circCSPP1调控自噬影响胃癌细胞生长和转移能力的研究  

esearch of circCSPP1 affects the growth and metastasis ability of gastric cancer cells by regulating autophagy

作　　者：傅厚丰 李炳 梁振雄 王兴麟 FU Hou-feng;LI Bing;LIANG Zhen-xiong;WANG Xing-lin(Department of General Surgery,Qionghai People's Hospital,Qionghai 571400,Hainan Province,China)

机构地区：[1]琼海市人民医院普通外科,海南琼海571400

出　　处：《中国临床药理学杂志》2025年第5期665-670,共6页The Chinese Journal of Clinical Pharmacology

基　　金：海南省卫生健康行业科研基金资助项目(22A200336)。

摘　　要：目的探究circCSPP1介导的自噬对胃癌细胞增殖和转移能力的影响。方法将人胃癌细胞NCI-N87细胞分为Vector组(转染Vector)、circCSPP1组(转染circCSPP1)和抑制药组(转染circCSPP1后与10μmol·L^(-1)氯喹孵育)。将MKN-45细胞分为sh-NC组(转染circCSPP1 shRNA-NC)、sh-circCSPP1组(转染circCSPP1 shRNA)和激动药组(转染circCSPP1 shRNA后与1μmol·L^(-1)西罗莫司孵育)。用流式细胞术检测细胞的凋亡情况,用Transwell法检测细胞的迁移和侵袭情况,用蛋白质印迹法检测微管相关蛋白l轻链3-Ⅱ(LC3-Ⅱ)和p62蛋白的表达水平。结果在NCI-N87细胞中,Vector组、circCSPP1组和抑制药组的细胞凋亡率分别为(7.95±0.66)%、(3.93±0.62)%和(7.37±0.85)%,细胞迁移数分别为(122.50±12.25)、(169.40±16.09)和(109.00±8.22)个,细胞侵袭数分别为(73.38±12.33)、(117.30±12.78)和(55.25±8.73)个,LC3-Ⅱ蛋白相对表达水平分别为0.40±0.06、1.13±0.11和0.44±0.08,p62蛋白相对表达水平分别为1.62±0.15、0.21±0.08和1.43±0.16,抑制药组和Vector组的上述指标与circCSPP1组比较,在统计学上差异均有统计学意义(均P<0.05)。在MKN-45细胞中,sh-NC组、sh-circCSPP1组和激动药组的细胞凋亡率分别为(6.81±0.68)%、(12.02±0.78)%和(5.76±1.21)%,细胞迁移数分别为(191.60±21.43)、(87.50±10.76)和(155.60±10.36)个,细胞侵袭数分别为(150.00±11.55)、(61.63±11.39)和(154.50±17.99)个,LC3-Ⅱ蛋白相对表达水平分别为1.08±0.05、0.13±0.03和1.18±0.06,p62蛋白相对表达水平分别为0.15±0.03、1.29±0.08和0.19±0.04,激动药组和sh-NC组的上述指标与sh-circCSPP1组比较,在统计学上差异均有统计学意义(均P<0.05)。结论circCSPP1可以通过诱导自噬来提高胃癌细胞的增殖和转移能力。Objective To investigate the effects ofcircCSPP1 mediated autophagy on the proliferation and metastasis ability of gastric cancer cells.Methods The human gastric cancer cells NCI-N87 were divided into Vector group(transfected withVector),circCSPP1 group(transfected withcircCSPP1)and inhibitor group(transfected with circCSPP1 and incubated with 10μmol·L^(-1)chloroquine).The MKN-45 cells were divided into sh-NC group(transfected withcircCSPP1shRNA-NC),sh-circCSPP1 group(transfected withcircCSPP1shRNA)and agonist group(transfected withcircCSPP1 shRNA+1μmol·L^(-1)sirolimus).Flow cytometry was used to detect cell apoptosis.Transwell was used to detect cell migration and invasion.Western blot were used to detect the expression levels of microtubule associated protein light chain 3-Ⅱ(LC3-Ⅱ)and p62 protein.Results In NCI-N87 cells:the apoptosis rates of Vector,circCSPP1 and inhibitor groups were(7.95±0.66)%,(3.93±0.62)%and(7.37±0.85)%;the number of cell migrations were122.50±12.25,169.40±16.09 and 109.00±8.22;the number of cell invasions were 73.38±12.33,117.30±12.78 and 55.25±8.73;the relative expression levels of LC3-Ⅱprotein were 0.40±0.06,1.13±0.11and 0.44±0.08;the relative expression levels of p62 protein were 1.62±0.15,0.21±0.08 and 1.43±0.16.The differences of above indexes were statistically significant between the circCSPP1 group and the inhibitor and Vector groups(allP<0.05).In MKN-45 cells:the cell apoptosis rates of sh-NC,sh-circCSPP1 and agonist groups were(6.81±0.68)%,(12.02±0.78)%and(5.76±1.21)%;the number of cell migrations were 191.60±21.43,87.50±10.76 and 155.60±10.36;the number of cell invasions were 150.00±11.55,61.63±11.39 and154.50±17.99;the relative expression levels of LC3-Ⅱprotein were 1.08±0.05,0.13±0.03 and 1.18±0.06;the relative expression levels of p62 protein were 0.15±0.03,1.29±0.08 and 0.19±0.04.Compared with the sh-circCSPP1 group,the differences of above indicators in the agonist and sh-NC groups were statistically significant(allP<0.05).Conclusi

关 键 词：氯喹 西罗莫司 circCSPP1 胃癌 自噬 增殖 转移 

分 类 号：R28[医药卫生—中药学]