摘要
目的观察乳腺癌细胞MDA—MB-231 osteopontin（OPN）RNA靶向干扰效果。方法采用OPNmRNA干扰质粒转染MDA-MB-231细胞，建立OPN基因沉默细胞株，采用RT-PCR、蛋白质印迹法从mRNA和蛋白水平检测乳腺癌细胞OPN表达量的变化，侵袭试验检测OPN下调对细胞侵袭能力的影响，MTT法测细胞增殖能力的改变，裸鼠成瘤后观察OPNRNA干扰对肿瘤细胞生长的影响及对移植瘤乏氧程度的影响。结果与MDA—MB-231细胞相比，OPN沉默细胞在常氧和乏氧状态下OPN表达均显著下调（均P〈0．05），OPN稳定沉默细胞侵袭力和增殖能力下降（均P〈0．01），OPNRNAi能明显抑制裸鼠移植瘤的生长（P〈0．05），且移植瘤内乏氧程度降低（P〈0．05）。结论OPNRNA干扰可抑制乳腺癌细胞MDA—MB-231细胞侵袭力和增殖能力，降低移植瘤的乏氧程度。
Objective To investigate the role of OPN in human breast cancer cell line （ MDA-MB- 231 ） by using small interfering RNA to specifically knockdown OPN expression. Methods OPN ShRNA expression vector was stably transfected to MDA-MB-231 cell line. The expression of OPN mRNA and protein were analyzed using reverse transcription polymerase chain reaction （RT-PCR） and Western blot, respectively. The growth of MDA-MB-231 cells were observed by MTF. The effect of OPN siRNA on the transplanted tumor growth and tumor hypoxia were assessed in nude mice. Results The expression level of OPN in MDA-MB-231 cells were significantly lower under hypoxia or normoxia（ P 〈 0. 05 ）. OPN silence with RNAi significantly inhibited the invasion ability and proliferation of MDA-MB-231 cell lines （ P 〈 0. 01 ）. Inhibition of OPN with RNAi significantly inhibited the growth ability of MDA-MB-231 cells in vivo（P 〈 0. 05）. The tumor hypoxia significantly decreased（P 〈 0.05）. Conclusions OPN silence with RNAi can effectively inhibit cell proliferation and tumor growth of MDA-MB-231 cells, and decrease the bypoxia level of MDA-MB-231 transplanted tumor in nude mice.
出处
《中华普通外科杂志》
CSCD
北大核心
2012年第4期322-325,共4页
Chinese Journal of General Surgery
基金
山东省自然基金项目（Y2008C167）
