迷迭香酸衍生物RAD-9通过PI3K/Akt和p38 MAPK信号通路诱导胃癌MGC-803细胞凋亡
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摘要
目的研究迷迭香酸衍生物RAD-9诱导胃癌MGC-803细胞凋亡的作用及其机制。方法 MTT法观察RAD-9对胃癌MGC-803细胞增殖的抑制作用;流式细胞术检测细胞的凋亡;Hoechst 33258染色法观察RAD-9对MGC-803细胞核凋亡形态学的影响;Western blot检测RAD-9干预MGC-803细胞36 h后,对Akt、p-Akt、p38 MAPK、p-p38 MAPK蛋白及凋亡相关蛋白Bcl-2、Bax、caspase-3的影响。结果MTT结果显示,RAD-9呈时间、浓度依赖性抑制胃癌MGC-803细胞增殖;流式细胞术结果显示,RAD-9对胃癌MGC-803细胞有明显的促凋亡作用(P<0.01);Hoechst 33258染色实验结果显示,RAD-9干预胃癌MGC-803细胞36 h后,细胞核呈现典型凋亡形态学改变;Western blot结果显示,RAD-9干预胃癌MGC-803细胞36 h后,Bcl-2蛋白表达水平明显降低,Bax、caspase-3蛋白表达水平明显提高,Akt、p-Akt蛋白表达水平明显下调,p38 MAPK、p-p38 MAPK蛋白表达水平明显上调(P<0.01)。结论 RAD-9能抑制胃癌MGC-803细胞生长,且能诱导其凋亡,其机制可能与抑制PI3K/Akt和激活p38 MAPK信号通路相关。
Aim To study the apoptosis-inducing effect of rosmarinic acid derivative RAD-9 on gastric cancer MGC-803 cells and the underlying mechanisms. Methods MTT assay was taken to detect the survival of gastric cancer MGC-803 cells effected by RAD-9. Cell apoptosis was detected by flow cytometry. The apoptotic morphology of MGC-803 cells was observed by Hoechst 33258 staining. The protein expression levels of Bcl-2,Bax,caspase-3,Akt,p-Akt,p38 MAPK and p-p38 MAPK were measured by Western blot. Results The results of MTT assay showed that RAD-9 inhibited the viability of gastric cancer MGC-803 cells in a time and concentration-dependent manner. Flow cytometry showed that RAD-9 significantly promoted apoptotic cell percentage in gastric cancer MGC-803 cells( P < 0. 01). Hoechst 33258 staining showed that the nucleus of MGC-803 cells could be observed with typical apoptotic morphological changes after RAD-9 administration. Compared with the control group, the protein expression levels of Bcl-2,Akt,p-Akt were significantly down-regulated( P < 0. 01),while those of Bax,caspase-3,p38 MAPK,p-p38 MAPK were significantly up-regulated( P < 0. 01). Conclusion RAD-9 can inhibit the growth and further induce apoptosis in gastric cancer MGC-803 cells,which may involve inhibiting PI3 K/Akt and activating p38 MAPK signaling pathway.
Aim To study the apoptosis-inducing effect of rosmarinic acid derivative RAD-9 on gastric cancer MGC-803 cells and the underlying mechanisms. Methods MTT assay was taken to detect the survival of gastric cancer MGC-803 cells effected by RAD-9. Cell apoptosis was detected by flow cytometry. The apoptotic morphology of MGC-803 cells was observed by Hoechst 33258 staining. The protein expression levels of Bcl-2,Bax,caspase-3,Akt,p-Akt,p38 MAPK and p-p38 MAPK were measured by Western blot. Results The results of MTT assay showed that RAD-9 inhibited the viability of gastric cancer MGC-803 cells in a time and concentration-dependent manner. Flow cytometry showed that RAD-9 significantly promoted apoptotic cell percentage in gastric cancer MGC-803 cells( P < 0. 01). Hoechst 33258 staining showed that the nucleus of MGC-803 cells could be observed with typical apoptotic morphological changes after RAD-9 administration. Compared with the control group, the protein expression levels of Bcl-2,Akt,p-Akt were significantly down-regulated( P < 0. 01),while those of Bax,caspase-3,p38 MAPK,p-p38 MAPK were significantly up-regulated( P < 0. 01). Conclusion RAD-9 can inhibit the growth and further induce apoptosis in gastric cancer MGC-803 cells,which may involve inhibiting PI3 K/Akt and activating p38 MAPK signaling pathway.
