目的 探讨ATP诱导免疫原性细胞死亡在肝癌免疫应答中的作用及其机制.方法 人肝癌细胞系HepG2购自于上海弘顺生物科技有限公司,经培养及传代后,随机分为对照组和ATP组.ATP组中采用6 mmol/L ATP的培养液孵育HepG2细胞24 h;对照组将处于对数期HepG2细胞常规培养.CCK8实验检测HepG2细胞毒性实验;流式细胞仪检测HepG2细胞的凋亡;免疫荧光技术检测HepG2细胞的钙网蛋白膜转位;酶联免疫吸附试验(ELISA)检测ATP和高迁移率族蛋白1(HMGB1)含量;蛋白质印迹法检测β2微球蛋白(β2m)、TAP相关糖蛋白(TAPBP)、抗原处理相关转运体(Tap)1、Tap2和鸟苷酸腺苷酸合成酶-干扰素基因刺激因子(cGAS-STING)通路蛋白表达;Real-timePCR检测PD-L1和ICAM-1 mRNA的表达.结果 6 mmol/L的ATP的作用下,能使HepG2细胞的抑制率达到50％;ATP组人肝癌HepG2细胞凋亡率显著高于对照组,差异有统计学意义(P<0.05).与对照组相比,ATP组中钙网蛋白明显发生膜转位;且ATP组中HepG2细胞释放ATP和HMGB1的表达量明显高于对照组,差异均有统计学意义(P<0.05).ATP组中β2m、Tapbp、Tap1、Tap2蛋白表达明显高于对照组,差异均有统计学意义(P<0.05).ATP组中PD-L1 mRNA表达显著低于对照组,ICAM-1 mRNA表达显著高于对照组,差异均有统计学意义(P<0.05).ATP组中cGAS和STING蛋白表达明显高于对照组,差异均有统计学意义(P<0.05).结论 ATP可以通过激活cGAS-STING信号通路,诱导肝癌细胞免疫原性细胞死亡的发生,增强肝癌的免疫原性,激活抗肿瘤免疫.

目的 分析miR-218-5p在非小细胞肺癌中的表达及其对TPD52的靶向调控作用.方法 回顾性收集2020年9月至2021年12月青岛大学附属青岛市中心医院行手术切除的治疗NSCLC患者39例,术中收取患者的癌组织及癌旁组织,实时定量PCR检测miR-218-5p表达,体外培养H1975细胞株,将H1975细胞分为NC组(mimics NC转染)、miR-218-5p组(miR-218-5p mimics转染)、TPD52-siRNA组(TPD52-siRNA转染)和miR-218-5p+TPD52-siRNA组(转染miR-218-5p mimics后再加入TPD52-siRNA试剂进行转染).平板克隆形成实验检测H1975细胞的克隆形成数目,Transwell检测H1975细胞侵袭,划痕实验检测H1975细胞迁移,双荧光素酶实验测定miR-218-5p和TPD52靶向关系,蛋白质印迹检测TPD52蛋白表达.结果 NSCLC组织中miR-218-5p的表达明显低于癌旁组织,差异有统计学意义(P<0.05).miR-218-5p的表达与性别、年龄、吸烟无相关性(P>0.05),但与淋巴结是否转移、肿瘤大小、血管是否侵犯、TNM分期有关(P<0.05).miR-218-5p组克隆形成数目、侵袭率和迁移率显著低于NC组,差异均有统计学意义(P<0.05).与NC组相比,miR-218-5p显著抑制了TPD523'UTR-WT报告基因的荧光表达,差异有统计学意义(P<0.05).miR-218-5p组中TPD52蛋白的表达明显低于NC组,差异均有统计学意义(P<0.05).miR-218-5p组和TPD52-siRNA组的克隆形成数目、侵袭率和迁移率显著低于NC组,差异均有统计学意义(P<0.05);miR-218-5p+TPD52-siRNA组的克隆形成数目、迁移率和侵袭率显著低于miR-218-5p组和TPD52-siRNA组,差异均有统计学意义(P<0.05).结论 miR-218-5p在NSCLC组织中高表达,能够抑制NSCLCH1975细胞克隆形成、侵袭和迁移能力,并且通过靶向TPD52发挥着抑癌作用.

Objective:To observe the efficacy and safety of camrelizumab combined with albumin-paclitaxel/lobaplatin as neoadjuvant therapy for locally advanced esophageal cancer.Methods:From 2019 to 2020, 18 patients with locally advanced esophageal squamous cell carcinoma who were initially unresectable were treated with neoadjuvant therapy of camrelizumab combined with albumin-paclitaxel/lobaplatin for 3-4 cycles before operation, and the operation was evaluated after treatment. The surgical removal rate (R 0), pathological complete response (pCR), major pathological response (MPR), the neoadjuvant objective response rate (ORR) and safety were observed. Results:The median age of 18 patients was 59 years (32-66 years). The neoadjuvant treatment completion rate was 89% (16/18). The R 0 rate was 61% (11/18), with pCR 45% (5/11), MPR 64% (7/11) and ORR 69%. The incidence of adverse reactions were neutropenia (78%), thrombocytopenia (61%) and reactive cutaneous capillary endothelial proliferation (61%), but the main adverse reactions were grade 1-2. The postoperative complications were mild. Conclusion:As a neoadjuvant therapy, camrelizumab combined with albumin-paclitaxel/lobaplatin can improve the pathological response rate of locally advanced esophageal cancer with good safety.