去氢骆驼蓬碱通过PTEN/Akt/MDM2信号通路抑制胃癌细胞COX-2表达
|
摘要
目的探讨PTEN基因/Akt/小鼠双微体基因(MDM2)信号通路在去氢骆驼蓬碱(harmine,HM)抑制胃癌细胞COX-2表达中的作用。方法构建PTEN(PTEN-siRNA)、Akt(Akt-siRNA)和MDM2(MDM2-siRNA)小干扰RNA,分别转染胃癌细胞株SGC-7901和MKN-45,Western blot检测HM干预前后PTEN、p-Akt、p-MDM2和COX-2表达。结果 HM诱导PTEN表达,抑制Akt、MDM2磷酸化和COX-2表达。敲减PTEN基因有效地阻断了HM抑制Akt、MDM2磷酸化和COX-2表达的作用,敲减Akt基因可协同HM抑制MDM2磷酸化和COX-2表达,敲减MDM2基因可协同HM抑制COX-2表达。结论 HM可能通过PTEN/Akt/MDM2信号通路,下调胃癌细胞COX-2表达。
Aim To further analyze the effects of PTEN/Akt/MDM2 signaling pathway in the harmine(HM)-mediated inhibition of COX-2 expression in gastric cancer cells. Methods PTEN-siRNA,Akt-siRNA,MDM2-siRNA were constructed and respectively transfected into SGC-7901 and MKN-45 cells,and then added or not added HM for 24 h. The expression of PTEN,Akt and phosphorylated Akt(p-Akt),MDM2 and phosphorylated MDM2(p-MDM2),as well as COX-2 expression was detected by Western blot. Results HM increased PTEN expression,but inhibited p-Akt,p-MDM2 and COX-2 expression in SGC-7901 and MKN-45 cells. Knockdown of PTEN blocked HM-induced inhibition of Akt and MDM2 phosphorylation,as well as down-regulation of COX-2 expression. Knockdown of Akt and treatment with HM synergistically inhibited p-MDM2 and COX-2 expression. Knockdown of MDM2 and treatment with HM synergistically inhibited COX-2 protein expression. Conclusions HM down-regulates the expression of COX-2 protein in gastric cancer cells via PTEN/Akt/MDM2 signaling pathway.
Aim To further analyze the effects of PTEN/Akt/MDM2 signaling pathway in the harmine(HM)-mediated inhibition of COX-2 expression in gastric cancer cells. Methods PTEN-siRNA,Akt-siRNA,MDM2-siRNA were constructed and respectively transfected into SGC-7901 and MKN-45 cells,and then added or not added HM for 24 h. The expression of PTEN,Akt and phosphorylated Akt(p-Akt),MDM2 and phosphorylated MDM2(p-MDM2),as well as COX-2 expression was detected by Western blot. Results HM increased PTEN expression,but inhibited p-Akt,p-MDM2 and COX-2 expression in SGC-7901 and MKN-45 cells. Knockdown of PTEN blocked HM-induced inhibition of Akt and MDM2 phosphorylation,as well as down-regulation of COX-2 expression. Knockdown of Akt and treatment with HM synergistically inhibited p-MDM2 and COX-2 expression. Knockdown of MDM2 and treatment with HM synergistically inhibited COX-2 protein expression. Conclusions HM down-regulates the expression of COX-2 protein in gastric cancer cells via PTEN/Akt/MDM2 signaling pathway.
引文
[1] Li S,Wang A,Gu F,et al.Novel harmine derivatives for tumor targeted therapy [J].Oncotarget,2015,6(11):8988-9001.
[2] Zhang H,Sun K,Ding J,et al.Harmine induces apoptosis and inhibits tumor cell proliferation,migration and invasion through down-regulation of cyclooxygenase-2 expression in gastric cancer [J].Phytomedicine,2014,21(3):348-55.
[3] Jing X,Cheng W,Wang S,et al.Resveratrol induces cell cycle arrest in human gastric cancer MGC803 cells via the PTEN-regulated PI3K/Akt signaling pathway [J].Oncol Rep,2016,35(1):472-8.
[4] Zheng T,Meng X,Wang J,et al.PTEN- and p53-mediated apoptosis and cell cycle arrest by FTY720 in gastric cancer cells and nude mice [J].J Cell Biochem,2010,111(1):218-28.
[5] Sepideh S,Mohammadreza J N,Ali D,et al.Study of the murine double minute 2 status in patients with gastric and colorectal carcinomas and its correlation with prognostic factors [J].Indian J Pathol Microbiol,2012,55(2):192-5.
[6] Jung C R,Lim J H,Choi Y,et al.Enigma negatively regulates p53 through MDM2 and promotes tumor cell survival in mice [J].J Clin Invest,2010,120(12):4493-506.
[7] 周林云,任文艳,廖云鹏,等.IGFBP-5及p53与汉防己甲素抑制乳腺癌MCF-7细胞增殖的关系研究[J].中国药理学通报,2018,34(1):38-43.[7] Zhou L Y,Ren W Y,Liao Y P,et al.Study on relationship between IGFBP-5/p53 axis and anti-proliferation effect of tetrandrine in MCF-7 cells [J].Chin Pharmacol Bull,2018,34(1):38-43.
[8] He X P,Shao Y,Li X L,et al.Downregulation of miR-101 in gastric cancer correlates with cyclooxygenase-2 overexpression and tumor growth [J].FEBS J,2012,279(22):4201-12.
