中华放射医学与防护杂志
中華放射醫學與防護雜誌
중화방사의학여방호잡지
Chinese Journal of Radiological Medicine and Protection
2010年
4期
410-413
,共4页
张克君%唐立岷%焦学龙%张炳远%孙传东%卢云%曹红诗
張剋君%唐立岷%焦學龍%張炳遠%孫傳東%盧雲%曹紅詩
장극군%당립민%초학룡%장병원%손전동%로운%조홍시
Slug基因%干扰%PUMA基因%放射敏感性
Slug基因%榦擾%PUMA基因%放射敏感性
Slug기인%간우%PUMA기인%방사민감성
Slug gene%Short interferencing RNA%PUMA gene%Radiosensitivity
目的 探讨siRNA干扰Slug基因表达对胰腺癌AsPC-1细胞γ射线敏感性的影响.方法 以0、10、50、100感染复数(MOI)的rAAV-EGFP-Slug-siRNA(Slug-siRNA)转染AsPC-1细胞72 h,免疫组织化学法和Western blotting检测转染前后细胞内Slug和PUMA表达.采用4 Gy γ射线细胞进行照射,MTT比色法和Hoechst 33342和PI双染法检测对照组、转染组(MOI 50)、照射组、转染(MOI 50)+照射组细胞存活率和凋亡率,DNA琼脂糖凝胶电泳检测细胞凋亡,Giemsa染色观察细胞形态.结果 细胞受到MOI为10、50、100作用72 h后,Slug蛋白表达相对值分别为0.831±0.14、0.546±0.12和0.178±0.08,明显低于对照组1.17±0.152,差异有统计学意义(F=4.992,P＜0.05);而细胞受到MOI为10、50、100作用72 h,PUMA蛋白表达相对值分别为0.325±0.07、0.593±0.11和0.978±0.12,明显高于对照组0.143±0.04,差异有统计学意义(F=4.324,P＜0.05);转染(MOI 50)+照射组细胞增殖抑制率为(78.76±9.36)%,明显高于转染组和单独射线照射组[(43.68±6.71)%和(19.25±3.72)%],差异有统计学意义(F=5.056,P＜0.05).另外,转染(MOI 50)+照射组细胞的凋亡现象较其他组更加明显.结论 siRNA干扰Slug基因表达能够解除Slug对PUMA的抑制,增强胰腺癌细胞对γ射线的敏感性.
目的 探討siRNA榦擾Slug基因錶達對胰腺癌AsPC-1細胞γ射線敏感性的影響.方法 以0、10、50、100感染複數(MOI)的rAAV-EGFP-Slug-siRNA(Slug-siRNA)轉染AsPC-1細胞72 h,免疫組織化學法和Western blotting檢測轉染前後細胞內Slug和PUMA錶達.採用4 Gy γ射線細胞進行照射,MTT比色法和Hoechst 33342和PI雙染法檢測對照組、轉染組(MOI 50)、照射組、轉染(MOI 50)+照射組細胞存活率和凋亡率,DNA瓊脂糖凝膠電泳檢測細胞凋亡,Giemsa染色觀察細胞形態.結果 細胞受到MOI為10、50、100作用72 h後,Slug蛋白錶達相對值分彆為0.831±0.14、0.546±0.12和0.178±0.08,明顯低于對照組1.17±0.152,差異有統計學意義(F=4.992,P＜0.05);而細胞受到MOI為10、50、100作用72 h,PUMA蛋白錶達相對值分彆為0.325±0.07、0.593±0.11和0.978±0.12,明顯高于對照組0.143±0.04,差異有統計學意義(F=4.324,P＜0.05);轉染(MOI 50)+照射組細胞增殖抑製率為(78.76±9.36)%,明顯高于轉染組和單獨射線照射組[(43.68±6.71)%和(19.25±3.72)%],差異有統計學意義(F=5.056,P＜0.05).另外,轉染(MOI 50)+照射組細胞的凋亡現象較其他組更加明顯.結論 siRNA榦擾Slug基因錶達能夠解除Slug對PUMA的抑製,增彊胰腺癌細胞對γ射線的敏感性.
목적 탐토siRNA간우Slug기인표체대이선암AsPC-1세포γ사선민감성적영향.방법 이0、10、50、100감염복수(MOI)적rAAV-EGFP-Slug-siRNA(Slug-siRNA)전염AsPC-1세포72 h,면역조직화학법화Western blotting검측전염전후세포내Slug화PUMA표체.채용4 Gy γ사선세포진행조사,MTT비색법화Hoechst 33342화PI쌍염법검측대조조、전염조(MOI 50)、조사조、전염(MOI 50)+조사조세포존활솔화조망솔,DNA경지당응효전영검측세포조망,Giemsa염색관찰세포형태.결과 세포수도MOI위10、50、100작용72 h후,Slug단백표체상대치분별위0.831±0.14、0.546±0.12화0.178±0.08,명현저우대조조1.17±0.152,차이유통계학의의(F=4.992,P＜0.05);이세포수도MOI위10、50、100작용72 h,PUMA단백표체상대치분별위0.325±0.07、0.593±0.11화0.978±0.12,명현고우대조조0.143±0.04,차이유통계학의의(F=4.324,P＜0.05);전염(MOI 50)+조사조세포증식억제솔위(78.76±9.36)%,명현고우전염조화단독사선조사조[(43.68±6.71)%화(19.25±3.72)%],차이유통계학의의(F=5.056,P＜0.05).령외,전염(MOI 50)+조사조세포적조망현상교기타조경가명현.결론 siRNA간우Slug기인표체능구해제Slug대PUMA적억제,증강이선암세포대γ사선적민감성.
Objective To explore the influence of PUMA on radiosensitivity of pancreatic cancer AsPC-1 cells after Slug gene inhibition by transfected short interferencing RNA(siRNA). Methods The AsPC-1 cells were infected with MOI 10,50,100 for 72 h, respectively. The expression of Slug and PUMA was analyzed by Western blotting and immunohistochemistry methods. The transfected and control cells were exposed to 4 Gy γ-rays. The cells inhibition rate was examined by MTT, Hoechst 33342 and IP double staining. DNA ladder and Giemsa staning was used to observe apoptosis. Results The relative value of Slug expression was 0.831 ±0.14,0. 546 ±0.12 and 0.178 ±0.08 after AsPC-1 was infected with Slug-siRNA ( MOI 10,50,100) for 72 h, significantly lower than that of control group ( F = 4. 992,P ＜ 0.05 ).The relative value of PUMA was 0. 325 ±0. 07,0. 593 ±0. 11 and 0. 978 ±0. 12, after AsPC-1 was infected with Slug-siRNA ( MOI 10,50,100) for 72 h, significantly higher than that of control group ( F = 4. 324,P ＜ 0. 05 ). The cell proliferation rate was ( 78.76 ± 9. 36 ) % in transfection combined with radiosensitivity group, significantly higher than that of transfection group [ ( 43.68 ± 6.71 ) % ] and radiosensitivity group alone [( 19.25 ± 3.72)% ] (F = 5.056, P ＜ 0.05). The apoptosis of transfection combined with radiosensitivity group was significantly higher than that of others. Conclusions Slug gene targeting siRNA could inhibit the expression of Slug, and consequently increase the activation of PUMA expression, and so enhance the radiosensitivity to γ-rays.
