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目的:利用外科手术去势模拟内分泌治疗,研究Pim-1基因在LNCa P原位前列腺肿瘤裸鼠模型上的表达。方法:利用LNCa P细胞在BALBc裸鼠分别建立雄激素依赖、内分泌治疗和去势抵抗性前列腺肿瘤原位动物模型,采用RT-PCR、qRT-PCR、ELISA和免疫组化等技术检测Pim-1、前列腺特异性抗原(PSA)和雄激素受体(AR)在3组肿瘤组织中的表达差异。结果:RT-PCR结果:电泳结果显示雄激素依赖组与去势抵抗组的相对灰度比值分别为0.59±0.01和1.14±0.02,与内分泌治疗组0.62±0.03比较差异均具有统计学意义(P<0.05);qRT-PCR结果:雄激素依赖组与去势抵抗组的Pim-1△Ct值均分别为6.15±0.34和4.56±0.23,与内分泌治疗组5.11±0.21比较差异均具有统计学意义(P<0.05);应用2-△△Ct相对定量方法分析数据,内分泌治疗组和去势抵抗组Pim-1的扩增产物均较雄激素依赖组分别上调2.05倍和3.01倍;ELSIA结果:空白对照组裸鼠PSA浓度为0μg/L,雄激素依赖组和去势抵抗组均分别为(480±25)、(870±23)pg/ml,与内分泌治疗组(170±32)pg/ml比较差异有统计学意义(P<0.01);免疫组化结果:雄激素依赖组Pim-1和AR的平均光密度比值分别为0.017±0.002和0.032±0.009,去势抵抗组分别为0.024±0.002和0.040±0.011,内分泌治疗组均分别为0.018±0.001和0.019±0.006;雄激素依赖组和去势抵抗组的Pim-1及AR的平均光密度比值均与内分泌治疗组差异有统计学意义(P<0.01)。结论:Pim-1在祼鼠前列腺肿瘤内分泌治疗期呈现出高表达状态,对前列腺肿瘤的发展和转移起很大作用。
OBJECTIVE: To study the expression of Pim-1 gene in nude mouse models of LNCa P in situ prostate cancer by surgical castration simulated endocrine therapy. Methods: The models of androgen-dependent, endocrine and ovariectomized prostate tumor in situ were established by LNCa P cells in BALBc nude mice. RT-PCR, qRT-PCR, ELISA and immunohistochemistry were used to detect the expression of Pim-1 , Prostate-specific antigen (PSA) and androgen receptor (AR) in three groups of tumor tissues. Results: RT-PCR results: The results of electrophoresis showed that the relative gray values of the androgen-dependent group and the castrate-resistant group were 0.59 ± 0.01 and 1.14 ± 0.02, respectively, which were significantly different from those of the endocrine-treated group (P <0.62 ± 0.03) <0.05). The results of qRT-PCR showed that the Pim-1ΔCt values in androgen-dependent group and the castration resistance group were 6.15 ± 0.34 and 4.56 ± 0.23, respectively, which were significantly different from those in the endocrine-treated group (5.11 ± 0.21) (P <0.05). According to the data of 2- △△ Ct relative quantification, the amplification products of Pim-1 in endocrine therapy group and Ovariectomized group were 2.05 times and 3.01 times higher than those in the androgen-dependent group respectively. ELSIA results: The concentration of PSA in blank control group was 0μg / L, and the level of PSA in test group was (480 ± 25) and (870 ± 23) pg / ml in androgen-dependent group and that in endocrine therapy group was 170 ± 32 pg / (P <0.01). The results of immunohistochemistry showed that the average optical density ratios of Pim-1 and AR in androgen-dependent group were 0.017 ± 0.002 and 0.032 ± 0.009, respectively, and those in castration resistance group were 0.024 ± 0.002 and 0.040 ± 0.011, respectively, and the endocrine-treated groups were 0.018 ± 0.001 and 0.019 ± 0.006, respectively. In the androgen-dependent group and the castrate-resistant group, Pim The mean optical density ratio of -1 and AR were significantly different from that of endocrine group (P <0.01). CONCLUSION: Pim-1 is highly expressed in the endocrine stage of zoonotic prostate tumors and plays a significant role in the development and metastasis of prostate tumors.