PSMAe/p驱动shRNA靶向干扰核干因子治疗前列腺癌的研究

Study of shRNA targeting nucleostemin driven by PSMA enhancer and promoter in prostate cancer therapy

目的探讨前列腺特异性膜抗原增强子、启动子(PSMAe/p)驱动shRNA靶向干扰核干因子(NS)治疗前列腺癌的有效性和应用前景。方法免疫组织化学染色观察NS基因在前列腺癌LNCaP细胞和PC-3细胞中的表达;利用构建的以poly(A)为终止信号的NS特异性shRNA表达载体pPSMAe/p-shNS-poly(A)转染LNCaP细胞和PC-3细胞;观察RNA干扰后细胞体外增殖的变化,检测NS基因水平、细胞周期、细胞凋亡的变化。结果 NS在LNCaP和PC-3细胞中高表达;pPSMA-shNS-poly(A)在转染高表达PSMA的LNCaP细胞后NS表达受到抑制,细胞周期受到阻滞,并抑制了细胞的增殖,同时导致细胞发生凋亡;而在不表达PSMA的PC-3细胞中NS表达无明显抑制,细胞周期未受到阻滞,细胞凋亡无明显变化。结论 PSMAe/p驱动shRNA靶向干扰NS基因具有细胞特异性;NS基因在LNCaP细胞恶性增殖中发挥重要作用,NS可能是前列腺癌基因治疗的理想靶基因之一。

【Objective】 To explore the efficacy and application of shRNA targeting nucleostemin driven by PSMA enhancer and promoter in prostate cancer therapy.【Methods】 Immunohistochemistry was used to detect the expression of NS in human prostate cancer cell lines LNCaP and PC-3.Recombinant plasmid pPSMAe/ p-shNS-poly（A） was transfected into human prostate cell lines LNCaP and PC-3.The changes of cell proliferation ability,NS gene and protein level,cell cycle and apoptosis were also studied after down-regulating the NS gene level.【Results】 Both LNCaP cells and PC-3 cells express NS protein.The expression level of NS gene in LNCaP cells was downregulated,cell cycle was blocked,cell proliferation ability was discounted,cell apoptosis increased;The downregulation of NS gene expression level wasn＇t conspicuous in PC-3 cells,cell cycle and cell proliferation ability didn＇t change obviously.【Conclusion】 The shRNA transcription targeting NS gene driven by PSMAe/p has cellular specificity.NS may act as an important regulator to regulate the malignant proliferation of LNCaP cells.NS gene may serve as an ideal therapeutic target for prostate cancer.