中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
CHINESE JOURNAL OF PATHOPHYSIOLOGY
2015年
5期
857-863
,共7页
巫翟%夏忠胜%钟娃%卢锡基%于涛%陈其奎
巫翟%夏忠勝%鐘娃%盧錫基%于濤%陳其奎
무적%하충성%종왜%로석기%우도%진기규
结肠癌%小干扰RNA%Wip1基因%化疗敏感性
結腸癌%小榦擾RNA%Wip1基因%化療敏感性
결장암%소간우RNA%Wip1기인%화료민감성
Colon cancer%Small interfering RNA%Wip1 gene%Chemotherapy sensitivity
目的：观察靶向Wip1基因的特异性siRNA序列对结肠癌细胞Wip1基因的抑制效应，探讨Wip1基因沉默对结肠癌细胞化疗敏感性的影响。方法：将Wip1-811 siRNA转染入Wip1高表达的RKO结肠癌细胞株，采用real-time PCR法检测Wip1 mRNA的表达，采用Western blotting方法检测Wip1蛋白表达，采用MTS方法检测结肠癌细胞活力，流式细胞术检测细胞凋亡及细胞周期。结果： Wip1-811 siRNA明显抑制Wip1的表达。抑制Wip1基因表达后，转染组RKO结肠癌细胞对抗肿瘤药物的敏感性增加，5-氟尿嘧啶处理组RKO结肠癌细胞活力由（89.4±6.6）％降至（74.7±3.9）％（P＜0.05），奥沙利铂处理组细胞活力由（77.9±2.4）％降至（66.7±2.9）％（P＜0.05）。转染Wip1-811 siRNA 后，5-氟尿嘧啶处理组细胞凋亡比例由（7.7±0.5）％升至（12.3±3.2）％（P＜0.05）；奥沙利铂处理组细胞凋亡比例由（14.7±2.1）％升至（34.0±2.1）％（P＜0.05）。结论：沉默Wip1基因表达能增强结肠癌细胞的化疗敏感性。
目的：觀察靶嚮Wip1基因的特異性siRNA序列對結腸癌細胞Wip1基因的抑製效應，探討Wip1基因沉默對結腸癌細胞化療敏感性的影響。方法：將Wip1-811 siRNA轉染入Wip1高錶達的RKO結腸癌細胞株，採用real-time PCR法檢測Wip1 mRNA的錶達，採用Western blotting方法檢測Wip1蛋白錶達，採用MTS方法檢測結腸癌細胞活力，流式細胞術檢測細胞凋亡及細胞週期。結果： Wip1-811 siRNA明顯抑製Wip1的錶達。抑製Wip1基因錶達後，轉染組RKO結腸癌細胞對抗腫瘤藥物的敏感性增加，5-氟尿嘧啶處理組RKO結腸癌細胞活力由（89.4±6.6）％降至（74.7±3.9）％（P＜0.05），奧沙利鉑處理組細胞活力由（77.9±2.4）％降至（66.7±2.9）％（P＜0.05）。轉染Wip1-811 siRNA 後，5-氟尿嘧啶處理組細胞凋亡比例由（7.7±0.5）％升至（12.3±3.2）％（P＜0.05）；奧沙利鉑處理組細胞凋亡比例由（14.7±2.1）％升至（34.0±2.1）％（P＜0.05）。結論：沉默Wip1基因錶達能增彊結腸癌細胞的化療敏感性。
목적：관찰파향Wip1기인적특이성siRNA서렬대결장암세포Wip1기인적억제효응，탐토Wip1기인침묵대결장암세포화료민감성적영향。방법：장Wip1-811 siRNA전염입Wip1고표체적RKO결장암세포주，채용real-time PCR법검측Wip1 mRNA적표체，채용Western blotting방법검측Wip1단백표체，채용MTS방법검측결장암세포활력，류식세포술검측세포조망급세포주기。결과： Wip1-811 siRNA명현억제Wip1적표체。억제Wip1기인표체후，전염조RKO결장암세포대항종류약물적민감성증가，5-불뇨밀정처리조RKO결장암세포활력유（89.4±6.6）％강지（74.7±3.9）％（P＜0.05），오사리박처리조세포활력유（77.9±2.4）％강지（66.7±2.9）％（P＜0.05）。전염Wip1-811 siRNA 후，5-불뇨밀정처리조세포조망비례유（7.7±0.5）％승지（12.3±3.2）％（P＜0.05）；오사리박처리조세포조망비례유（14.7±2.1）％승지（34.0±2.1）％（P＜0.05）。결론：침묵Wip1기인표체능증강결장암세포적화료민감성。
AIM:To observe the inhibitory effect of siRNA targeting to Wip1 gene on the Wip1 gene expression in the colon cancer cells and to investigate the influence of Wip1 gene silencing on the chemotherapy sensitivity of colon cancer cells.METHODS:Wip1-811 siRNA targeting to Wip1 gene was transfected into RKO colon cancer cells with high expression of Wip1 gene.The mRNA expression of Wip1 was measured by real-time PCR.The protein level of Wip1 was detected by Western blotting.The viability of RKO colon cancer cells was measured by MTS assay.The cell apoptosis and cell cycle were analyzed by flow cytometry.RESULTS: Wip1-811 siRNA efficiently inhibited the expression of Wip1 at mRNA and protein levels.The enhanced chemotherapy sensitivity of RKO colon cancer cells was observed after inhibition of Wip1 gene expression.The viability of RKO colon cancer cells was decreased from (89.4 ±6.6)%to (74.7 ±3.9)%af-ter treated with 5-fluorouracil (P<0.05) and decreased from (77.9 ±2.4)%to (66.7 ±2.9)%after treated with oxali-platin ( P<0.05 ) .The cell apoptotic rate was increased from ( 7.7 ±0.5 )% to ( 12.3 ±3.2 )% and from ( 14.7 ± 2.1)% to (34.0 ±2.1)% when RKO colon cancer cells were treated with 5-fluorouracil and oxaliplatin, respectively (P<0.05).CONCLUSION:Wip1 gene silencing enhances chemotherapy sensitivity of colon cancer cells.
