白细胞介素-37对HepG2肝癌细胞增殖、凋亡和周期的影响  

Effects of interleukin-37 on proliferation,apoptosis and cycle of HepG2 hepatoma cells

作　　者：方萍 蔡德成 孟萍 王蓉 韩秀娟 张成芳[1] FANG Ping;CAI Decheng;MENG Ping;WANG Rong;HAN Xiujuan;ZHANG Chengfang(Department of Medical Laboratory,Huadu District People's Hospital of Guangzhou,Guangzhou 510800,China;Department of Obstetrics and Gynecology,The Third Affiliated Hospital of Southern Medical University,Guangzhou 510000,China;Central Laboratory,Huadu District People's Hospital of Guangzhou,Guangzhou 510800,China)

机构地区：[1]广州市花都区人民医院医学检验科,广州510800 [2]南方医科大学第三附属医院妇产科,广州510000 [3]广州市花都区人民医院中心实验室,广州510800

出　　处：《数理医药学杂志》2025年第3期178-185,共8页Journal of Mathematical Medicine

基　　金：广州市花都区科技计划项目(22-HDWS-034);广州市花都区人民医院院内科研基金项目(2020C05)。

摘　　要：目的探究白细胞介素-37(interleukin-37,IL-37)炎症抑制因子对HepG2肝癌细胞增殖、凋亡和周期的影响。方法将IL-37的慢病毒表达载体pCDH-CMV-MCSIL37-copGFP-T2A-Puro(pCDH-IL-37组)与空白载体pCDH-CMV-MCS-EF1-copGFPT2A-Puro(pCDH组)分为实验组与对照组,分别对两组进行病毒包装与嘌呤霉素筛选,然后转染入体外培养的HepG2细胞。通过实时荧光定量逆转录聚合酶链反应(quantitative real-time reverse transcription polymerase chain reaction,RT-qPCR)检测转染后的HepG2细胞中的IL-37 mRNA表达水平。通过细胞计数试剂盒-8(cell counting kit-8,CCK-8)法检测IL-37对HepG2细胞增殖的影响。通过流式细胞术检测IL-37对HepG2细胞周期及凋亡的影响。结果经过慢病毒包装、感染和嘌呤霉素筛选,荧光显微镜显示慢病毒表达载体成功转染到HepG2细胞中。RT-qPCR结果显示,HepG2细胞中IL-37相对表达水平高。CCK-8法检测结果显示,细胞培养24 h、48 h、72 h后,与pCDH组比较,pCDH-IL-37组细胞450 nm波长处的光密度(optical density,OD),即OD450,吸光值明显降低,差异具有统计学意义(P<0.001),且随着培养时间的延长,细胞存活率逐渐降低。流式细胞术检测细胞周期结果显示,与pCDH组比较,pCDH-IL-37组的S期细胞比率显著降低,G1期细胞比率显著升高,细胞周期被阻滞于G1期。流式细胞术检测细胞凋亡结果显示,pCDHIL-37组的细胞凋亡率显著高于pCDH组(9.833%±0.252%vs.4.867%±0.569%,P<0.001)。RT-qPCR结果显示,pCDH-IL-37组Caspase-3与Fas的mRNA表达水平降低,表明IL-37对Caspase-3与Fas诱导的凋亡可能具有抵抗作用。结论IL-37在体外可以直接抑制肝癌细胞HepG2的增殖,促进其凋亡,将细胞周期阻滞在G1期,并可能通过下调Caspase-3与Fas的表达,对Caspase-3与Fas诱导的凋亡发挥抵抗作用。Objective To investigate the effect of interleukin-37(IL-37)inflammatory inhibitor on the proliferation,apoptosis and cycle of HepG2 hepatoma cells.Methods The lentiviral expression vector pCDH-CMV-MCS-IL37-copGFP-T2A-Puro with the IL-37(pCDH-IL-37 group)and blank vector pCDHCMV-MCS-EF1-copGFP-T2A-Puro(pCDH group)were divided into the experimental group and the control group,respectively.The two groups were screened for viral packaging and puromycin,respectively,and then transfected into HepG2 cells in vitro.The mRNA expression level of IL-37 in transfected HepG2 cells was determined by quantitative real-time reverse transcription polymerase chain reaction(RT-qPCR).The effect of IL-37 on the proliferation of HepG2 cells was detected by cell counting kit-8(CCK-8).The effects of IL-37 on the cycle and apoptosis of HepG2 cells were evaluated by flow cytometry.Results After lentivirus packaging,infection,and puromycin screening,fluorescence microscopy showed that the lentiviral expression vectors were successfully transfected into HepG2 cells.RT-qPCR showed a high level of IL-37 expression in HepG2 cells.The results of CCK-8 assay showed that after 24 h,48 h and 72 h of cell culture,the optical density(OD)at 450 nm of cells in PCDH-IL-37 group was significantly lower than that in pCDH group.The difference was statistically significant(P<0.001),and the cell survival rate decreased gradually with the extension of culture time.Flow cytometry showed that compared with pCDH group,the ratio of S phase cells was significantly decreased in PCDH-IL-37 group,and the ratio of G1 phase cells was significantly increased,and the cell cycle was blocked in G1 phase.Flow cytometry showed that the apoptosis rate of PCDH-IL-37 group was significantly higher than that of pCDH group(9.833%±0.252%vs.4.867%±0.569%,P<0.001).RT-qPCR results showed that mRNA expression levels of Caspase-3 and Fas were decreased in pCDH-IL-37 group,suggesting that IL-37 may have a resistance to Caspase-3 and Fas induced apoptosis.Conclusion IL-37 can directl

关 键 词：白细胞介素-37 肝癌 HEPG2细胞 细胞增殖 细胞凋亡 细胞周期 

分 类 号：R735.7[医药卫生—肿瘤]