引文
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[2]Yuan L W,Yamashita H,Seto Y.Glucose metabolism in gastric cancer:the cutting-edge[J].World J Gastroenterol,2016,22(6):2046-59.
[3]Hu Q,Sun W,Wang C,et al.Recent advances of cocktail chemotherapy by combination drug delivery systems[J].Adv Drug Deliv Rev,2016,98:19.
[4]牟宜双,邓文龙,周黎明.迷迭香酸抗肿瘤作用研究进展[J].中药药理与临床,2015,31(1):266-9.[4]Mou Y S,Deng W L,Zhou L M.Research progress of antitumor effect of rosmarinic acid[J].Pharmacol Clin Chin Mater Med,2015,31(1):266-9.
[5]郭峰,朱炳阳,迟秀玲,等.迷迭香酸抗过氧化氢诱导血管平滑肌细胞凋亡作用的研究[J].中国药理学通报,2007,23(3):365-70.[5]Guo F,Zhu B Y,Chi X L,et al.Inhibition of rosmarinic acid on the apoptosis of vascular smooth muscle cells induced by hydrogen peroxide[J].Chin Pharmacol Bull,2007,23(3):365-70.
[6]Zhou L,Zhang P,Zhai C,et al.Anti-Warburg effect of rosmarinic acid via miR-155 in gastric cancer cells[J].Drug Des Devel T-her,2015,9:2695-703.
[7]Chen W,Zheng R,Baade P D,et al.Cancer statistics in China,2015[J].CA Cancer J Clin,2016,66(2):115.
[8]陈凛,张珂诚.转移性胃癌的外科治疗[J].中华胃肠外科杂志,2017,20(7):731-4.[8]Chen L,Zhang K C.Surgical treatment for metastatic gastric cancer[J].Chin J Gastrointest Surg,2017,20(7):731-4.
[9]Gao L,Hao J,Niu Y Y,et al.Network pharmacology dissection of multiscale mechanisms of herbal medicines in stage IV gastric adenocarcinoma treatment[J].Medicine,2016,95(35):e4389.
[10]Peng Y T,Chen P,Ouyang R Y,et al.Multifaceted role of prohibitin in cell survival and apoptosis[J].Apoptosis,2015,20(9):1135-49.
[11]Li B,Li J,Xu W W,et al.Suppression of esophageal tumor growth and chemoresistance by directly targeting the PI3K/AKTpathway[J].Oncotarget,2014,5(22):11576-87.
[12]Siddiqui W A,Ahad A,Ahsan H.The mystery of BCL2 family:Bcl-2 proteins and apoptosis:an update[J].Arch Toxicol,2015,89(3):289-317.
[13]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.[13]Chen Q Z,Zeng Y Y,Shao Y,et al.Anti-proliferation effect of resveratrol and p38 MAPK in human colon cancer cells[J].Chin Pharmacol Bull,2016,32(8):1110-4.
[14]Ghatan S,Larner S,Kinoshita Y,et al.p38 MAP kinase mediates Bax translocation in nitric oxide-induced apoptosis in neurons[J].J Cell Biol,2000,150(2):335-47.
[15]梁先敏,杨克敌.Caspase和JNK/SAPK、p38 MAPK与细胞凋亡[J].国外医学卫生学分册,2008,35(1):5-10.[15]Liang X M,Yang K D.Caspase and JNK/SAPK,p38 MAPK and apoptosis[J].Foreign Med Sci(Sect Hyg),2008,35(1):5-10.
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