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目的初步探讨阻断程序性死亡-配体1(programmed death-ligand 1,PD-L1)在增强奥沙利铂(oxaliplatin,L-OHP)治疗原发性肝细胞肝癌(hepatocellular carcinoma,HCC)疗效中的作用及其分子机制。方法 L-OHP处理人HCC细胞系HepG2后,q RT-PCR和Western blot检测PD-L1在m RNA及蛋白水平的表达变化,以及细胞外调节蛋白激酶1/2(extracellular regulated protein kinases,ERK1/2)磷酸化水平的改变;建立Hepa1-6细胞C57BL/6J小鼠HCC移植瘤模型,随机分为对照组、L-OHP组、PD-L1单抗组、L-OHP+PD-L1单抗组,计算处理后各组小鼠肿瘤的质量、抑制率和生存时间;体外刺激或抑制HepG2细胞系ERK1/2的磷酸化,L-OHP处理后q RT-PCR和Western blot检测PD-L1的表达变化。结果 L-OHP能显著抑制HepG2细胞系的生长,其IC50分别为20.94 mg/L;L-OHP处理HepG2细胞后,p-ERK1/2和PD-L1的表达量均高于对照组(P<0.05)。小鼠HCC移植瘤模型中,与对照组、L-OHP组和PD-L1单抗组相比较,L-OHP+PD-L1单抗组的肿瘤质量显著减小(P<0.05),肿瘤抑制率大幅提升(P<0.05),生存时间明显延长(P<0.05)。体外刺激ERK1/2的磷酸化后,PD-L1表达显著上调,而抑制HCC细胞株ERK1/2的磷酸化后,L-OHP诱导PD-L1表达升高效应明显受到抑制。结论 L-OHP能通过提高ERK1/2磷酸化水平上调HCC细胞的PD-L1表达,从而导致治疗失败,而联合PD-L1单抗可显著增强L-OHP的抗HCC作用。
Objective To investigate the effect of blocking programmed death-ligand 1 (PD-L1) on the treatment of primary hepatocellular carcinoma (HCC) with oxaliplatin (L-OHP) In the role and molecular mechanism. Methods The expression of PD-L1 in m RNA and protein level and the expression of extracellular regulated protein kinase (ERK1 / 2) in human HCC cell line HepG2 were detected by q RT-PCR and Western blot. 2) phosphorylated Hpa xenografts in C57BL / 6J mice were randomly divided into control group, L-OHP group, PD-L1 monoclonal antibody group and L-OHP + PD-L1 monoclonal antibody Group, the tumor mass, inhibition rate and survival time of mice in each group were calculated; the phosphorylation of ERK1 / 2 in HepG2 cell line was stimulated or inhibited in vitro; the expression of PD-L1 was detected by q RT-PCR and Western blot after L-OHP treatment Change of expression Results L-OHP significantly inhibited the growth of HepG2 cells with IC50 of 20.94 mg / L, respectively. The expression of p-ERK1 / 2 and PD-L1 in HepG2 cells treated with L-OHP was higher than that in control group (P < 0.05). Compared with the control group, L-OHP group and PD-L1 monoclonal antibody group, the tumor mass of L-OHP + PD-L1 monoclonal antibody group was significantly decreased (P <0.05), tumor suppression The rate of survival increased significantly (P <0.05) and the survival time was significantly prolonged (P <0.05). The phosphorylation of ERK1 / 2 stimulated the phosphorylation of ERK1 / 2 in vitro, and the expression of PD-L1 was significantly up-regulated. The phosphorylation of ERK1 / 2 inhibited the phosphorylation of ERK1 / 2 in HCC cells. Conclusion L-OHP can up-regulate the expression of PD-L1 in HCC cells by increasing the phosphorylation of ERK1 / 2, which leads to failure of treatment. Combined with PD-L1 monoclonal antibody can significantly enhance the anti-HCC effect of L-OHP.