4-羟基水杨酰苯胺对T淋巴细胞白血病肿瘤增殖与凋亡的影响  

Effects of 4⁃hydroxysalicylaniline on the proliferation and apoptosis of T⁃cell lymphoblastic leukemia

作　　者：邓惠 陈格格 徐莉 贾新颜[2] 施菊妹 DENG Hui;CHEN Gege;XU Li;JIA Xinyan;SHI Jumei(Shanghai Clinical College,Anhui Medical University,Shanghai 200072;Department of Hematology,Shanghai East Hospital,Shanghai 200120;the Fifth Clinical College of Medicine,Anhui Medical University,Hefei 230032;Department of Hematology,Shanghai Tenth People’s Hospital Affiliated to Tongji University,Shanghai 200072,China)

机构地区：[1]安徽医科大学上海临床学院,上海200072 [2]上海市东方医院血液科,上海200120 [3]安徽医科大学第五临床医学院,安徽合肥230032 [4]同济大学附属上海第十人民医院血液科,上海200072

出　　处：《南京医科大学学报（自然科学版）》2023年第9期1185-1193,共9页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金(82170200,82170201,82170190)。

摘　　要：目的:研究4-羟基水杨酰苯胺(4-hydroxysalicylaniline,HDS)对T淋巴细胞白血病细胞Jurkat和Hut-78的抑制效果及其机制。方法:体外实验中,梯度浓度的HDS处理Jurkat和Hut-78细胞后,以CCK-8和克隆形成实验检测细胞增殖状况;用细胞流式术检测细胞周期变化,通过Western blot检测周期相关蛋白的表达水平;用细胞流式术检测细胞凋亡水平,通过Western blot检测凋亡相关蛋白的表达水平;以γ-H2AX免疫荧光染色检测DNA损伤状况,并通过Western blot检测DNA损伤相关蛋白的表达水平。体内实验中,通过皮下注射Jurkat细胞构建小鼠肿瘤模型,处理组腹腔注射HDS,对照组给予等量溶剂,记录小鼠体重与肿瘤体积变化,13 d后,取肿瘤组织,通过HE染色观察组织形态,并通过Ki67和γ-H2AX的免疫组织化学染色,分析药物对肿瘤组织增殖与DNA损伤的影响。结果:在体内和体外,证明了HDS可通过诱导Jurkat和Hut-78细胞S期细胞周期阻滞与凋亡,并且通过CHK1/CHK2信号通路的磷酸化激活,加重DNA损伤,抑制T淋巴细胞白血病肿瘤,但HDS对小鼠无明显毒性。结论:HDS可作为潜在的抗T淋巴细胞白血病药物,可抑制细胞周期,诱导细胞凋亡,并通过CHK1/CHK2信号通路影响DNA损伤修复过程。Objective:The current study aims to investigate the effect of 4⁃hydroxysalicylaniline(HDS)on the proliferation and apoptosis of T⁃lymphoblastic leukemia cells,Jurkat and Hut⁃78,in vitro and in vivo.Methods:In vitro,HDS was used to treat Jurkat and Hut⁃78 cells with different concentration gradients.The CCK⁃8 assay and colony formation assay were used to detect the effect of HDS on cell proliferation.Cell cycle changes were detected by flow cytometry,and the expression levels of cycle⁃related proteins were detected by Western blot.The apoptosis level was detected by flow cytometry,and the expression levels of apoptosis⁃related proteins were detected by Western blot.DNA damage was detected byγ⁃H2AX immunofluorescence staining,and the expression levels of DNA damage related proteins were detected by Western blot.In vivo,the mouse T⁃cell lymphoblastic leukemia tumor model was constructed by subcutaneous injection of Jurkat cells,and HDS was injected to observe the effect of drugs on tumor volume and body weight of mice.HE staining was used to observe the morphological changes of tumors after 13 days of HDS creatment.Immunohistochemical staining of Ki67 andγ⁃H2AX protein was used to analyze the effect of drugs on cell proliferation and DNA damage in tumors.Results:In Jurkat and Hut⁃78 cell lines,HDS could induce cell cycle arrest at S phase,induce cell apoptosis,and block DNA damage repair through phosphorylation of CHK1/CHK2 signaling pathway,both in vitro and in vivo.HDS inhibited T⁃cell lymphoblastic leukemia tumors,and it had no obvious toxicity to mice.Conclusion:HDS can be used as a potential anti⁃T⁃cell lymphoblastic leukemia drug.The effect of HDS on T⁃cell lymphoblastic leukemia may be accomplished by interfering cell cycle and apoptosis,and DNA damage repair through CHK1/CHK2 signaling pathway.

关 键 词：T淋巴细胞白血病 4-羟基水杨酰苯胺 细胞周期S期阻滞 细胞凋亡 DNA损伤 

分 类 号：R733.71[医药卫生—肿瘤]