CAJ | 학술논문

目的：观察下调肾癌786-O细胞中叉头框转录因子M1（FoxM1）基因的表达对肾癌细胞生物学行为的影响。方法采用FoxM1的干扰序列和短发夹RNA（ shRNA）构建shFoxM1质粒。将786-O细胞分为shFoxM1组与对照组，分别转染shFoxM1、scramble质粒48 h。分别用Real-time PCR、Western blot法检测细胞FoxM1 mRNA及蛋白，平板克隆形成实验观察1周细胞克隆形成率，采用流式细胞仪检测细胞周期及细胞凋亡率，Transwell实验观察细胞迁移和侵袭能力，MTT法检测细胞培养0、24、48、72 h时的细胞吸光度值。结果与对照组比较，shFoxM1组FoxM1 mRNA和蛋白表达减少，细胞克隆形成率、细胞吸光度值降底，G1期细胞比例增加而G2、S期减少，早期凋亡率增加，迁移、侵袭的穿膜细胞减少，P均＜0．05。结论下调FoxM1基因表达，可抑制肾癌786-O细胞的增殖、迁移、侵袭能力，使细胞阻滞于G1期，促进细胞的早期凋亡。
목적：관찰하조신암786-O세포중차두광전록인자M1（FoxM1）기인적표체대신암세포생물학행위적영향。방법채용FoxM1적간우서렬화단발협RNA（ shRNA）구건shFoxM1질립。장786-O세포분위shFoxM1조여대조조，분별전염shFoxM1、scramble질립48 h。분별용Real-time PCR、Western blot법검측세포FoxM1 mRNA급단백，평판극륭형성실험관찰1주세포극륭형성솔，채용류식세포의검측세포주기급세포조망솔，Transwell실험관찰세포천이화침습능력，MTT법검측세포배양0、24、48、72 h시적세포흡광도치。결과여대조조비교，shFoxM1조FoxM1 mRNA화단백표체감소，세포극륭형성솔、세포흡광도치강저，G1기세포비례증가이G2、S기감소，조기조망솔증가，천이、침습적천막세포감소，P균＜0．05。결론하조FoxM1기인표체，가억제신암786-O세포적증식、천이、침습능력，사세포조체우G1기，촉진세포적조기조망。
Objective To observe the down-regulation of forkhead box protein M 1 ( FoxM1 ) gene expression and to investigate its effect on the biological behavior of human renal cancer cell line 786-O.Methods The shFoxM1 plasmid was constructed by RNA interference sequence and shRNA .786-O cells were divided into the shFoxM 1 group and the con-trol group which were transfected with shFoxM 1 plasmid and scramble plasmid for 48 h, respectively .The expression of FoxM1 mRNA and protein was tested by real-time PCR and Western blotting .Then we analyzed the efficiency of plate colo-ny formation in one week by using colony formation assay , and we used flow cytometry to detect the cell cycle and apoptosis of 786-O cells.The migration and invasion abilities were analyzed by Transwell assay , and the OD value in 0, 24, 48 and 72 hours was tested using MTT assay to analyze the activity of proliferation .Results Compared with the control group , the FoxM1 mRNA and protein expression was decreased , the efficiency of plate colony formation and the cell proliferation were decreased, the cells were blocked in G1 phase, cells of G2 phase and S phase reduced, the early apoptosis rate rose and the migration and invasion cells were decreased in the shFoxM 1 group (all P<0.05).Conclusions Down-regulating FoxM1 gene expression suppressed the proliferation , migration and invasion abilities of 786-O cells, blocking cells in G1 phase and promoting early apoptosis .