中华胰腺病杂志
中華胰腺病雜誌
중화이선병잡지
CHINESE JOURNAL OF PANCREATOLOGY
2009年
1期
27-30
,共4页
张海峰%周国雄%丁晓凌%周德霞
張海峰%週國雄%丁曉凌%週德霞
장해봉%주국웅%정효릉%주덕하
胰腺肿瘤%RNA干扰%5-脂氧合酶%细胞增殖%细胞凋亡
胰腺腫瘤%RNA榦擾%5-脂氧閤酶%細胞增殖%細胞凋亡
이선종류%RNA간우%5-지양합매%세포증식%세포조망
Pancreatic neoplasms%RNA interference%5-lipoxygenase%Proliferation inhibition%Apoptosis
目的 应用RNA干扰(RNAi)技术阻断5-脂氧合酶(5-LOX)的表达,观察其对人胰腺癌细胞SW1990增殖及凋亡的作用.方法 构建4个靶向5-LOX的小干扰RNA(small interference,siRNA)和1个阴性对照siRNA质粒表达载体,采用Lipofectamine TM2000法转染SWl990细胞,RT-PCR和Western blot检测RNAi后SWl990细胞的5-LOX mRNA和蛋白表达,MTT法检测细胞的增殖抑制率,流式细胞仪检测细胞凋亡率.结果 阴性对照和4个序列特异性的siRNA对SWl990细胞5-LOXmRNA表达的抑制率分别为(3.0±1.4)%、(18.8±1.5)%、(53.5±2.3)%、(56.1±2.0)%、(52.4±2.5)%;5-LOX蛋白表达抑制率分别为(4.5±2.0)%、(18.1±2.5)%、(50.4±4.3)%、(48.9±4.4)%、(45.9±4.0)%.转染24 h后癌细胞增殖抑制率分别为(2.1±1.0)%、(5.5±1.3)%、(11.9±1.2)%、(13.4±1.1)%、(13.8±1.3)%.;48 h的增殖抑制率分别为(3.0±1.3)%、(16.0±2.2)%、(25.7±2.5)%、(25.3±3.1)%、(27.2±3.2)%.转染24 h细胞凋亡率分别为(2.0±0.8)%、(5.3±1.0)%、(10.6±1.2)%、(12.4±1.0)%、(10.6±0.9)%;转染48 h的凋亡率分别为(3.0±1.0)%、(7.1±1.1)%、(17.5±0.9)%、(21.5±1.1)%、(15.7±1.0)%.结论 通过RNAi可以有效、特异地阻断SWl990细胞5-LOX的表达,并可以有效抑制肿瘤细胞的增殖,促进肿瘤细胞凋亡.
目的 應用RNA榦擾(RNAi)技術阻斷5-脂氧閤酶(5-LOX)的錶達,觀察其對人胰腺癌細胞SW1990增殖及凋亡的作用.方法 構建4箇靶嚮5-LOX的小榦擾RNA(small interference,siRNA)和1箇陰性對照siRNA質粒錶達載體,採用Lipofectamine TM2000法轉染SWl990細胞,RT-PCR和Western blot檢測RNAi後SWl990細胞的5-LOX mRNA和蛋白錶達,MTT法檢測細胞的增殖抑製率,流式細胞儀檢測細胞凋亡率.結果 陰性對照和4箇序列特異性的siRNA對SWl990細胞5-LOXmRNA錶達的抑製率分彆為(3.0±1.4)%、(18.8±1.5)%、(53.5±2.3)%、(56.1±2.0)%、(52.4±2.5)%;5-LOX蛋白錶達抑製率分彆為(4.5±2.0)%、(18.1±2.5)%、(50.4±4.3)%、(48.9±4.4)%、(45.9±4.0)%.轉染24 h後癌細胞增殖抑製率分彆為(2.1±1.0)%、(5.5±1.3)%、(11.9±1.2)%、(13.4±1.1)%、(13.8±1.3)%.;48 h的增殖抑製率分彆為(3.0±1.3)%、(16.0±2.2)%、(25.7±2.5)%、(25.3±3.1)%、(27.2±3.2)%.轉染24 h細胞凋亡率分彆為(2.0±0.8)%、(5.3±1.0)%、(10.6±1.2)%、(12.4±1.0)%、(10.6±0.9)%;轉染48 h的凋亡率分彆為(3.0±1.0)%、(7.1±1.1)%、(17.5±0.9)%、(21.5±1.1)%、(15.7±1.0)%.結論 通過RNAi可以有效、特異地阻斷SWl990細胞5-LOX的錶達,併可以有效抑製腫瘤細胞的增殖,促進腫瘤細胞凋亡.
