比卡鲁胺抑制CWR22Rv1细胞移植裸鼠前列腺癌的机制研究

Inhibitory Effects of Bicalutamide on Growth of CWR22Rv1(22Rv1) Cell Transplanted Prostate Cancer in Nude Mice

目的:探讨比卡鲁胺抗CWR22Rv1细胞移植裸鼠前列腺癌的作用机制。方法:采用瘤块接种的方法制作裸小鼠前列腺癌模型,观察比卡鲁胺治疗后的移植瘤重量和体积,并计算抑瘤率,收集移植瘤标本,使用光镜观察法、免疫组化法、基因芯片法和实时定量RT-PCR法比较比卡鲁胺组和阴性对照组肿瘤相关指标的差异。结果:①比卡鲁胺低、中、高剂量组抑瘤率分别为10.77%、37.32%和63.28%(P<0.01)。②光镜观察:对照组肿瘤内癌细胞生长旺盛,移植癌细胞保持原有异型性。而比卡鲁胺低、中和高剂量组呈腺癌典型表现,癌细胞出现不同程度的退变。③免疫组化:低、中、高剂量组增殖细胞核抗原增殖指数同阴性对照组相比明显减小(P<0.01)。对照组每200个细胞中平均有180±8个细胞雄激素受体(androgen receptor,AR)呈阳性表达,而高剂量组中AR阳性细胞数较对照组明显减少(P<0.01)。④基因芯片:高剂量组在前列腺癌相关基因表达丰度上明显低于对照组,主要为AR和关联蛋白、细胞黏附分子和细胞周期相关基因,并且实时定量RT-PCR结果与基因芯片结果基本一致。结论:比卡鲁胺抗22Rv1细胞移植裸鼠前列腺癌效果显著,主要通过降低细胞核AR表达、减少细胞黏附、调控细胞周期和抑制细胞增殖等发挥作用。

Objective： To study the effect ofbicalutamide on growth of transplanted prostate cancer using CWR22Rv1 cell in nude mice and corresponding mechanism. Methods： Prostate cancer was made by having transplanted tumor tissue into the nude mice. The volume, weight and the inhibition rate of the tumor in nude mice were calculated after treating by bicalutamidc for 30 d, subcutaneous tumors were subjected to histological examination, immunohistochemistry, cDNA microarray technique and Real-time PCR. Results： 1） In every treatment group tumor growth was suppressed significantly （P〈0.01）. The inhibition rate of the low, middle, high dose of bicalutamide was 10.77 %, 37.32 %, and 63.28 %, respectively. 2） Light microscopy： Cancer cells arc cugonic in control group, and there were different degrees of hetcromophism in the transplanted cancer cells. However, the cancer cells of low, middle, high dose of bicalutamide groups showed a typical adcnocarcinoma with different degrees of degeneration. 3） Immunohistochemistry： PCNA of the low, middle, high dose of bicalutamide was markedly lower than that of the control （P〈0.01）. 4） cDNA microarray： Prostate cancer-related genes were down-regulated in the high dose bicalutamide group and could be categorized： androgen receptor and associated proteins, cell adhesion and cell cycle. The result of Real-time PCR was generally consistent with the cDNA microarray result. Conclusion： Bicalutamide can significantly inhibit the growth of transplanted prostate cancer using 22Rv1 cell in nude mice in vivo by inhibiting cell proliferation, reducing the AR expression, blocking cell adhesion and regulating cell cycle.