siRNA沉默PTTG表达对子宫内膜癌细胞生长及放疗敏感性的影响  

Effects of silencing PTTG expression by small interference RNA on proliferation and radiotherapeutic sensitivity of endometrial carcinoma

作　　者：张曙萱[1,2] 袁丽萍[2] 刘永利[2] 孙丽洲[1] 

机构地区：[1]南京医科大学第一附属医院妇产科,江苏南京210029 [2]徐州市第一人民医院妇产科,江苏徐州221002

出　　处：《南京医科大学学报（自然科学版）》2012年第11期1527-1532,共6页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金(81070511)

摘　　要：目的:观察垂体瘤转化基因(pituitary tumor transforming gene,PTTG)小干扰RNA(small interference RNA,siRNA)干扰对子宫内膜癌细胞HEC-1A的生长及放疗敏感性的影响。方法:构建PTTG干扰载体(pSilencer3.0-Hl-PTTG-siRNA),利用脂质体将其转染至HEC-1A细胞,Western blot检测PTTG蛋白表达水平,筛选干扰效果最强的序列。另用2Gy X线作用于转染PTTGsiRNA的HEC-1A细胞,同时设不予任何处理的对照组、只转染PTTGsiRNA的处理组、只用X线处理的放疗组。MTT检测细胞生长状况,流式细胞仪检测细胞凋亡。结果:成功构建了靶向抑制PTTG基因的siRNA干扰载体。MTT实验结果显示,与对照组相比,放疗组、PTTGsiRNA转染组、PTTGsiRNA联合放疗组细胞增殖抑制率分别为(31.39±5.62)%、(38.37±5.48)%、(54.65±6.27)%。流式细胞仪检测显示细胞凋亡率在对照组、放疗组、PTTGsiRNA转染组、PTTGsiRNA转染联合放疗组中分别为(6.53±0.80)%、(32.72±4.56)%、(38.96±4.37)%、(64.76±6.53)%。PTTGsiRNA转染组或放疗组与对照组比较,细胞凋亡率及细胞增殖抑制率差异均有统计学意义(P<0.01),而PTTGsiRNA转染联合放疗组的凋亡率及细胞增殖抑制率较PTTGsiRNA转染组或放疗组明显增高,组间比较差异均有统计学意义(P<0.01)。结论:成功构建了靶向抑制PTTG基因的siRNA干扰载体,PTTG siRNA可抑制子宫内膜癌细胞HEC-1A的增殖,诱导其凋亡,增强其对放疗的敏感性。Objective：To investigate the effects of silence PTTG expression by small interference RNA（siRNA） on proliferation and radiotherapeutic sensitivity of endometrial carcinoma cells（HEC-1A）.Methods：Vectors pSilencer3.0-H1-PTTG-SiRNA were constructed to transcribe functional siRNA specially targeting PTTG.The interfering plasmids were used to transfect HEC-1A cells with lipofectmine 2000 transfection reagent.PTTG protein expression levels were analyzed by Western blot method.The HEC-1A cells were treated with PTTG siRNA,radiotherapy and PTTG siRNA combined with radiotherapy,and cells without transfection were as control.MTT assay was used to detected cell proliferation,and the apoptosis of HEC-1A cells was evaluated by flow cytometry.Results：Recombinant siRNA expression vector targeting PTTG was constructed.The result of MTT showed that the growth of HEC-1A cells was influenced after PTTGsiRNA transfection,radiotherapy and PTTGsiRNA transfection combined with radiotherapy.Inhibitory rate of radiotherapy group,PTTGsiRNA group and PTTGsiRNA combined with radiotherapy group were（31.39 ± 5.62）%,（38.37 ± 4.48）%,（54.65 ± 6.27）%,respectively.The apoptosis rate at 48 h of control group,radiotherapy group,PTTGsiRNA group,PTTGsiRNA combined with radiotherapy group were（6.53 ± 0.80）%,（32.72 ± 4.56）%,（38.96 ± 4.37）%,（64.76 ± 6.53）%,respectively,by the flow cytometry analysis.There were significant differences between control group and PTTGsiRNA group or radiotherapy group,and there were significant differences between PTTGsiRNA combined with radiotherapeutic group and PTTGsiRNA group or radiotherapy group.Conclusion：The siRNA expression vector targeting PTTG was successfully constructed.PTTG RNA interference may inhibit the cell proliferation and induce apoptosis in endometrial carcinoma cells,which can upregulate radiotherapeutic sensitivity of HEC-1A cells.

关 键 词：SIRNA 子宫内膜癌 垂体瘤转化基因 放疗敏感性 

分 类 号：R737.33[医药卫生—肿瘤]