白藜芦醇抑制人结肠癌细胞增殖与骨形态发生蛋白7的关系研究
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摘要
目的研究白藜芦醇(resveratrol,Res)对人结肠癌细胞生长的抑制作用与骨形态发生蛋白7(BMP7)的关系,及其可能的分子机制。方法利用CCK-8、流式细胞术、Western blot和Annexin V-EGFP染色等方法,分析Res对HCT116细胞增殖和凋亡的影响。利用细胞增殖抑制实验、PCR和Western blot等实验,分析Res对HCT116细胞中BMP7表达的影响,以及BMP7对Res抑制结肠癌细胞增殖的影响及可能分子机制。结果 Res能抑制HCT116细胞增殖,诱导S期阻滞并促进其凋亡;Res增加BMP7在HCT116细胞中的mRNA和蛋白表达。过表达BMP7增强Res对HCT116细胞的增殖抑制作用,并促进凋亡,而BMP7特异性抗体则明显减弱此作用;在HCT116细胞中,Res对BMPs/Smads信号无明显影响,但能降低Akt1/2/3的磷酸化水平;过表达BMP7增强Res抑制Akt1/2/3磷酸化的作用,而BMP7特异性抗体明显减弱Res的这种作用;同时,过表达BMP7能增强Res抑制PTEN磷酸化的作用,而BMP7特异性抗体明显减弱Res对PTEN磷酸化的抑制。结论 Res对结肠癌HCT116细胞有增殖抑制作用,机制可能与Res通过上调BMP7表达而抑制PTEN磷酸化,最终使PI3K/Akt信号转导下降有关。
Aim To study the relationship between the anti-proliferative effect of resveratrol( Res) and BMP7 on human colon cancer cells and its possible molecular mechanism. Methods The proliferation of HCT116 cells was analyzed with cell proliferation inhibition assay,flow cytometry,Western blot and Annexin VEGFP staining. CCK-8,PCR and Western blot assay were used to determine the effect of Res on BMP7 in HCT116 cells and the possible molecular mechanism underlying this process. Results Res inhibited the proliferation,arrested cell cycle at S phase and promoted apoptosis in HCT116 cells. Res increased the expression of BMP7 mRNA and protein in HCT116 cells.Overexpression of BMP7 enhanced the anti-proliferative effect of Res on HCT116 cells and promoted the Resinduced apoptosis, whereas BMP7-specific antibody significantly attenuated these effects. Res exerted no apparent effect on the phosphorylation of Smad1/5/8,but decreased the phosphorylation of Akt1/2/3 substantially in HCT116 cells. Overexpression of BMP7 enhanced the inhibitory effect of Res on phosphorylation of Akt1/2/3,while BMP7 specific antibody reduced this effect notably. Res markedly decreased the phosphorylation of PTEN,which could be boosted by BMP7,but attenuated by the BMP7 specific antibody.Conclusions Res can inhibit the proliferation and promote apoptosis of HCT116 cells,and the anti-cancer activity of Res may be mediated by inactivating PI3 K/Akt signaling through up-regulating BMP7 to decrease the phosphorylation of PTEN partly.
Aim To study the relationship between the anti-proliferative effect of resveratrol( Res) and BMP7 on human colon cancer cells and its possible molecular mechanism. Methods The proliferation of HCT116 cells was analyzed with cell proliferation inhibition assay,flow cytometry,Western blot and Annexin VEGFP staining. CCK-8,PCR and Western blot assay were used to determine the effect of Res on BMP7 in HCT116 cells and the possible molecular mechanism underlying this process. Results Res inhibited the proliferation,arrested cell cycle at S phase and promoted apoptosis in HCT116 cells. Res increased the expression of BMP7 mRNA and protein in HCT116 cells.Overexpression of BMP7 enhanced the anti-proliferative effect of Res on HCT116 cells and promoted the Resinduced apoptosis, whereas BMP7-specific antibody significantly attenuated these effects. Res exerted no apparent effect on the phosphorylation of Smad1/5/8,but decreased the phosphorylation of Akt1/2/3 substantially in HCT116 cells. Overexpression of BMP7 enhanced the inhibitory effect of Res on phosphorylation of Akt1/2/3,while BMP7 specific antibody reduced this effect notably. Res markedly decreased the phosphorylation of PTEN,which could be boosted by BMP7,but attenuated by the BMP7 specific antibody.Conclusions Res can inhibit the proliferation and promote apoptosis of HCT116 cells,and the anti-cancer activity of Res may be mediated by inactivating PI3 K/Akt signaling through up-regulating BMP7 to decrease the phosphorylation of PTEN partly.
