摘要
目的观察胶质瘤相关癌基因同源物(glioma-associated oncogene homolog,Gli)通过Polo样激酶1(Polo-like kinase 1,PLK1)对食管腺癌肿瘤生物学行为的影响并探讨其相关的分子机制。方法应用二甲亚砜(dimethyl sulfoxide,DMSO)、GANT61、N-Shh蛋白及GANT61&N-Shh分别处理食管腺癌OE33细胞株24 h后,实时荧光定量PCR法和Western blot法观察Gli1、Gli2和PLK1的表达;GANT61处理细胞48 h后观察对Gli1、Gli2、PLK1蛋白表达的影响,流式细胞术观察GANT61处理细胞72 h后对细胞周期和凋亡的影响。结果 GANT61组在Gli1 mRNA和Gli1蛋白、Gli2 mRNA和Gli1蛋白以及PLK1 mRNA和Gli1蛋白的表达明显低于DMSO组,N-Shh组明显高于DMSO组和GANT61组,GANT61+N-Shh组低于DMSO组,但高于GANT61组(P<0.05);GANT61组G0/G1期、S期和G2/M期增殖率以及GANT61组培养72 h凋亡率均高于DMSO组(P<0.05)。结论 Gli和PLK1异常表达在食管腺癌细胞中存在串活机制,两者可以调节细胞周期和凋亡,在食管腺癌发生发展中发挥重要作用。
Objective To explore the effect Gli expression on growth of esophageal adenocarcinoma cells through PLK1 and to detect their molecular mechanisms.Methods mRNA and protein expression of Gli1,Gli2 and PLK1 were detected after 24 h treatment of DMSO,GANT61,N-Shh and GANT61&N-Shh respective by real time fluorescence quantitative PCR and Western blot assay.Cell cycle and apoptosis were measured by flow cytometry after 72 h treatment of GANT61.Results Compared with DMSO group,real-time fiuorescent quantitative PCR and Western blotting showed that GANT61 could downregulate mRNA and protein expression of Gli1,Gli2and PLK1,while N-Shh could increase mRNA and protein expression of Gli1,Gli2and PLK1 significantly.GANT61&N-Shh could restore partly mRNA and protein expression of Gli1,Gli2 and PLK1.OE33 cells showed a strong G2/M arrest and apoptosis 72 h after treatment of GANT61(P<0.01).Expression of Gli1 mRNA and Gli1 protein,Gli2 mRNA and Gli1 protein,PLK1 mRNA and Gli1 in GANT61 group were lower than those in DMSO group.Those indexes were higher in N-shh group than in DMSO group and GANT61 group,GANT61+N-Shh group were lower than DMSO group,but higher than GANT61 group.Proliferation rate of G0/G1 stage,S stage and G2/M stage in GANT61 group were higher than DMSO group,and it was the same trend with 72 h apoptotic rate between GANT61 group and DMSO group.Conclusion Glihas crosstalk relationship with PLK1 which may regulate cell cycle and apoptosis.They may play orchestral role in the growth of esophageal adenocarcinoma.
作者
赵鑫
杜媛鲲
张耀中
张磊
米源
王雷
ZHAO Xin;DU Yuan-kun;ZHANG Yao-zhong;ZHANG Lei;MI Yuan;WANG Lei(Department of Function Inspection,Hebei General Hospital,Shijiazhuang 050051,China;Agency of Periodicals,Hebei Medical University,Shijiazhuang 050017,China;Department of Thoracic Surgery,the Fourth Hospital of HeBei Medical University,Shijiazhuang 050011,China;Department of Surgery,Xingtai County Central Hospital,Heibe Province,Xingtai 054001,China)
出处
《河北医科大学学报》
CAS
2019年第5期528-532,共5页
Journal of Hebei Medical University
基金
河北省医学科学研究重点课题(20180528)
河北省留学人员科技活动项目择优资助(CY201613)
