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目的:初步研究前列腺雄激素调节基因(PAR)与雄激素—雄激素受体信号转导通路的关系及其在前列腺癌细胞恶性转化过程中的作用,探讨通过抑制 PAR 基因治疗雄激素非依赖性前列腺癌的可能性。方法:用 RT-PCR 检测 LNCaP、PC3细胞中 PAR 基因 mRNA 表达水平的差异。分别用 RT-PCR 检测双氢睾酮对LNCaP、PC3及稳定转染了 pcDNA3-AR 的 PC3细胞株 PC3-AR 的 PAR 基因 mRNA 表达的调节作用,并观察这一调节作用是否可被雄激素受体拮抗剂氟他胺阻断。进一步用 RNA 干扰技术下调 PC3细胞 PAR 的表达,用细胞计数、软琼脂克隆形成实验、流式细胞术研究 PAR 基囚表达下调对 PC3细胞生长的抑制作用。结果:PC3细胞 PAR 基因 mRNA 的表达是 LNCaP 细胞的3倍;双氢睾酮可调节 LNCaP 和 PC3-AR 细胞株PAR 基因 mRNA 表达水平,此种对 PAR 表达的调节作用可被氟他胺阻断:双氢睾酮对 PC3细胞 PAR 基因mRNA 表达无明显影响。RNA 干扰可抑制 PC3细胞 PAR 基因表达,使细胞增殖受抑制,细胞周期阻滞于G_2-M 期,凋亡增加。结论:PAR 可能是雄激素—雄激素受体信号转导通路下游的与雄激素非依赖性前列腺癌恶性表型密切相关的癌基因,有望成为其基冈和药物治疗的靶点。
OBJECTIVE: To study the relationship between the androgen-androgen receptor (PAR) and androgen-androgen receptor signal transduction pathway and its role in the malignant transformation of prostate cancer cells, and to explore the role of PAR gene in the treatment of androgen-independent Possibility of prostate cancer. Methods: The mRNA expression of PAR gene in LNCaP and PC3 cells was detected by RT-PCR. RT-PCR was used to detect the regulatory effect of dihydrotestosterone on the mRNA expression of PAR gene in PC3-AR cells transfected with pcDNA3-AR, LNCaP, PC3 and pcDNA3-AR, and to observe whether this regulatory effect can be regulated by androgen receptor Flutamide antagonist block. The expression of PAR in PC3 cells was further down-regulated by RNA interference technique. The cytotoxicity against PC3 cells was determined by flow cytometry. Results: The expression of PAR gene mRNA in PC3 cells was 3 times higher than that in LNCaP cells. Dihydrotestosterone could regulate the mRNA expression of PAR gene in LNCaP and PC3-AR cell lines. The modulation of PAR expression could be blocked by flutamide: Dihydrotestosterone had no significant effect on PAR gene mRNA expression in PC3 cells. RNA interference can inhibit the expression of PAR gene in PC3 cells, inhibit cell proliferation, arrest the cell cycle in G2-M phase and increase apoptosis. CONCLUSIONS: PAR may be the oncogene closely related to the androgen-independent prostate cancer malignant phenotype downstream of the androgen-androgen receptor signaling pathway and may be the target of its Kegonia and drug therapy.