摘要
目的应用RNA干扰技术沉默Pinl表达后，观察其对乳腺癌细胞增殖、侵袭和凋亡的影响。方法采用脂质体介导法将Pinl干扰片段PinlsiRNA和对照片段controlsiRNA转染入乳腺癌细胞系MDA—MB一23l后，利用RT—PCR和Westernblot法检测Pinl基因在mRNA和蛋白水平的表达情况，并应用MTT法、Transwell法和AnnexinV—FITC／PI试剂盒及流式细胞仪技术分析检测Pinl对MDA—MB一23l细胞增殖、侵袭和凋亡的影响。结果Pinl在乳腺癌细胞系中高表达。与未处理组及转染对照片段controlsiRNA组相比，沉默Pinl表达后，PinlmRNA（P〈0．05）和蛋白（P〈0．05）表达均明显下降，细胞生长增殖能力[P〉0．05（第1天），P〈0．01（第2—4天）]减弱，细胞侵袭数目（6．61±0．53，P〈0．05）减少，细胞凋亡率（31．5％±3．06％，P〈0．05）显著增加。结论沉默Pinl表达后可抑制乳腺癌细胞增殖和侵袭，促进乳腺癌细胞凋亡。
Objective To investigate the effects of Pinl reduction on cell proliferation, invasion and apoptosis of breast cancer MDA - MB - 231 cells by silencing Pinl gene using RNA interference technique. Methods MDA - MB - 231 cells were transfected with Pinl siRNA and control siRNA using Lipofectamine 2000. The expression of SIAH1 mRNA and protein were detected by RT - PCR and West- ern blot. The cell proliferation was analyzed by MTT and the cell invasion was tested by Transwell. Annexin V - FITC/PI reagent kit and Flow cytometry assay were used to analyze cell apoptosis. Results The expression of Pinl was higher in breast cancer ceils than in nor- mal mammary epithelial cells. Compared with untreated and the cells transfected with control siRNA, the level of Pinl mRNA and protein（P 〈 0.05） were decreased in the cells transfected with the Pinl siRNA. The cell growth rate [ P 〉 0.05 （ day 1 （day 2-4） ] was slower, the numbers of cell invasion （6.61 ~ 0.53 ,P 〈 0.05 ）were decreased, and the apoptotic rate 3.06% ,P 〈 0.05） was increased in MDA - MB - 231 cells transfected with Pinl siRNA. Conclusion Konck down of Pin siRNA may suppress cell proliferation and invasion, promote cell apoptosis in breast cancer ceils. P 〈0.05） ,P〈0.01 31.5% ＋ with Pinl
出处
《医学研究杂志》
2013年第12期36-39,共4页
Journal of Medical Research
基金
国家自然科学基金(青年科学基金项目)资助项目(81201853)
浙江省自然科学基金资助项目(Y2110620
Y2111209)
关键词
乳腺癌
PIN1
增殖
侵袭
凋亡
Breast cancer
Pinl
Proliferation
Invasion
Apoptosis
