摘要
目的探讨着丝粒蛋白U(CENPU)对胃癌细胞增殖、糖酵解的影响及可能机制。方法体外培养人胃癌细胞系GES-1、AGS、MKN45、HGC-27细胞,采用实时荧光定量PCR法测定各细胞系中CENPU mRNA表达;根据CENPU mRNA表达情况筛选HGC-27细胞进行后续实验,并将其分为空白对照组(NG组:细胞不做特殊处理)、阴性对照组(si-CENPU-NC组:转染si-CENPU-NC序列)、沉默CENPU表达组(si-CENPU组:转染si-CENPU序列)。采用CCK-8法、平板克隆实验分别检测各组HGC-27细胞增殖情况、克隆形成率;酶联免疫吸附(ELISA)法测定各组HGC-27细胞糖酵解过程关键指标葡萄糖消耗(GC)、乳酸脱氢酶(LDH)、三磷酸腺苷(ATP)、丙酮酸激酶(PK)、己糖激酶(HK)水平;免疫印迹法检测各组HGC-27细胞蛋白激酶B(AKT)、p-AKT、转录激活因子3(STAT3)、p-STAT3蛋白表达。结果CENPU mRNA在胃癌细胞中异常高表达,且HGC-27细胞中CENPU mRNA表达最高(P<0.05),选择HGC-27细胞进行后续实验。与NG组、si-CENPU-NC组比较,si-CENPU组HGC-27细胞增殖抑制率升高(P均<0.05),克隆形成率、细胞中GC、LDH、ATP、HK、PK水平及p-AKT/AKT、p-STAT3/STAT3水平均降低(P均<0.05)。结论下调CENPU表达可抑制胃癌细胞增殖并干扰细胞糖酵解,其机制可能通过抑制AKT/STAT3通路活化实现。
Objective To investigate the effects and possible mechanism of centromere protein U(CENPU)on the proliferation and glycolysis of gastric cancer cells.Methods Human gastric cancer cell lines GES-1,AGS,MKN45,and HGC-27 were cultured in vitro,and the expression of CENPU mRNA in each cell line was determined by real-time quantitative PCR.HGC-27 cells were screened for subsequent experiments according to the expression of CENPU mRNA.They were divided into the blank control group(NG group:cells without special treatment),negative control group(siCENPU-NC group:transfected with si-CENPU-NC sequence),and silent CENPU expression group(si-CENPU group:transfected with si-CENPU sequence).The proliferation and clone formation rate of HGC-27 cells in each group were de⁃tected by CCK-8 and plate cloning experiment.Enzyme-linked immunosorbent assay(ELISA)was used to determine the key indicators of glycolysis in HGC-27 cells in each group:glucose consumption(GC),lactate dehydrogenase(LDH),ad⁃enosine triphosphate(ATP),and pyruvate kinase(PK),hexokinase(HK)levels.Western blotting was used to detect protein kinase B(AKT),p-AKT,activator of transcription 3(STAT3)and p-STAT3 protein expression in HGC-27 cells in each group.Results The expression of CENPU mRNA was abnormally high in gastric cancer cells,and the expres⁃sion of CENPU mRNA was the highest in HGC-27 cells(P<0.05).HGC-27 cells were selected for subsequent experi⁃ments.Compared with the NG group and si-CENPU-NC group,the proliferation inhibition rate of HGC-27 cells increased(P<0.05),and the colony formation rate,the levels of GC,LDH,ATP,HK,PK and p-AKT/AKT,and p-STAT3/STAT3 in cells all decreased in the si-CENPU group(all P<0.05).Conclusion Down-regulation of CENPU expression can inhibit the proliferation of gastric cancer cells and interfere with cellular glycolysis,which may be achieved by inhibit⁃ing the activation of AKT/STAT3 pathway.
作者
邓桃枝
江雪梅
何周桃
蔡曼妮
陈超超
许泽文
DENG Taozhi;JIANG Xuemei;HE Zhoutao;CAI Manni;CHEN Chaochao;XU Zewen(Department of Gastroenterology,Hainan Cancer Hospital,Haikou 570312,China)
出处
《山东医药》
CAS
2022年第10期48-51,共4页
Shandong Medical Journal
关键词
胃癌
着丝粒蛋白U
细胞增殖
糖酵解
gastric carcinoma
centromeric protein U(CENPU)
cell proliferation
glycolysis