引文
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[14]Wang Z,Guo J.Mechanical induction of BMP-7 in osteocyte blocks glucocorticoid-induced apoptosis through PI3K/AKT/GSK3beta pathway[J].Cell Biochem Biophys,2013,67(2):567-74.
[15]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.
[2]Tran N H,Cavalcante L L,Lubner S J,et al.Precision medicine in colorectal cancer:the molecular profile alters treatment strategies[J].Ther Adv Med Oncol,2015,7(5):252-62.
[3]Noh J K,Naeem M,Cao J,et al.Herceptin-functionalized pure paclitaxel nanocrystals for enhanced delivery to HER2-postive breast cancer cells[J].Int J Pharm,2016,513(1-2):543-53.
[4]Liu Y Z,Wu K,Huang J,et al.The PTEN/PI3K/Akt and Wnt/beta-catenin signaling pathways are involved in the inhibitory effect of resveratrol on human colon cancer cell proliferation[J].Int J Oncol,2014,45(1):104-12.
[5]陈前昭,曾于桦,邵英,等.白藜芦醇抑制人结肠癌细胞增殖与p38 MAPK的关系研究[J].中国药理学通报,2016,32(8):1110-4.[5]Chen Q Z,Zeng Y H,Shao Y,et al.Study on the relationship between p38 MAPK and the anti-proliferation effect of resveratrol in human colon cancer cells[J].Chin Pharmacol Bull,2016,32(8):1110-4.
[6]Bragdon B,Moseychuk O,Saldanha S,et al.Bone morphogenetic proteins:a critical review[J].Cell Signal,2011,23(4):609-20.
[7]Herrera B,García-lvaro M,Cruz S,et al.BMP9 is a proliferative and survival factor for human hepatocellular carcinoma cells[J].PLo S One,2013,8(7):e69535.
[8]Peng J,Yoshioka Y,Mandai M,et al.The BMP signaling pathway leads to enhanced proliferation in serous ovarian cancer—a potential therapeutic target[J].Mol Carcinog,2016,55(4):335-45.
[9]Ren C M,Li Y,Chen Q Z,et al.Oridonin inhibits the proliferation of human colon cancer cells by upregulating BMP7 to activate p38 MAPK[J].Oncol Rep,2016,35(5):2691-8.
[10]Mundi P S,Sachdev J,Mc Court C,et al.AKT in cancer:new molecular insights and advances in drug development[J].Br J Clin Pharmacol,2016,82(4):943-56.
[11]Sadi G,Bozan D,Yildiz H B.Redox regulation of antioxidant enzymes:post-translational modulation of catalase and glutathione peroxidase activity by resveratrol in diabetic rat liver[J].Mol Cell Biochem,2014,393(1-2):111-22.
[12]Yuan S X,Wang D X,Wu Q X,et al.BMP9/p38 MAPK is essential for the antiproliferative effect of resveratrol on human colon cancer[J].Oncol Rep,2016,35(2):939-47.
[13]Sánchez-Duffhues G,Hiepen C,Knaus P,et al.Bone morphogenetic protein signaling in bone homeostasis[J].Bone,2015,80(11):43-59.
[14]Wang Z,Guo J.Mechanical induction of BMP-7 in osteocyte blocks glucocorticoid-induced apoptosis through PI3K/AKT/GSK3beta pathway[J].Cell Biochem Biophys,2013,67(2):567-74.
[15]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.