地西他滨联合丙戊酸钠促进肝癌细胞凋亡的ERK通路研究
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摘要
目的:本研究对地西他滨联合丙戊酸钠促进肝癌细胞凋亡的相关机制进行研究,为临床治疗和新药研发提供参考。方法:肝癌细胞HepG2经地西他滨联合丙戊酸钠孵育后,MTT法检测二者对HepG2细胞的抑制率,Real-time PCR及免疫印迹法检测细胞凋亡蛋白Caspase3/9、Bax及Bcl2和ERK信号通路关键蛋白的表达情况。结果:地西他滨联合丙戊酸钠能够有效抑制肝癌细胞的增殖,且二者联用的抑制率明显高于单独应用(P<0.05),同时与对照组相比,细胞凋亡蛋白Caspase3、Caspase9及Bax表达明显增强(P<0.05),Bcl2表达明显降低(P<0.05),且ERK信号通路关键蛋白Ras、Raf及MEK和ERK1/2的表达明显升高,与对照组细胞有显著的统计学差异性(P<0.05)。结论:地西他滨联合丙戊酸钠主要通过激活MEK/ERK信号转导通路抑制肝癌细胞HepG2的增殖过程。
Objective:The present research aimed to explore the involvement of ERK signaling pathway in apoptosis of hepatic carcinoma induced by decitabine combined sodium valproate.Methods:HepG2 cell line was incubated with decitabine combined sodium valproate and the inhibition rate was detected by MTT method.The apoptotic related protein and ERK signal pathway proteins were assayed by Real-time PCR or Western blot.Results:The HepG2 cell line was inhibited greatly by decitabine combined sodium valproate medication with an increased expression of Caspase3,Caspase9 and Bax(P < 0.05).The expression of Ras,Raf,MEK and ERK1 /2 was increased dramatically after incubation with decitabine combined sodium valproate when compared with control group(P< 0.05).Conclusion:Decitabine combined sodium valproate exert a significant inhibition on the proliferation of HepG2 cell line by normalizing the abnormal MEK/ERK signaling pathway.
Objective:The present research aimed to explore the involvement of ERK signaling pathway in apoptosis of hepatic carcinoma induced by decitabine combined sodium valproate.Methods:HepG2 cell line was incubated with decitabine combined sodium valproate and the inhibition rate was detected by MTT method.The apoptotic related protein and ERK signal pathway proteins were assayed by Real-time PCR or Western blot.Results:The HepG2 cell line was inhibited greatly by decitabine combined sodium valproate medication with an increased expression of Caspase3,Caspase9 and Bax(P < 0.05).The expression of Ras,Raf,MEK and ERK1 /2 was increased dramatically after incubation with decitabine combined sodium valproate when compared with control group(P< 0.05).Conclusion:Decitabine combined sodium valproate exert a significant inhibition on the proliferation of HepG2 cell line by normalizing the abnormal MEK/ERK signaling pathway.
引文
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[13]柯晴,岑洪,胡晓桦.去甲基化药物地西他滨治疗血液系统恶性肿瘤的研究进展[J].医学综述,2010,11(7):1068-1070,1121.
[14]吴凤娟,Koirala D,刘斯婕,等.新型噻唑并三嗪类化合物R001抑制ERK1/2磷酸化的抗肿瘤作用[J].现代生物医学进展,2012,16(29):5638-5642.
[15]郝志楠,郑勇斌,肖高春,等.丝裂原活化蛋白激酶/细胞外调节蛋白激酶信号通路与肿瘤血管新生的关系[J].中国医药导报,2015,8(1):148-151.
[16]郝杰,程澎,王艳煜,等.地西他滨联合预激方案和传统治疗方案治疗老年MDS/MPD转化的急性髓系白血病的疗效和安全性的比较研究[J].临床血液学杂志,2014,5(1):15-18.
[17]方宝枝,何广胜,吴德沛,等.地西他滨治疗成人急性髓细胞白血病的临床观察[J].肿瘤,2011,14(11):1022-1025.
[2]Yang HJ,Lee JH,Lee DH,et al.Small single-nodule hepatocellular carcinoma:comparison of transarterial chemoembolization,radiofrequency ablation,and hepatic resection by using inverse probability weighting[J].Radiology,2014,271(3):909-918.
[3]Lin HW,Tseng FY.Ectopic adrenalcorticotropic hormone syndrome improved by transarterial embolization to hepatic metastatic lesions of pancreatic neuroendocrine carcinoma:a case report[J].J Clin Oncol,2012,30(33):e360-e363.
[4]朱乔,陈春伟,巩丽,等.CSMD1在肝细胞肝癌中的研究进展[J].现代肿瘤医学,2014,7(11):2760-2763.
[5]Lloyd SA,Lewis GS,Zhang Y,et al.Connexin 43 deficiency attenuates loss of trabecular bone and prevents suppression of cortical bone formation during unloading[J].J Bone Miner Res,2012,27(11):2359-2372.
[6]姚亮,李慧玲,董建一,等.低水平活性氧对Ras癌基因诱导肝肿瘤发生的影响[J].中华肿瘤防治杂志,2015,6(12):911-916.
[7]张丽志,瞿全新,温克.ERK信号转导通路对肿瘤细胞生物学行为的调控[J].医学综述,2011,9(6):836-838.
[8]Fang XX,Jiang XL,Han XH,et al.Neuroprotection of interleukin-6 against NMDA-induced neurotoxicity is mediated by JAK/STAT3,MAPK/ERK,and PI3K/AKT signaling pathways[J].Cell Mol Neurobiol,2013,33(2):241-251.
[9]Ferraro A,Mourtzoukou D,Kosmidou V,et al.EZH2 is regulated by ERK/AKT and targets integrin alpha2 gene to control Epithelial-Mesenchymal Transition and anoikis in colon cancer cells[J].Int J Biochem Cell Biol,2013,45(2):243-254.
[10]杨浩,杨连粤.肝癌术后复发转移相关因素研究进展[J].中国实用外科杂志,2012,22(10):868-870,876.
[11]王帅兵,杜宝昌.双膦酸盐在恶性肿瘤治疗中的作用研究进展[J].肿瘤学杂志,2015,6(1):66-69.
[12]张丙银,张传东,刘艳军,等.济宁市市中区2012年恶性肿瘤发病和死亡分析[J].中华肿瘤防治杂志,2015,7(8):574-578.
[13]柯晴,岑洪,胡晓桦.去甲基化药物地西他滨治疗血液系统恶性肿瘤的研究进展[J].医学综述,2010,11(7):1068-1070,1121.
[14]吴凤娟,Koirala D,刘斯婕,等.新型噻唑并三嗪类化合物R001抑制ERK1/2磷酸化的抗肿瘤作用[J].现代生物医学进展,2012,16(29):5638-5642.
[15]郝志楠,郑勇斌,肖高春,等.丝裂原活化蛋白激酶/细胞外调节蛋白激酶信号通路与肿瘤血管新生的关系[J].中国医药导报,2015,8(1):148-151.
[16]郝杰,程澎,王艳煜,等.地西他滨联合预激方案和传统治疗方案治疗老年MDS/MPD转化的急性髓系白血病的疗效和安全性的比较研究[J].临床血液学杂志,2014,5(1):15-18.
[17]方宝枝,何广胜,吴德沛,等.地西他滨治疗成人急性髓细胞白血病的临床观察[J].肿瘤,2011,14(11):1022-1025.