甘草查尔酮A对HL-60细胞恶性生物学行为的抑制作用及其机制  

Inhibitory effect of licochalcone A on malignant biological behavior of HL-60 cells and its mechanism

作　　者：韩萍萍[1] 王怀宇[2] 蔡云[1] 孙烨[1] 夏欣欣[1] 胡珊 周冬枝[1] 韦素华 HAN Pingping;WANG Huaiyu;CAI Yun;SUN Ye;XIA Xinxin;HU Shan;ZHOU Dongzhi;WEI Suhua(Department of Traditional Chinese Medicine,First Affiliated Hospital of Xi′an Jiaotong University,Xi′an 710061,China;Department of Hematology,First Affiliated Hospital of Xi′an Jiaotong University)

机构地区：[1]西安交通大学第一附属医院中医科,西安710061 [2]西安交通大学第一附属医院血液科

出　　处：《山西医科大学学报》2024年第6期689-697,共9页Journal of Shanxi Medical University

基　　金：陕西省重点研发计划项目(2024SF-YBXM-521);陕西省中医药管理局委托办事经费项目(ZYJXG-L23020)。

摘　　要：目的 探究甘草查尔酮A(LA)对急性髓系白血病(AML)细胞HL-60的作用及其机制。方法 将人AML细胞系HL-60细胞分为对照组、LA组(30μmol/L)、IL-6组(100 ng/mL,JAK2/STAT3信号通路激活剂)和LA+IL-6组。培养48 h后,采用MTT法和克隆形成实验检测HL-60细胞增殖活性;采用流式细胞术检测HL-60细胞周期分布和细胞凋亡;采用Transwell实验检测HL-60细胞侵袭和迁移;采用Western blot检测p21、p27、cyclin E2、CDK2、Bcl-2、Bax、cleaved-Caspase-3、p-JAK2,JAK2,p-STAT3和STAT3蛋白表达水平。结果 与对照组比较,LA组细胞活力、克隆形成数、侵袭和迁移细胞数均降低(P<0.05),Bcl-2、cyclin E2和CDK2蛋白表达量均降低(P<0.05),JAK2和STAT3蛋白的磷酸化水平均降低(P<0.05);G0/G1期细胞比例和细胞凋亡率均升高(P<0.05),p21、p27、Bax和cleaved Caspase-3蛋白表达量均升高(P<0.05)。与对照组比较,IL-6组细胞活力、克隆形成数、侵袭和迁移细胞数均升高(P<0.05),Bcl-2、cyclin E2和CDK2蛋白表达量均升高(P<0.05),JAK2和STAT3蛋白的磷酸化水平均升高(P<0.05),G0/G1期细胞比例和细胞凋亡率均降低(P<0.05),p21、p27、Bax和cleaved Caspase-3蛋白表达量均降低(P<0.05)。与IL-6组比较,LA+IL-6组细胞活力、克隆形成数、侵袭和迁移细胞数均降低(P<0.05),Bcl-2、cyclin E2和CDK2蛋白表达量均降低(P<0.05),JAK2和STAT3蛋白的磷酸化水平均降低(P<0.05),G0/G1期细胞比例和细胞凋亡率均升高(P<0.05),p21、p27、Bax和cleaved-Caspase-3蛋白表达量均升高(P<0.05)。LA+IL-6组和LA组间上述指标差异没有统计学意义。结论 甘草查尔酮A可抑制AML细胞系HL-60细胞增殖、侵袭和迁移,诱导细胞凋亡并将细胞阻滞在G0/G1期,其作用机制可能与抑制JAK2/STAT3信号通路激活有关。Objective To explore the effect of licochalcone A(LA)on acute myeloid leukemia(AML)HL-60 cell and its mechanism.Methods Human AML cell line HL-60 cells were divided into control group,LA group(30μmol/L LA),IL-6 group(100 ng/mL IL-6,an activator of JAK2/STAT3 signaling pathway)and LA+IL-6 group.After 48 h culture,the proliferation activity of HL-60 cells was detected by MTT assay and clonal formation assay.The cell cycle distribution and the apoptosis of HL-60 cells were detected by flow cytometry.The invasion and migration abilities of HL-60 cells were detected by Transwell assay.The protein expression levels of p21,p27,cyclin E2,CDK2,Bcl-2,Bax,cleaved Caspase-3,p-JAK2,JAK2,p-STAT3 and STAT3 were detected by Western blot.Results Compared with control group,the cell viability,the number of clonal formation,the numbers of invasion and migration cells were decreased in LA group(P<0.05),the protein expression levels of Bcl-2,cyclin E2 and CDK2 were decreased(P<0.05),the phosphorylation levels of JAK2 and STAT3 were decreased(P<0.05),the proportion of cells in G0/G1 phase and the apoptosis rate were increased(P<0.05),and the protein expression levels of p21,p27,Bax and cleaved Caspase-3 were increased(P<0.05).Compared with control group,the cell viability,the number of clonal formation,the numbers of invasion and migration cells were increased in IL-6 group(P<0.05),the protein expression levels of Bcl-2,cyclin E2 and CDK2 were increased(P<0.05),the phosphorylation levels of JAK2 and STAT3 were increased(P<0.05),the proportion of cells in G0/G1 phase and the apoptosis rate were decreased(P<0.05),and the protein expression levels of p21,p27,Bax and cleaved Caspase-3 were decreased(P<0.05).Compared with IL-6 group,the cell viability,the number of clonal formation,the numbers of invasion and migration cells were decreased in LA+IL-6 group(P<0.05),the protein expression levels of Bcl-2,cyclin E2 and CDK2 were decreased(P<0.05),the phosphorylation levels of JAK2 and STAT3 were decreased(P<0.05),the proportion of cells in G

关 键 词：甘草查尔酮A 急性髓系白血病 生物学行为 JAK2/STAT3信号通路 HL-60细胞 

分 类 号：R733.71[医药卫生—肿瘤]