摘要
目的:研究养阴解毒方对人肺腺癌耐药细胞PC9/R信号蛋白AKT磷酸化及抑癌基因PTEN的表达影响。方法:选择吉非替尼获得性耐药细胞株PC9/R,采用血清药理学的方法,研究养阴解毒方及其结合吉非替尼的体外增殖抑制作用,流式细胞术法检测对PC9/R的细胞凋亡及周期的影响;Western-blot法检测PC9/R细胞株的PI3K/Akt信号通路中的关键激酶Akt磷酸化和PTEN表达。结果:养阴解毒方对耐药株PC9/R有增殖抑制作用,并有量效关系,低剂量组联合吉非替尼抑制PC9/R效果最优(P<0.01),增效作用明显(Q值均>0.85);养阴解毒联和吉非替尼组较吉非替尼单药组可明显提高PC9/R细胞凋亡的能力,有显著统计学意义(P<0.01),养阴解毒方组及联合用药组对PC9/R细胞S期均较吉非替尼单药细胞周期缩短(P<0.01);养阴解毒方组与联合用药组均比单药吉非替尼能够明显抑制PC9/R的p-AKT活化位点Thr308和Ser473的磷酸化,有统计学意义(P均<0.05),联合用药组PC9/R的PTEN表达显著高于其余各组,有统计学意义(P均<0.05)。结论:养阴解毒方可增加人肺腺癌耐药株细胞PC9/R对于吉非替尼的药物敏感性,两药联用有协同作用,养阴解毒方可提高PTEN的表达,抑制信号通路关键蛋白AKT的磷酸化,从而起到抑制肿瘤生长、逆转耐药的双重作用。
Objective:To study the effect of Yangyin Jiedu Formula on phosphorylation of signaling protein AKT and tumor suppressor gene PTEN in human lung adenocarcinoma cells PC9/R.Methods:Cell line PC9/R of Gefitinib-acquired resista nee was selected,serum pharmacology was used to study the inhibitory effect on PC9/R of Yangyin Jiedu Formula and its combination with Gefitinib in vitro.Flow cytometry was used to detect PC9/R cell apoptosis and the cell cycles influenced by the formula.The phosphorylation of key kinase Akt and PTEN expression in the PI3K/Akt signaling pathway of PC9/R cell line were detected by Western-blot assay.Results:Yangyin Jiedu Formula had a proliferation-inhibiting effect on drug-resistant strain PC9/R,which showed dose-effect relationship,the low-dose group combined with Gefitinib inhibited PC9/R with the best effect(P<0.01),and the synergistic effect was obvious(Q>0.85).Yangyin Jiedu Formula combined with Gefitinib can significantly promoted apoptosis of the PC9/R cell compared to Gefitinib monotherapy(P<0.01).Single cell cycle in PC9/R cells was shortened in Yangyin Jiedu Formula group and in the combination group,when compared to that in the Gefitinib group(P<0.01).The phosphorylation of p-AKT activation sites Thr308 and Ser473 in PC9/R significantly inhibited in Yangyin Jiedu Formula group and in the combination group compared to that in Gefitinib group(P<0.05).The PTEN expression of PC9/R in the combination group was significantly higher than that in the other two groups(P<0.05).Conclusion:Yangyin Jiedu Formula can increase the sensitivity of human lung adenocarcinoma cell line PC9/R to Gefitinib.The combination of the two dmgs has a synergistic effect.Yangyin Jiedu Formula can increase the expression of PTEN and inhibit the Phosphorylation of the key signal protein AKT.It plays a dual role in inhibiting tumor growth and reversing drug resistance.
作者
张婷
李昂
孙建立
ZHANG Ting;LI Ang;SUN Jianli(Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200030,China;Huashan Hospital Affiliated to Fudan University,Shanghai 264000,China;Wuhan Hannan District Peoples Hospital,Wuhan 430090,China)
出处
《中医药学报》
CAS
2020年第1期11-16,共6页
Acta Chinese Medicine and Pharmacology
基金
国家自然科学基金面上项目(81573890)
国家中医药管理局中医临床研究基地业务建设科研专题课题(JDZX2015069)
刘嘉湘国医大师传承工作室建设项目(DSGZS-2017002)
龙华医院中青年名中医培养项目。
作者简介
张婷(1987-),女,硕士,主治医师,主要从事中医药防治恶性肿瘤的临床研究;通讯作者:孙建立(1966-),男,博士,主任医师,硕士研究生导师,主要从事中西医结合治疗肺恶性肿瘤的临床和实验研究。
