中华胰腺病杂志
中華胰腺病雜誌
중화이선병잡지
CHINESE JOURNAL OF PANCREATOLOGY
2013年
4期
252-255
,共4页
胰腺肿瘤%ABCC4%细胞增殖%RNA干扰%慢病毒感染
胰腺腫瘤%ABCC4%細胞增殖%RNA榦擾%慢病毒感染
이선종류%ABCC4%세포증식%RNA간우%만병독감염
Pancreatic neoplasms%ABCCA%Cell,proliferation%RNA interference%Lentivirus infections
目的 检测ABCC4基因沉默后对人胰腺癌PANC1、BxPC-3细胞增殖及细胞周期的影响.方法 构建插入靶向ABCC4的shRNA片段的重组慢病毒载体,感染人胰腺癌细胞株PANC1和BxPC-3.采用实时定量PCR法和蛋白质印迹法检测感染细胞的ABCC4 mRNA及蛋白表达,克隆形成实验测定感染细胞形成的克隆数,流式细胞仪检测感染细胞的细胞周期.结果 成功构建了插入靶向ABCC4的shRNA片段的重组慢病毒载体.重组慢病毒感染PANC1及BxPC-3细胞后,细胞ABCC4 mRNA表达被抑制(0.28 0.01比1.00±0.03,0.22 ±0.02比1.00 ±0.03,P值均＜0.05)；ABCC4蛋白表达亦显著下调；感染的PANC1细胞克隆形成数量显著减少(4比65,P＜0.05)；细胞周期阻滞在G1期[(54.98±1.78)％比(42.93±0.88)％,(68.55±0.75)％比(54.76 ±0.29)％].结论 沉默人胰腺癌PANC1和BxPC-3细胞的ABCC4基因表达可显著抑制癌细胞的增殖,使细胞阻滞于G1期.
目的 檢測ABCC4基因沉默後對人胰腺癌PANC1、BxPC-3細胞增殖及細胞週期的影響.方法 構建插入靶嚮ABCC4的shRNA片段的重組慢病毒載體,感染人胰腺癌細胞株PANC1和BxPC-3.採用實時定量PCR法和蛋白質印跡法檢測感染細胞的ABCC4 mRNA及蛋白錶達,剋隆形成實驗測定感染細胞形成的剋隆數,流式細胞儀檢測感染細胞的細胞週期.結果 成功構建瞭插入靶嚮ABCC4的shRNA片段的重組慢病毒載體.重組慢病毒感染PANC1及BxPC-3細胞後,細胞ABCC4 mRNA錶達被抑製(0.28 0.01比1.00±0.03,0.22 ±0.02比1.00 ±0.03,P值均＜0.05)；ABCC4蛋白錶達亦顯著下調；感染的PANC1細胞剋隆形成數量顯著減少(4比65,P＜0.05)；細胞週期阻滯在G1期[(54.98±1.78)％比(42.93±0.88)％,(68.55±0.75)％比(54.76 ±0.29)％].結論 沉默人胰腺癌PANC1和BxPC-3細胞的ABCC4基因錶達可顯著抑製癌細胞的增殖,使細胞阻滯于G1期.
목적 검측ABCC4기인침묵후대인이선암PANC1、BxPC-3세포증식급세포주기적영향.방법 구건삽입파향ABCC4적shRNA편단적중조만병독재체,감염인이선암세포주PANC1화BxPC-3.채용실시정량PCR법화단백질인적법검측감염세포적ABCC4 mRNA급단백표체,극륭형성실험측정감염세포형성적극륭수,류식세포의검측감염세포적세포주기.결과 성공구건료삽입파향ABCC4적shRNA편단적중조만병독재체.중조만병독감염PANC1급BxPC-3세포후,세포ABCC4 mRNA표체피억제(0.28 0.01비1.00±0.03,0.22 ±0.02비1.00 ±0.03,P치균＜0.05)；ABCC4단백표체역현저하조；감염적PANC1세포극륭형성수량현저감소(4비65,P＜0.05)；세포주기조체재G1기[(54.98±1.78)％비(42.93±0.88)％,(68.55±0.75)％비(54.76 ±0.29)％].결론 침묵인이선암PANC1화BxPC-3세포적ABCC4기인표체가현저억제암세포적증식,사세포조체우G1기.
Objective To determine the effect of ABCC4 gene silencing on cell proliferation and cell cycle in human pancreatic cancer cell lines PANC1 and BxPC-3.Methods PANC1 and BxPC-3 pancreatic cancer cells were transfected with a lentivirus expressing an ABCCA short hairpin RNA (shRNA).ABCC4 mRNA and protein expression of transfected cells was determined by RT-PCR and Western blot,colony formation ability was measured by colony assay,and cell cycle change was investigated by the flow cytometric analysis.Results Lentivirus expressing an ABCC4 short hairpin RNA (shRNA) was successfully established.After transfection with shRNA lentivirus,ABCC4 mRNA and protein expression were significantly inhibited (0.28 ±0.01 vs 1.00 ±0.03,0.22 ±0.02 vs 1.00 ±0.03,P ＜0.05).Colony formation ability was significantly decreased (4 vs 65,P ＜0.05),and cell cycle was significantly blocked at G1 phase [(54.98 ±1.78) ％ vs (42.93 ± 0.88) ％,(68.55 ± 0.75) ％ vs (54.76 ± 0.29) ％].Conclusions ABCC4 gene silencing can significantly inhibit cell proliferation of human pancreatic cancer cell lines PANC1 and BxPC-3,and block the cells at G1 phase.