목적 응용RNA간우(RNAi)기술조단5-지양합매(5-LOX)적표체,관찰기대인이선암세포SW1990증식급조망적작용.방법 구건4개파향5-LOX적소간우RNA(small interference,siRNA)화1개음성대조siRNA질립표체재체,채용Lipofectamine TM2000법전염SWl990세포,RT-PCR화Western blot검측RNAi후SWl990세포적5-LOX mRNA화단백표체,MTT법검측세포적증식억제솔,류식세포의검측세포조망솔.결과 음성대조화4개서렬특이성적siRNA대SWl990세포5-LOXmRNA표체적억제솔분별위(3.0±1.4)%、(18.8±1.5)%、(53.5±2.3)%、(56.1±2.0)%、(52.4±2.5)%;5-LOX단백표체억제솔분별위(4.5±2.0)%、(18.1±2.5)%、(50.4±4.3)%、(48.9±4.4)%、(45.9±4.0)%.전염24 h후암세포증식억제솔분별위(2.1±1.0)%、(5.5±1.3)%、(11.9±1.2)%、(13.4±1.1)%、(13.8±1.3)%.;48 h적증식억제솔분별위(3.0±1.3)%、(16.0±2.2)%、(25.7±2.5)%、(25.3±3.1)%、(27.2±3.2)%.전염24 h세포조망솔분별위(2.0±0.8)%、(5.3±1.0)%、(10.6±1.2)%、(12.4±1.0)%、(10.6±0.9)%;전염48 h적조망솔분별위(3.0±1.0)%、(7.1±1.1)%、(17.5±0.9)%、(21.5±1.1)%、(15.7±1.0)%.결론 통과RNAi가이유효、특이지조단SWl990세포5-LOX적표체,병가이유효억제종류세포적증식,촉진종류세포조망.
Objective To investigate the effects of inhibition of 5-LOX by RNA interference on proliferation suppression and apoptosis induction of pancreatic cancer cell line. Methods Plasmid expression vectors containing four 5-LOX siRNA array and one negative control array were established, resoectively, and were transfected into pancreatic cancer cell line SW1990 with Lipofectamine TM2000. Cell proliferation inhibition rate was measured by MTT assay; apoptotic rate was examined by flow cytometry. Results The inhibitory rate of expression of 5-LOX mRNA in negative control group and four 5-LOX siRNA groups was (3.0 ±1.4)%, (18.8±1.5)%, (53.5±2.3)%, (56.1±2.0)%, (52.4±2.5)%; the inhibitory rate of expression of 5-LOX protein was (4.5 ± 2.0) %, (18.1 ± 2.5) %, (50.4 ± 4.3) %, (48.9 ± 4.4) %, (45.9 ± 4.0) %. The inhibitory rates of cancer cell proliferation at 24 h and 48 h after the transfection were (2.1±1.0)%, (5.5±1.3)%, (11.9±1.2)%, (13.4±1.1)%, (13.8±1.3)% and (3.0±1.3)%, (16.0 ± 2.2) %, (25.7 ± 2.5) %, (25.3 ± 3.1) %, (27.2 ± 3.2) %, respectively. The apoptotic rates at 48 h after the transfection were (3.0 ± 1.0) %, (7.1 ± 1.10%, (17.5 ± 0. 9) %, (21.5 ± 1.1) %, (15.7 ± 1. 0)%, respectively. Conclusions The plasmid vector containing siRNA against 5-LOX could suppress 5-LOX expression in SW1990 cells effectively and specifically, and could inhibit proliferation and induce the apoptosis of pancreatic cancer cells.
