Apollon反义寡核苷酸对K562细胞增殖、凋亡和耐药的影响

Effects of Apollon antisense oligonucleotide on proliferation and apoptosis of K562 cells

目的探讨Apollon反义寡核苷酸（antisense oligonucleotide，ASODN）对白血病细胞K562增殖、凋亡及耐药的影响。方法设计合成特异性的Apollon硫代磷酸ASODN及其对照错义寡核苷酸（missenseoli．gonucleotide，MSODN），脂质体介导转染K．562细胞后，采用四甲基偶氮唑蓝（MTF）比色法检测K562细胞增殖，AnnexinV—FITC法检测细胞的凋亡，并用MTI＂法检测K562细胞对依托泊苷、长春新碱及联合用ASODN的耐药性。结果ApollonASODN对K562细胞的生长抑制作用随浓度和时间增加而增加。ApollonASODN在终浓度为600nmol/L作用K562细胞时，能明显抑制其增殖，K562细胞凋亡率显著增加（P〈0．01），联合用药组对细胞的半数抑制浓度低于单药组。结论ApollonASODN可下调Apollon基因表达，有效抑制K562细胞的增殖，并促进K562细胞的凋亡，ApollonASODN与依托泊苷和长春新碱联合作用可降低细胞生存率，降低细胞耐药性。

Objective To investigate the effects of Apollon antisense oligonucleotide （ASODN） on prolifera- tion, apoptosis and drug resistance of human leukemia（K562） cells. Methods Specific phosphorothioate ASODN and missense oligonueleotide （MSODN） of Apollon mRNA were synthesized and transfeeted into K562 cells following cationic liposome. The proliferation inhibition of K562 cells was assessed by MTT. The apoptosis rate was detected by Annexin V-FITC. The sensitivity of K562 cells to etoposide and vineristine was detected by MTF. Results Apollon an- tisense oligonucleotide inhibition of K562 cells with the concentration and time increased. ASODN at a final concentra- tion of 600 nmol/L could significantly inhibit the K562 cell proliferation. The apoptosis rate was apparently increased （P 〈 0.01 ）. Conclusions Apollon ASODN may decrease Apollon gene expression, suppress K562 cells proliferation effectively, and induce significant apoptosis of K562 cells. Apollon ASODN is able to reverse the drug resistance via inhibition of Apollon expression and inducement of apoptosis.