Rb反义寡核苷酸对人结肠癌细胞凋亡及化疗药敏的影响

目的　观察Rb反义寡核苷酸 (AS ODN)对人结肠癌(HT 2 9)细胞凋亡及化疗药物敏感性的影响。方法　人工合成RbAS ODN经脂质体 (Lip)包裹后作用HT 2 9细胞。采用免疫组化、WesternBlot检测转染前后Rb蛋白表达状况 ,流式细胞术测定HT 2 9细胞转染前后 5 Fu诱导凋亡程度 ,MTT法观察化疗药敏的变化。结果　经RbAS ODN处理的HT 2 9细胞Rb蛋白表达量明显下降 ,RbAS ODN处理的HT 2 9细胞加 5 Fu作用后与对照组比较 ,细胞凋亡率明显下降 ,化疗药物敏感性降低。结论　RbAS ODN阳离子脂质体转染具有抑制化疗药诱导大肠癌HT 2 9细胞凋亡的作用及降低化疗药物敏感性作用。

The effects of Rb AS-ODN on apoptosis and drug sensitivity of colon cancer cells

AIM To investigate the effects of Rb AS-ODN on apoptosis and drug sensitivity of colon cancer cells. METHODS A cationic liposome was selected as a delivery vehicle. HT-29 cells were transfected with Rb AS-ODN,S-ODN and LP in vitro. The expression of pRb treated with or without AS-ODN,S-ODN and LP in HT-29 cells were observed by Streptavidin-Peroxidase(S-P) immunohistochemical method and Western Blot analysis. The Flow cytometer(FCM) was used to examine apoptosis induced by 5-Fu in HT-29 cells. The MTT assay was used to measure chemotherapeutic agent-sensitivityFQ(9。46,X-WZ]- of HT-29 cells treated with 5-Fu. RESULTS The level of pRb expression in the HT-29 colon cancer line transfected with Rb AS-ODN obviously reduced compared with that of controls ,which were treated with S-ODN,LP and non-transfected(P<0.05). The percentage of the apoptotic cells and drug-sensitivity of HT-29 cells of the treatment with Rb AS-ODN to 5-Fu decreased compared with that of the control(P<0.05). CONCLUSION The transfection of Rb AS-ODN delivered by cationic liposome could inhibit apoptosis induced by 5-Fu in HT-29 cells and decrease drug-sensitivity of HT-29 cells to 5-Fu.-