反义寡核苷酸阻断雄激素受体表达对前列腺癌细胞生长活性的影响

The growth inhibition effects on prostate cancer cells by blocking expression of androgen receptor with antisense oligonucleotide

目的 探讨反义技术阻断雄激素受体 (AR)基因表达对前列腺癌细胞生长的影响。 方法 设计、合成针对AR基因反义寡核苷酸 (ASODN) ,在脂质体介导下转染前列腺癌细胞。采用RT PCR方法检测AR基因表达 ;MTT比色法检测细胞增殖活性 ;透射电镜、TUNEL检测细胞凋亡 ;流式细胞术分析细胞周期时相。 结果 0 .5～ 2 .0 μmol LASODN作用 12～ 36h ,癌细胞ARmRNA水平降低 10 .45 %～ 82 .10 %(P <0 .0 5 ) ,细胞增殖活性抑制 11.8%～ 5 6 .9%(P <0 .0 5 ) ,细胞周期阻滞于G2 M期 ,部分细胞呈现典型凋亡形态学改变 ,凋亡比率为 34.2 5 %(P <0 .0 1)。 结论 应用ASODN封闭AR基因表达能抑制前列腺癌细胞体外生长活性 ,有望成为前列腺癌基因治疗的有效策略。

ObjectiveTo probe into the growth inhibition effects on prostate cancer cells by antisense oligonucleotide (ASODN) targeting androgen receptor (AR).MethodsThe ASODN for AR was designed and synthesized to transfect prostate cancer cells with liposome.AR expression was assayed by RT-PCR.The cellular proliferation activities were detected by MTT colormetric and flow cytometric assays.The apoptosis was detected by electronic microscopic and TUNEL methods.ResultsWhen transfected with 0.5～2.0 μmol/L ASODN for 12～36 h,the cellular AR mRNA level was decreased by 10.45%～82.10% (P<0.01),with cellular proliferation activities being inhibited by 11.8%～56.9% (P<0.05).Cell cycles were arrested at G 2/M phase with partial cancer cells presenting characteristic changes of apoptosis.The apoptosis rate was 34.25% (P<0.01).ConclusionsThe in vitro growth activities of prostate cancer cells could be inhibited by blocking expression of AR with ASODN,which might be an potential gene therapy strategy of prostate cancer.