【摘要】：目的：前列腺癌（prostate cancer，PC）是歐美等發(fā)達(dá)國(guó)家和地區(qū)男性中最常見的泌尿系統(tǒng)惡性腫瘤。近年來在我國(guó)其發(fā)病率也呈增長(zhǎng)趨勢(shì)。前列腺癌的治療包括根治性前列腺切除術(shù)、內(nèi)分泌治療、化療等綜合治療方案。由于前列腺癌是一種激素依賴性腫瘤，內(nèi)分泌治療是晚期前列腺癌的首選治療方案。但是絕大多數(shù)前列腺癌在1-2年的治療中位期后會(huì)逐漸轉(zhuǎn)變?yōu)榧に胤且蕾囆�，而�?duì)激素非依賴性前列腺癌目前尚無更多有效治療方案，而化療則是可以選擇的方法之一。多西紫杉醇是目前臨床上使用的一線化療藥物，具有廣泛的抗腫瘤活性，，但后期會(huì)逐漸出現(xiàn)化療抵抗，使療效變差。化療抵抗是一個(gè)多因素參與的過程，而自噬是新近發(fā)現(xiàn)的化療抵抗的重要機(jī)制之一。自噬也被稱為Ⅱ型細(xì)胞程序性死亡，廣泛存在于真核細(xì)胞中，參與細(xì)胞生長(zhǎng)調(diào)控，維持細(xì)胞內(nèi)環(huán)境穩(wěn)定。有研究表明，自噬與腫瘤的發(fā)生發(fā)展有一定的關(guān)系。多西紫杉醇作為紫杉類藥物，在胃癌，宮頸癌等實(shí)體腫瘤中有著廣泛應(yīng)用，也是前列腺癌的主要化療藥物，但關(guān)于多西紫杉醇是否可以誘導(dǎo)前列腺癌細(xì)胞發(fā)生自噬，目前尚未見文獻(xiàn)報(bào)道。本實(shí)驗(yàn)擬通過體外細(xì)胞實(shí)驗(yàn)，觀察多西紫杉醇是否誘導(dǎo)前列腺癌PC-3細(xì)胞產(chǎn)生自噬，并初步探討其可能機(jī)制，為臨床應(yīng)用提供理論和實(shí)驗(yàn)依據(jù)。 方法：不同濃度多西紫杉醇作用于前列腺癌PC-3細(xì)胞24h后，應(yīng)用MTT法測(cè)定各組細(xì)胞的增殖率；透射電鏡觀察是否有自噬體的形成；免疫熒光和Western blot分別定性和定量檢測(cè)自噬相關(guān)蛋白Ⅱ型微管相關(guān)蛋白輕鏈3蛋白(microtubule-associated protein1light chain3typeⅡ，LC3-Ⅱ)，底物蛋白(sequestosome1/SQSTM1，P62)表達(dá)水平，RT-PCR檢測(cè)自噬相關(guān)基因Beclin-1mRNA。 結(jié)果：MTT結(jié)果顯示多西紫杉醇能有效抑制PC-3細(xì)胞增殖(F=58.684,P=0.000)；10-6mol/L多西紫杉醇處理PC-3細(xì)胞24h可誘導(dǎo)其發(fā)生明顯的自噬，透射電鏡觀察到自噬體雙層膜結(jié)構(gòu)的形成。免疫熒光和Western blot結(jié)果顯示多西紫杉醇處理PC-3細(xì)胞后P62表達(dá)降低(P=0.003、P=0.000、P=0.000)，LC3-Ⅱ表達(dá)增強(qiáng)(P=0.002、P=0.000、P=0.000)，RT-PCR結(jié)果顯示多西紫杉醇處理PC-3細(xì)胞后可引起細(xì)胞Beclin-1mRNA水平的上調(diào)（P=0.000、P=0.000、P=0.000）。 結(jié)論：本實(shí)驗(yàn)證實(shí)多西紫杉醇能有效抑制前列腺癌PC-3細(xì)胞的增殖并誘導(dǎo)其發(fā)生自噬，其作用機(jī)制可能與上調(diào)Beclin-1的mRNA水平有關(guān)。
[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

本文編號(hào)：2280500