Wang Jiang, Li Gang, Li Bo, Song Hualin, Shang Zhiqun, Jiang Ning, Niu Yuanjie
Department of Urology, Tianjin Institute of Urology, The Second Hospital of Tianjin Medical University, Tianjin 300211, P.R. China.
Tianjin Municipal Research Institute for Family Planning, Tianjin 300131, P.R. China.
Oncol Lett. 2017 Jun;13(6):4364-4370. doi: 10.3892/ol.2017.6010. Epub 2017 Apr 7.
The aim of the present study was to observe the dynamic changes of proto-oncogene, serine/threonine kinase, Pim-1 at the gene and protein level in a mouse model of prostate cancer following surgical castration. Using LNCaP cells to establish a subcutaneous xenograft model and orthotopic prostate cancer BALB/c nude mouse models, the xenograft models were divided into an androgen-dependent prostate cancer group (ADPC), an androgen deprivation therapy (ADT) group and an androgen independent prostate cancer (AIPC) group. Reverse transcription-polymerase chain reaction (RT-PCR), RT-quantitative PCR, ELISA and immunohistochemistry analyses were performed to compare the expression levels of Pim-1, prostate-specific antigen (PSA) and androgen receptor (AR) in tumor tissue of three subgroups. Agarose gel electrophoresis revealed that the RT-PCR results of the ADPC (0.59±0.01) and AIPC groups (1.14±0.015) were significantly different when compared with the ADT group (0.62±0.026; P<0.05). As for RT-qPCR, the ΔCq of Pim-1 in the ADPC (6.15±0.34) and AIPC (4.56±0.23) groups were significantly different compared with the ADT group (5.11±0.21; P<0.05). Using 2 as a relative quantification method to analyze the data, the amplification products of Pim-1 increased by 2.05 and 3.01 times in the ADT and AIPC groups, respectively. ELISA demonstrated the following: The serum concentration of PSA was 0 ng/ml in the control group, 0.48±0.025 ng/ml in the ADPC group and 0.87±0.023 ng/ml in the AIPC group, which were significantly different compared with the ADT group (0.17±0.032 ng/ml; P<0.01). Upon immunohistochemical staining, the protein expression levels of Pim-1 and AR, respectively, were 0.017±0.0021 and 0.032±0.009 in the ADPC group, 0.024±0.0019 and 0.040±0.011 in the AIPC group, and 0.018±0.0013 and 0.019±0.006 in the ADT group. The protein levels of Pim-1 and AR in the ADPC and AIPC groups were significantly different compared with the ADT group (P<0.01). In addition, an orthotopic prostate cancer animal model of ADT was successfully established in the current study, and further investigation revealed that ADT did not affect the expression of Pim-1 at the gene or protein levels; thus, it is hypothesized that Pim-1 may be important in the proliferation and differentiation of prostate cancer during ADT.
本研究的目的是观察去势手术前列腺癌小鼠模型中原癌基因丝氨酸/苏氨酸激酶Pim-1在基因和蛋白质水平的动态变化。利用LNCaP细胞建立皮下异种移植模型和原位前列腺癌BALB/c裸鼠模型，将异种移植模型分为雄激素依赖性前列腺癌组（ADPC）、雄激素剥夺治疗（ADT）组和雄激素非依赖性前列腺癌（AIPC）组。采用逆转录-聚合酶链反应（RT-PCR）、RT-定量PCR、酶联免疫吸附测定（ELISA）和免疫组织化学分析比较三个亚组肿瘤组织中Pim-1、前列腺特异性抗原（PSA）和雄激素受体（AR）的表达水平。琼脂糖凝胶电泳显示，与ADT组（0.62±0.026）相比，ADPC组（0.59±0.01）和AIPC组（1.14±0.015）的RT-PCR结果有显著差异（P<0.05）。至于RT-qPCR，ADPC组（6.15±0.34）和AIPC组（4.56±0.23）中Pim-1的ΔCq与ADT组（5.11±0.21）相比有显著差异（P<0.05）。以2为相对定量方法分析数据，ADT组和AIPC组中Pim-1的扩增产物分别增加了2.05倍和3.01倍。ELISA结果如下：对照组血清PSA浓度为0 ng/ml，ADPC组为0.48±0.025 ng/ml，AIPC组为0.87±0.023 ng/ml，与ADT组（0.17±0.032 ng/ml）相比有显著差异（P<0.01）。免疫组织化学染色显示，ADPC组中Pim-1和AR的蛋白质表达水平分别为0.017±0.0021和0.032±0.009，AIPC组为0.024±0.0019和0.040±0.011，ADT组为0.018±0.0013和0.019±0.006。ADPC组和AIPC组中Pim-1和AR的蛋白质水平与ADT组相比有显著差异（P<0.01）。此外，本研究成功建立了ADT原位前列腺癌动物模型，进一步研究发现ADT不影响Pim-1在基因或蛋白质水平的表达；因此，推测Pim-1在ADT期间前列腺癌的增殖和分化中可能起重要作用。
