溶瘤腺病毒rAd.DCN对非小细胞肺癌细胞生长的抑制作用  

Inhibitory effect of oncolytic adenovirus rAd.DCN on the growth of non-small cell lung cancer cells

作　　者：赵慧强 易静 杜海涛 韩巧君 王建斌 欧敏 朱艳玲 ZHAO Huiqiang;YI Jing;DU Haitao;HAN Qiaojun;WANG Jianbin;OU Min;ZHU Yanling(National Clinical Research Center for Geriatric Diseases,Healthcare Department Six,the Second Medical Center,Chinese PLA General Hospital,Beijing 100048,China;Institute of Radiation Medicine,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100850,China;Department of Ultrasound Diagnostics,No.5 Campus of Qingdao Special Servicemen Recuperation Center of PLA Navy,Qingdao,Shandong 266071,China)

机构地区：[1]解放军总医院第二医学中心保健六科,国家老年疾病临床医学研究中心,北京100048 [2]军事科学院军事医学研究院辐射医学研究所,北京100850 [3]海军青岛特勤疗养中心疗养五区超声诊断科,山东青岛266071

出　　处：《军事医学》2023年第7期508-514,共7页Military Medical Sciences

基　　金：解放军总医院第六医学中心创新培育基金(CXPY201904)。

摘　　要：目的评价rAd.DCN对NCI-H292、NCI-H1703和NCI-H1975三种不同非小细胞肺癌(NSCLC)细胞生长的抑制作用。方法将3种NSCLC细胞培养扩增至对数生长期,每种细胞分为3组:空白对照组、rAd.Null处理组和rAd.DCN处理组。感染后48 h利用Western印迹方法检测核心蛋白聚糖(Decorin)的表达情况;感染后72 h使用SRB法检测NSCLC细胞感染病毒后的存活率,评价其杀伤效率;感染后24、48、72和96 h使用CCK8法测定各组经不同处理后NSCLC细胞的活性,评价其增殖情况;感染后48 h使用实时荧光定量PCR(qPCR)检测Decorin靶基因的表达情况。结果rAd.DCN可有效感染3种NSCLC细胞,高表达Decorin蛋白。rAd.Null和rAd.DCN感染NSCLC细胞后均可直接杀伤肿瘤细胞,降低肿瘤细胞存活率,且杀伤力与病毒感染复数相关;rAd.DCN的效果优于rAd.Null,二者对3种NSCLC细胞杀伤率之间的差异具有统计学意义(P<0.05)。与对照组相比,rAd.Null和rAd.DCN可明显抑制NSCLC细胞增殖;与rAd.Null处理组相比,rAd.DCN处理组细胞增殖抑制作用更强,二者间差异显著(P<0.0001)。rAd.DCN感染NSCLC后,其靶基因CTNNB-1、表皮生长因子受体(EGFR)、间质表皮转化因子(MET)和转化生长因子β(TGF-β)表达均下调。结论rAd.DCN感染NSCLC细胞后,可通过直接杀伤、抑制增殖和下调靶基因表达等不同途径抑制NSCLC细胞生长,有望成为新的靶向治疗药物。Objective To evaluate the inhibitory effect of rAd.DCN on the growth of NCI-H292,NCI-H1703 and NCI-H1975 non-small cell lung cancer(NSCLC)cells.Methods Three types of NSCLC cells were expanded to the exponential growth stage before each type was divided into three groups:blank control group,rAd.Null treatment group and rAd.DCN treatment group.The expression of Decorin was detected by Western blotting 48 hours after infection.The survival rate of NSCLC cells infected with virus was detected by SRB method 72 hours after infection,and the killing efficiency was evaluated.At 24,48,72 and 96 hours post infection,CCK8 assay was used to measure the activity of NSCLC cells after different treatments,and the proliferation of NSCLC cells was evaluated.The expressions of target genes of Decorin were detected by quantitative real-time polymerase chain reaction(qPCR)48 hours after infection.Results rAd.DCN could effectively infect three types of NSCLC cells and highly express Decorin protein.rAd.Null and rAd.DCN infection directly killed NSCLC tumor cells and reduced the survival rate of tumor cells,and the killing ability was related to multiplicity of infection.The killing effect of rAd.DCN was better than that of rAd.Null,and the difference in killing rates between rAd.Null and rAd.DCN on three NSCLC cells was statistically significant(P<0.05).Compared to the control group,rAd.Null and rAd.DCN could inhibit the proliferation of NSCLC cells.Compared with the rAd.Null treatment group,the rAd.DCN treatment group had a stronger inhibitory effect on cell proliferation,and there was significant difference between the two groups(P<0.0001).The expressions of target genes of rAd.DCN,catenin beta 1(CTNNB-1),epidermal growth factor receptor(EGFR),mesenchymal-epithelial transition factor(MET)and transforming growth factor-β(TGF-β)were down-regulated in NSCLC after being infected with rAd.DCN.Conclusion rAd.DCN can inhibit the growth of NSCLC cells via direct killing,inhibition of proliferation and down-regulation of target gene expr

关 键 词：核心蛋白聚糖 非小细胞肺癌细胞 生长 

分 类 号：R734.2[医药卫生—肿瘤]