安徽医科大学学报
安徽醫科大學學報
안휘의과대학학보
ACTA UNIVERSITY MEDICINALIS ANHUI
2014年
6期
748-751
,共4页
陈杰勋%于德新%张志强%王铮%谢栋栋%王毅%张涛
陳傑勛%于德新%張誌彊%王錚%謝棟棟%王毅%張濤
진걸훈%우덕신%장지강%왕쟁%사동동%왕의%장도
RNA干扰%Eg5基因%膀胱癌
RNA榦擾%Eg5基因%膀胱癌
RNA간우%Eg5기인%방광암
RNA interference%Eg5 gene%bladder carcinoma
目的：通过RNA干扰阻断T24细胞中Eg5基因的表达,研究Eg5基因对 T24细胞增殖和迁移的影响。方法T24细胞株分为3组：干扰组、空白对照组、阴性对照组。化学合成针对Eg5的小干扰RNA,通过脂质体转染至T24细胞中,应用Western blot法检测转染后T24细胞中Eg5蛋白表达,采用台盼蓝拒染实验检测细胞死亡率,用噻唑蓝( MTT)比色分析法和平板克隆形成实验检测细胞增殖抑制率,通过细胞划痕实验检测细胞的迁移能力。结果干扰组Eg5蛋白表达显著低于空白对照组和阴性对照组；干扰组细胞死亡率显著升高,与空白对照组及阴性对照组比较差异有统计学意义( P<0．05)；干扰组的克隆形成率下降,与空白对照组和阴性对照组比较差异有统计学意义( P <0．05)；MTT结果提示,干扰Eg5基因表达后,T24细胞的增殖率明显下降(P<0．05)；划痕实验结果显示干扰组细胞迁移能力受到明显抑制(P<0．05)。结论 Eg5基因在膀胱移行细胞癌起重要作用,干扰Eg5基因能有效抑制T24细胞的增殖与迁移,促进细胞死亡,以Eg5为靶点有望成为膀胱癌基因治疗的新基因靶点。
目的：通過RNA榦擾阻斷T24細胞中Eg5基因的錶達,研究Eg5基因對 T24細胞增殖和遷移的影響。方法T24細胞株分為3組：榦擾組、空白對照組、陰性對照組。化學閤成針對Eg5的小榦擾RNA,通過脂質體轉染至T24細胞中,應用Western blot法檢測轉染後T24細胞中Eg5蛋白錶達,採用檯盼藍拒染實驗檢測細胞死亡率,用噻唑藍( MTT)比色分析法和平闆剋隆形成實驗檢測細胞增殖抑製率,通過細胞劃痕實驗檢測細胞的遷移能力。結果榦擾組Eg5蛋白錶達顯著低于空白對照組和陰性對照組；榦擾組細胞死亡率顯著升高,與空白對照組及陰性對照組比較差異有統計學意義( P<0．05)；榦擾組的剋隆形成率下降,與空白對照組和陰性對照組比較差異有統計學意義( P <0．05)；MTT結果提示,榦擾Eg5基因錶達後,T24細胞的增殖率明顯下降(P<0．05)；劃痕實驗結果顯示榦擾組細胞遷移能力受到明顯抑製(P<0．05)。結論 Eg5基因在膀胱移行細胞癌起重要作用,榦擾Eg5基因能有效抑製T24細胞的增殖與遷移,促進細胞死亡,以Eg5為靶點有望成為膀胱癌基因治療的新基因靶點。
목적：통과RNA간우조단T24세포중Eg5기인적표체,연구Eg5기인대 T24세포증식화천이적영향。방법T24세포주분위3조：간우조、공백대조조、음성대조조。화학합성침대Eg5적소간우RNA,통과지질체전염지T24세포중,응용Western blot법검측전염후T24세포중Eg5단백표체,채용태반람거염실험검측세포사망솔,용새서람( MTT)비색분석법화평판극륭형성실험검측세포증식억제솔,통과세포화흔실험검측세포적천이능력。결과간우조Eg5단백표체현저저우공백대조조화음성대조조；간우조세포사망솔현저승고,여공백대조조급음성대조조비교차이유통계학의의( P<0．05)；간우조적극륭형성솔하강,여공백대조조화음성대조조비교차이유통계학의의( P <0．05)；MTT결과제시,간우Eg5기인표체후,T24세포적증식솔명현하강(P<0．05)；화흔실험결과현시간우조세포천이능력수도명현억제(P<0．05)。결론 Eg5기인재방광이행세포암기중요작용,간우Eg5기인능유효억제T24세포적증식여천이,촉진세포사망,이Eg5위파점유망성위방광암기인치료적신기인파점。
Objective To observe the bionomics effects of Eg5 gene on human T24 tumor cell line. Methods T24 cells were divided into three groups:the Eg5-siRNA group,the blank control group,the negative group. The expres-sion of Eg5 was detected by Western blot. The rate of cell death was determined by Tyrpan blue staining assays. The proliferation of T24 cells was determined by methyl thiazol tetrazolium( MTT) assay and plate clone formation assay. The migration of T24 cells was analyzed by scratch test. Results Western blot analysis showed that T24 cells with Eg5-siRNA transfected decreased the Eg5 gene expression. In contrast to the control group, Eg5-siRNA induced markedly the T24 cellls death. The ability of proliferation and migration of T24 cells treated with Eg5-siRNA was decreased as compared with the blank control group and the negative group(P<0. 05). Conclusion Silencing Eg5 can regulate the malignant biological behavirors of bladder cancer cell line T24 effectively and may become a novel gene therapeutic strategy for bladder cancer.
