PD-L1基因沉默对多柔比星诱导乳腺癌SK-BR3细胞自噬的影响  

The effect of PD-L1 gene silencing on Doxorubicin-induced autophagy in breast cancer SK-BR3 cells

作　　者：林小洪 Lin Xiaohong(Department of Clinical laboratory,Pingtan Comprehensive Experimental Zone Hospital,Fuzhou,Fujian 350400,China)

机构地区：[1]平潭综合实验区中医院检验科,福建福州350400

出　　处：《齐齐哈尔医学院学报》2025年第8期730-735,共6页Journal of Qiqihar Medical University

摘　　要：目的探讨程序性死亡配体1(PD-L1)基因沉默对多柔比星(DOX)诱导乳腺癌SK-BR3细胞自噬的影响。方法通过Western blot(WB)和实时荧光定量PCR检测技术证实SK-BR3细胞表达PD-L1,利用CRISPR-Cas9系统设计并合成靶向PD-L1基因的sgRNA,构建PD-L1-PX458重组质粒并转染至SK-BR3中,利用WB检测筛选出基因编辑效率最高的质粒。随后依次使用CCK-8、transwell小室法及WB明确PD-L1对SK-BR3一般生物学功能的影响,并检测自噬相关蛋白LC3-Ⅱ/Ⅰ的表达水平。结果成功构建了能有效沉默SK-BR3中PD-L1基因的PD-L1-PX458重组质粒;PD-L1沉默显著抑制了SK-BR3的增殖活力(抑制率达36.7%),侵袭与迁移能力明显降低(P<0.05);DOX可诱导SK-BR3发生自噬,5μg/ml浓度作用下自噬程度最高,自噬水平与处理时间相关,以5μg/ml DOX处理SK-BR3,24 h时自噬水平最高;与SK-BR3野生株相比,DOX+PD-L1基因沉默组细胞中自噬水平明显增高(P<0.05)。结论PD-L1参与了SK-BR3的增殖与侵袭迁移,沉默PD-L1抑制DOX诱导的自噬水平,为乳腺癌的治疗提供了新的思路。Objective To explore the effect of programmed death-ligand 1(PD-L1)gene silencing on doxorubicin(DOX)-induced autophagy in breast cancer SK-BR3 cells.Methods The expression of PD-L1 in SK-BR3 cells was confirmed by Western blot(WB)and real-time quantitative PCR.The CRISPR-Cas9 system was used to design and synthesize sgRNA targeting the PD-L1 gene,and the PD-L1-PX458 recombinant plasmid was constructed and transfected into SK-BR3 cells.The plasmid with the highest gene editing efficiency was screened by WB.Subsequently,the CCK-8 assay,transwell chamber method,and WB were used to clarify the effect of PD-L1 on the general biological functions of SK-BR3 and to detect the expression levels of autophagy-related proteins LC3-Ⅱ/Ⅰ.Results The PD-L1-PX458 recombinant plasmid capable of effectively silencing the PD-L1 gene in SK-BR3 was successfully constructed.PD-L1 silencing significantly inhibited the proliferation activity of SK-BR3(inhibition rate of 36.7%),and the invasion and migration capabilities were significantly reduced(P<0.05).DOX can induce autophagy in SK-BR3,with the highest degree of autophagy at a concentration of 5μg/ml,and the level of autophagy was related to the treatment time,with the highest level of autophagy at 24 hours when treated with 5μg/ml DOX.Compared with the wild-type SK-BR3 cells,the level of autophagy in the DOX+PD-L1 gene-silenced group was significantly increased(P<0.05).Conclusions This study shows that PD-L1 is involved in the proliferation and invasion-migration of SK-BR3,and silencing PD-L1 inhibits DOX-induced autophagy levels,providing a new perspective for the treatment of breast cancer.

关 键 词：PD-L1基因 乳腺癌 多柔比星 自噬 

分 类 号：R737.9[医药卫生—肿瘤]