多西紫杉醇诱导前列腺癌PC-3细胞自噬的研究

发布时间：2018-10-19 07:14

【摘要】：目的：前列腺癌（prostate cancer，PC）是欧美等发达国家和地区男性中最常见的泌尿系统恶性肿瘤。近年来在我国其发病率也呈增长趋势。前列腺癌的治疗包括根治性前列腺切除术、内分泌治疗、化疗等综合治疗方案。由于前列腺癌是一种激素依赖性肿瘤，内分泌治疗是晚期前列腺癌的首选治疗方案。但是绝大多数前列腺癌在1-2年的治疗中位期后会逐渐转变为激素非依赖性，而对激素非依赖性前列腺癌目前尚无更多有效治疗方案，而化疗则是可以选择的方法之一。多西紫杉醇是目前临床上使用的一线化疗药物，具有广泛的抗肿瘤活性，，但后期会逐渐出现化疗抵抗，使疗效变差。化疗抵抗是一个多因素参与的过程，而自噬是新近发现的化疗抵抗的重要机制之一。自噬也被称为Ⅱ型细胞程序性死亡，广泛存在于真核细胞中，参与细胞生长调控，维持细胞内环境稳定。有研究表明，自噬与肿瘤的发生发展有一定的关系。多西紫杉醇作为紫杉类药物，在胃癌，宫颈癌等实体肿瘤中有着广泛应用，也是前列腺癌的主要化疗药物，但关于多西紫杉醇是否可以诱导前列腺癌细胞发生自噬，目前尚未见文献报道。本实验拟通过体外细胞实验，观察多西紫杉醇是否诱导前列腺癌PC-3细胞产生自噬，并初步探讨其可能机制，为临床应用提供理论和实验依据。 方法：不同浓度多西紫杉醇作用于前列腺癌PC-3细胞24h后，应用MTT法测定各组细胞的增殖率；透射电镜观察是否有自噬体的形成；免疫荧光和Western blot分别定性和定量检测自噬相关蛋白Ⅱ型微管相关蛋白轻链3蛋白(microtubule-associated protein1light chain3typeⅡ，LC3-Ⅱ)，底物蛋白(sequestosome1/SQSTM1，P62)表达水平，RT-PCR检测自噬相关基因Beclin-1mRNA。 结果：MTT结果显示多西紫杉醇能有效抑制PC-3细胞增殖(F=58.684,P=0.000)；10-6mol/L多西紫杉醇处理PC-3细胞24h可诱导其发生明显的自噬，透射电镜观察到自噬体双层膜结构的形成。免疫荧光和Western blot结果显示多西紫杉醇处理PC-3细胞后P62表达降低(P=0.003、P=0.000、P=0.000)，LC3-Ⅱ表达增强(P=0.002、P=0.000、P=0.000)，RT-PCR结果显示多西紫杉醇处理PC-3细胞后可引起细胞Beclin-1mRNA水平的上调（P=0.000、P=0.000、P=0.000）。 结论：本实验证实多西紫杉醇能有效抑制前列腺癌PC-3细胞的增殖并诱导其发生自噬，其作用机制可能与上调Beclin-1的mRNA水平有关。
[Abstract]:Objective: prostate cancer (prostate cancer,PC) is the most common malignant tumor of urinary system in men in developed countries and regions. In recent years, the incidence of disease is also increasing in China. Prostate cancer treatment includes radical prostatectomy, endocrine therapy, chemotherapy and other comprehensive treatment. As prostate cancer is a hormone-dependent tumor, endocrine therapy is the preferred treatment for advanced prostate cancer. But the majority of prostate cancer will gradually change to hormone independent after 1-2 years of median treatment, and there is no more effective treatment for hormone independent prostate cancer, and chemotherapy is one of the options. Docetaxel is a first-line chemotherapeutic drug currently used in clinic. It has a wide range of antitumor activities, but chemotherapeutic resistance will gradually appear in the later stage, which makes the curative effect worse. Chemotherapeutic resistance is a multifactorial process, and autophagy is one of the important mechanisms of chemotherapeutic resistance. Autophagy, also known as type II cell programmed death, is widely present in eukaryotic cells, participates in the regulation of cell growth and maintains the stability of the intracellular environment. Studies have shown that autophagy has a certain relationship with the occurrence and development of tumor. Docetaxel, as a taxol, is widely used in solid tumors such as gastric cancer, cervical cancer and other solid tumors. It is also the main chemotherapeutic drug for prostate cancer, but whether docetaxel can induce autophagy in prostate cancer cells. At present, there is no literature report. The aim of this study was to investigate whether docetaxel induces autophagy in prostate cancer PC-3 cells in vitro and to explore its possible mechanism, which provides theoretical and experimental basis for clinical application. Methods: after treated with different concentrations of docetaxel for 24 hours, the proliferation rate of prostate cancer PC-3 cells was measured by MTT assay, and the formation of autophagy was observed by transmission electron microscope. Microtubule-associated protein1light chain3type 鈪

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