摘要
目的 探讨脂质体介导c myc反义核酸的抗肿瘤作用。方法 用人工合成互补于c myc基因 5′端转录起始部位的反义 寡核苷酸 (AS ODN) ,直接转染HL 6 0细胞 ,或以脂质体lipofectin为载体进行基因转染实验 ,研究HL 6 0细胞在基因治疗后的形态和功能变化。结果 脂质体介导的c myc反义核酸可抑制约 5 0 %HL 6 0细胞的增殖 ;促进细胞分化成熟 ,四唑氮蓝 (NBT)还原率从空白组 2 9%上升至 5 6 % ,并诱导 17%的细胞凋亡。免疫组化显示c myc蛋白阳性率从空白组 ( 80 0 0± 3 0 2 ) %下降至 ( 41 6 7± 4 5 6 ) % ;逆转录 聚合酶链反应 (RT PCR)显示c myc基因的PCR扩增条带明显减弱 ;DNA电泳显示典型的凋亡梯形降解条带。单纯ODN无上述作用。结论 脂质体介导AS ODN可抑制HL 6 0细胞的恶性表型。应用脂质体可提高细胞对反义核酸的摄取率 ,减少反义核酸用量。
Objective To investigate a delivery system for administering antisense oligodeoxynucleotide (AS-ODN) to HL-60 cells and study the antitumor effects of c-myc antisense oligodeoxynucleotide. Methods Human leukemia cell line HL-60 was transfected with a synthetic oligodeoxynucleotide(ODN) complementary to the 5′ end region of c-myc gene. In addition, the changes of function and morphology were observed after transfection of cationic liposome. Results HL-60 cells incubated with c-myc AS-ODN/ lipofectin complex were about 50% decreased in the 4th day as compared with control cells (P<005). About 56% of HL-60 cells exposed to c-myc AS-ODN/lipofectin complex were NBT-positive and showed morphological changes with differentiated phenotype compared with 29% of untreated cells. The expression of the c-myc in the treated cells decreased markedly. The levels of c-myc protein after treatment decreased from (8000±3 02)% to (4167±456)% (P<001). c-myc AS-ODN induced HL-60 cells apoptosis which morphological changes of apoptotic cells were investigated by fluorescence microsopy. A ladder-like pattern of DNA fragmentation was demonstrated on agorose gel electrophoresis and 17% apoptotic cells were found with flow cytometry. Conclusions Antisense oligodeoxynucleotide can suppress the maligant phenotype of HL-60 cells and enhancement of the effect was observed by adding lipofectin.
出处
《中华内科杂志》
CAS
CSCD
北大核心
2000年第10期660-663,I023,共5页
Chinese Journal of Internal Medicine
