热疗通过靶向HOXB1诱导非小细胞肺癌细胞凋亡的研究  

Study on themotherapyin inducing cell apoptosis of non-small cell lung cancer cells by targeting HOXB1

作　　者：刘威[1] 张帆[1] 龙永贵[1] LIU Wei;ZHANG Fan;LONG Yonggui(Department of Thoracic Surgery,Leshan Municipal People′s Hospital,Leshan,Sichuan 614000,China)

机构地区：[1]四川省乐山市人民医院胸外科,四川乐山614000

出　　处：《国际检验医学杂志》2022年第24期3037-3042,共6页International Journal of Laboratory Medicine

摘　　要：目的探讨热疗调控非小细胞肺癌(NSCLC)细胞凋亡的潜在机制。方法在37、40、43、46℃的温度下处理NSCLC细胞系A549、HCC827、NCI-H1299,并分析热疗对NSCLC细胞活力和凋亡水平的影响。将37、43℃处理的NSCLC细胞系进行转录组高通量测序并分析3种细胞系中同时被调控的基因。通过遗传学筛选鉴定热疗调控NSCLC细胞凋亡的关键分子同源盒结构基因B1(HOXB1)。siGFP或siHOXB1敲低后检测NSCLC细胞系在43℃下的凋亡水平。过表达Vector或HOXB1后检测NSCLC细胞系在37℃下的凋亡水平。siGFP或siHOXB1敲低后采用荧光素酶报告实验检测NF-κB的活性。结果相比于37℃,NSCLC细胞系在40、43、46℃下的活力均显著降低(P<0.05),凋亡水平均显著上升(P<0.05)。相比于37、40、46℃下,NSCLC细胞系在43℃的凋亡水平显著上升(P<0.05)。与siGFP组比较,敲低HOXB1后,43℃时NSCLC细胞系A549、HCC827、NCI-H1299的凋亡水平下降(P<0.05)。与Vector组比较,过表达HOXB1后,37℃时NSCLC细胞系A549、HCC827、NCI-H1299的凋亡水平上升(P<0.05)。与siGFP组比较,敲低HOXB1后,43℃时NSCLC细胞系中荧光素酶活性上升(P<0.05)。43℃条件下,免疫共沉淀通过HOXB1抗体能够结合NSCLC细胞系A549、HCC827、NCI-H1299的NF-κB。结论热疗处理NSCLC细胞后,HOXB1表达水平上升,HOXB1可与NF-κB结合并抑制NF-κB的活性,促进了NSCLC细胞凋亡。Objective To explore the potential mechanism of themotherapy in regulating cell apoptosis in non-small cell lung cancer(NSCLC).Methods NSCLC cell lines A549,HCC827 and NCI-H1299 were treated at 37,40,43 and 46℃,and the effects of themotherapy on the viability and apoptosis levels of NSCLC cells were analyzed.The NSCLC cell lines treated at 37 and 43℃conducted the transcriptome high-throughput sequencing and the genes that were simultaneously regulated in the three kinds of cells were analyzed.The key molecular homology cassette structural gene B1(HOXB1)regulating apoptosis in NSCLC cells by themotherapy was identified by genetic screening.After siGFP or siHOXB1 knockdown,the apoptosis level of NSCLC cell lines at 43℃was detected.The apoptosis levels of NSCLC cell lines at 37℃were measured after overexpression of Vector or HOXB1.After siGFP or siHOXB1 knockdown,the luciferase reporter assay was used to detect NF-κB activity.Results The viability of NSCLC cell lines at 40,43 and 46℃were significantly decreased compared with at 37℃(P<0.05)and the apoptosis levels were significantly increased(P<0.05).Compared with at 37,40 and 46℃,the apoptosis level of NSCLC cell lines at 43℃was significantly increased(P<0.05).Compared with the siGFP group,after kockdown of HOXB1,the apoptosis of A549,HCC827 and NCI-H1299 in NSCLC cell line at 43℃was decreased(P<0.05).Compared with the Vector group,the apoptosis levels of A549,HCC827 and NCI-H1299 in NSCLC cell lines at 37℃were increased after overexpression of HOXB1(P<0.05).Compared with the siGFP group,after knockdown of HOXB1,the luciferase activity in NSCLC cell lines at 43℃was increased(P<0.05).The immunoprecipitation by HOXB1 antibody was able to bind NF-κB of A549,HCC827 and NCI-H1299 in NSCLC cell lines.Conclusion After treating NSCLC cells by themotherapy,the expression level of HOXB1 is increased.HOXB1 can bind to NF-κB and inhibit the activity of NF-κB,and promote NSCLC cell apoptosis.

关 键 词：热疗 HOXB1 非小细胞肺癌 凋亡 核因子-ΚB 

分 类 号：R734.2[医药卫生—肿瘤]