c—myc反义RNA转录体的构建及其对HL60细胞恶性表型的影响

Construction of c-myc RNA Antisense Expression Plasmid and Its Effects on the Malignant Phenotype of HL60 Cell Line

本研究构建了c-myc反义RNA真核细胞可诱导表达质粒PGXC.用PGXC及PSV_(2)neo质粒共转染人早幼粒细胞白血病细胞HL60,经G418抗性筛选得到转染细胞HL_(60)L^R,与亲本细胞相比其生长速率明显减低;细胞周期中G1期细胞增多,而S期细胞相应减少;细胞形态及功能呈现分化诱导;在软琼脂上不能形成集落;该细胞在裸鼠体内的致瘤性亦显著减弱以至消失.

The inducible c-myc antisense RNA expression plasmid PGXC expressed in mammalian cells was constructed. PGXC was cotransferred with plasmid pSV2neo which confers resistance to Geneticin (G418) to human promyclocytic leukemia cell line HL60. Following the promotion of c-myc antisense RNA expression with Cd^(2+) the growth rate of transfectant HL_(60)~R-9 was significantly slower than that of the parent cells HL60. Flow cytometric analysis indicated a marked increase in Gl fraction and decrease in S-phase population in the cell cycle of transfectants. The differentiation of cellular morphology and function of HL_(60)~R-9 was induced. HL_(60)~R-9 did not form colonies on soft agar and had reduced or lost the tumorigenicity in nude mice.