长链非编码RNA CASC2调节HepG2肝癌细胞的炎症因子分泌并诱导细胞凋亡和生长阻滞  

Long non coding RNA CASC2 regulates inflammatory cytokine secretion and induces apoptosis and growth arrest of HepG2 cells

作　　者：郝剑[1] 苏华[2] 贺杰[1] 马俊帅[2] 韩磊[3] HAO Jian;SU Hua;HE Jie;MA Junshuai;HAN Lei(Department of Blood Transfusion,First Affiliated Hospital of Hebei North University,Zhangjiakou 075000,China;Department of Laboratory Medicine,First Affiliated Hospital of Hebei North University,Zhangjiakou 075000,China;Department of General Surgery,First Affiliated Hospital of Hebei North University,Zhangjiakou 075000,China)

机构地区：[1]河北北方学院附属第一医院输血科,075061 [2]河北北方学院附属第一医院检验科,075061 [3]河北北方学院附属第一医院普外科,075061

出　　处：《免疫学杂志》2021年第11期947-953,共7页Immunological Journal

基　　金：河北省医学科学研究重点课题(12110051D)。

摘　　要：目的研究长链非编码RNA癌症易感候选物2(CASC2)对肝癌细胞肿瘤免疫微环境的调节和对细胞增殖、凋亡的影响。方法 RT-qPCR检测肝癌组织和细胞中LncRNA CASC2的表达。运用Lipofectamine 2000将LV-scramble和LV-CASC2质粒分别转染到肝癌HepG2细胞。ELISA检测细胞上清液中IL-6、IL-10、IL-4和IL-1β的含量,评估肝癌细胞肿瘤免疫微环境;克隆形成实验检测细胞增殖;流式细胞术检测细胞凋亡;Western blot检测细胞裂解液中p65的磷酸化和增殖相关蛋白p21,Ki67和Survivin的表达。结果 LncRNA CASC2在肝癌组织中下调(P<0.05);与Control组相比,LV-CASC2转染组HepG2细胞中IL-6和IL-1β水平显著升高,IL-10和IL-4水平显著降低,克隆形成能力降低,细胞凋亡率升高(P<0.05)。结论 LncRNA CASC2可通过改变HepG2细胞炎症因子的分泌、阻止细胞增殖和诱导细胞凋亡来实现肝癌肿瘤的生长阻滞。To study the regulation of long-chain non-coding RNA cancer susceptibility candidate 2(CASC2) on the tumor immune microenvironment of liver cancer cells and its effect on cell proliferation and apoptosis, RTqPCR was used to detec the expression of LncRNA CASC2 in liver cancer tissues and cells. Lipofectamine 2000 was used to transfect LV-scramble and LV-CASC2 plasmids into HepG2 cells;ELISA was used to detect the contents of IL-6, IL-10, IL-4 and IL-1β in the cell supernatant for evaluating the tumor immune microenvironment of liver cancer cells;clone formation test was performed to detect cell proliferation, while flow cytometry was used to detecte cell apoptosis;Western blot was applied to detect the phosphorylation of p65 and the expression of proliferationrelated proteins p21, Ki67 and survivin in the cell lysate. Data showed that LncRNA CASC2 was down-regulated in liver cancer tissues(P<0.05). Compared with the control group, the levels of IL-6 and IL-1β in HepG2 cells of the LV-CASC2 transfection group were significantly increased, the levels of IL-10 and IL-4 were significantly reduced,the cloning ability was reduced, while the apoptosis rate was increased(P<0.05). In conclusion, lncRNA CASC2 could induce the growth arrest of liver cancer tumors by destroying the inflammatory microenvironment of liver cancer cells, preventing cell proliferation and inducing cell apoptosis.

关 键 词：LncRNA CASC2 HEPG2细胞 肝癌 P21 炎性微环境 

分 类 号：R735.7[医药卫生—肿瘤]