[9] 孙为豪,朱悦,周逸婵,等.熊果酸抑制胃癌细胞COX-2表达的信号转导研究[J].中国药理学通报,2016,32(7):925-32.[9] Sun W H,Zhu Y,Zhou Y C,et al.Signal transduction pathway of ursolic acid inhibiting COX-2 expression in gastric cancer cells [J].Chin Pharmacol Bull,2016,32(7):925-32.
[10] Sun W H,Zhu F,Chen G S,et al.Blockade of cholecystokinin-2 receptor and cyclooxygenase-2 synergistically induces cell apoptosis,and inhibits the proliferation of human gastric cancer cells in vitro [J].Cancer Lett,2008,263(2):302-11.
[11] Zhu Y,Dai B,Zhang H,et al.Long non-coding RNA LOC572558 inhibits bladder cancer cell proliferation and tumor growth by regulating the AKT-MDM2-p53 signaling axis [J].Cancer Lett,2016,380(2):369-74.
[12] Mayo L D,Donner D B.The PTEN,Mdm2,p53 tumor suppressor-oncoprotein network [J].Trends Biochem Sci,2002,27(9):462-7.
[13] Oliner J D,Saiki A Y,Caenepeel S.The role of MDM2 amplification and overexpression in tumorigenesis [J].Cold Spring Harb Perspect Med,2016,6(6).pii:a026336.
[14] Kuo H P,Chuang T C,Tsai S C,et al.Berberine,an isoquinoline alkaloid,inhibits the metastatic potential of breast cancer cells via Akt pathway modulation[J].J Agric Food Chem,2012,60(38):9649-58.
[15] Dung T D,Day C H,Binh T V,et al.PP2A mediates diosmin p53 activation to block HA22T cell proliferation and tumor growth in xenografted nude mice through PI3K-Akt-MDM2 signaling suppression[J].Food Chem Toxicol,2012,50(5):1802-10.
[2] Zhang H,Sun K,Ding J,et al.Harmine induces apoptosis and inhibits tumor cell proliferation,migration and invasion through down-regulation of cyclooxygenase-2 expression in gastric cancer [J].Phytomedicine,2014,21(3):348-55.
[3] Jing X,Cheng W,Wang S,et al.Resveratrol induces cell cycle arrest in human gastric cancer MGC803 cells via the PTEN-regulated PI3K/Akt signaling pathway [J].Oncol Rep,2016,35(1):472-8.
[4] Zheng T,Meng X,Wang J,et al.PTEN- and p53-mediated apoptosis and cell cycle arrest by FTY720 in gastric cancer cells and nude mice [J].J Cell Biochem,2010,111(1):218-28.
[5] Sepideh S,Mohammadreza J N,Ali D,et al.Study of the murine double minute 2 status in patients with gastric and colorectal carcinomas and its correlation with prognostic factors [J].Indian J Pathol Microbiol,2012,55(2):192-5.
[6] Jung C R,Lim J H,Choi Y,et al.Enigma negatively regulates p53 through MDM2 and promotes tumor cell survival in mice [J].J Clin Invest,2010,120(12):4493-506.
[7] 周林云,任文艳,廖云鹏,等.IGFBP-5及p53与汉防己甲素抑制乳腺癌MCF-7细胞增殖的关系研究[J].中国药理学通报,2018,34(1):38-43.[7] Zhou L Y,Ren W Y,Liao Y P,et al.Study on relationship between IGFBP-5/p53 axis and anti-proliferation effect of tetrandrine in MCF-7 cells [J].Chin Pharmacol Bull,2018,34(1):38-43.
[8] He X P,Shao Y,Li X L,et al.Downregulation of miR-101 in gastric cancer correlates with cyclooxygenase-2 overexpression and tumor growth [J].FEBS J,2012,279(22):4201-12.
[9] 孙为豪,朱悦,周逸婵,等.熊果酸抑制胃癌细胞COX-2表达的信号转导研究[J].中国药理学通报,2016,32(7):925-32.[9] Sun W H,Zhu Y,Zhou Y C,et al.Signal transduction pathway of ursolic acid inhibiting COX-2 expression in gastric cancer cells [J].Chin Pharmacol Bull,2016,32(7):925-32.
[10] Sun W H,Zhu F,Chen G S,et al.Blockade of cholecystokinin-2 receptor and cyclooxygenase-2 synergistically induces cell apoptosis,and inhibits the proliferation of human gastric cancer cells in vitro [J].Cancer Lett,2008,263(2):302-11.
[11] Zhu Y,Dai B,Zhang H,et al.Long non-coding RNA LOC572558 inhibits bladder cancer cell proliferation and tumor growth by regulating the AKT-MDM2-p53 signaling axis [J].Cancer Lett,2016,380(2):369-74.
[12] Mayo L D,Donner D B.The PTEN,Mdm2,p53 tumor suppressor-oncoprotein network [J].Trends Biochem Sci,2002,27(9):462-7.
[13] Oliner J D,Saiki A Y,Caenepeel S.The role of MDM2 amplification and overexpression in tumorigenesis [J].Cold Spring Harb Perspect Med,2016,6(6).pii:a026336.
[14] Kuo H P,Chuang T C,Tsai S C,et al.Berberine,an isoquinoline alkaloid,inhibits the metastatic potential of breast cancer cells via Akt pathway modulation[J].J Agric Food Chem,2012,60(38):9649-58.
[15] Dung T D,Day C H,Binh T V,et al.PP2A mediates diosmin p53 activation to block HA22T cell proliferation and tumor growth in xenografted nude mice through PI3K-Akt-MDM2 signaling suppression[J].Food Chem Toxicol,2012,50(5):1802-10.