 gene expression,we established a Notch2 small interfering RNA (siRNA) transfected cell line using the MKN-45 gastric cancer cell line.After the successful transfection confirmed by real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting,migration and invasion assays were employed to evaluate the aggressiveness of the gastric cancer.RT-PCR and Western blottings were employed to confirm the down-regulation of Notch2 and to evaluate the expression of epithelial mesenchymal transition-related gene matrix metallopeptidase 9 (MMPg),Akt,p-Akt.To confirm the relationship between PI3K-Akt and MMP9,the PI3K inhibitor LY294002 was used to treat MKN-45 cells.RESULTS:Notch2 expression was dramatically decreased after Notch2 siRNA transfection (100.00％ ±9.74％ vs 11.61％ ± 3.85％,P ＜ 0.01 by qRT-PCR).There was also a marked reduction of Notch target gene Hes1 (100.00％ ± 4.74％ vs 61.61％ ± 3.58％,P ＜ 0.05)at the mRNA,indicating an inhibition of Notch signaling.Inhibition of Notch signaling was also confirmed by the marked reduction of Notch2 intracellular domain at the protein levels (100.00％ ± 9.74％ vs 65.61％ ± 7.58％,P ＜ 0.05).Down-regulation of Notch2 by siRNA enhanced tumor cell invasion (100.00％ ± 21.64％ vs 162.22％ ± 16.84％,P ＜ 0.05) and expression of MMP9 (1.56 fold,P ＜ 0.05),and activated the pro-MMP9 protein to its active form (1.48 fold,P ＜ 0.05).There was no significant difference in the protein levels of Akt between the two groups (100.00％ ± 10.87％ vs 96.61％ ± 7.33％,P ＞ 0.05),while down-regulation of Notch2 elevated p-Akt expression (100.00％ ± 9.87％ vs 154.61％ ± 13.10％,P ＜ 0.05).Furthermore,p-Akt and MMP9 was down-regulated in response to the inhibitor LY294002 (p-Akt 100.00％ ± 8.87％ vs 58.27％ ± 5.01％,P ＜ 0.05; MMP9 100.00％ ± 9.17％ vs 50.03％ ± 4.88％,P ＜ 0.05).CONCLUSION:Notch2 may negatively regulate cell invasion by inhibiting the PI3K-Akt signaling pathway in gastric cancer.

外文关键词：

Notch2StomachCancerInvasionEpithelial mesenchymal transitionMatrix metallopeptidase 9RNA interference

作者：

Ling-Yun Guo、Yu-Min Li、Liang Qiao、Tao Liu、Yuan-Yuan Du、Jun-Qiang Zhang、Wen-Ting He

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作者单位：

the Second Clinical Medical School of Lanzhou University, the Second Hospital of Lanzhou University, Lanzhou 730030, Gansu Province, China

Key Laboratory of Digestive System Tumors, Department of General Surgery, the Second Hospital of Lanzhou University, Lanzhou 730030, Gansu Province, China

Storr Liver Unit at the Westmead Millennium Institute, the University of Sydney at Westmead Hospital, Westmead, NSW 2145, Australia

Key Laboratory of Digestive System Tumors, the Second Hospital of Lanzhou University, Lanzhou 730030, Gansu Province, China

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基金：

国家自然科学基金Fund for Key Laboratory of Digestive System Tumors,Gansu Province

项目编号：

30870364lzujbky-2011-t03

出版年：

2012

DOI：

10.3748/wjg.v18.i48.7262

世界胃肠病学杂志(英文版)