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摘要:
目的 探讨多西紫杉醇治疗男性雄激素抵抗前列腺癌(castration-resistant prostate cancer, CRPC)的分子机制。 方法 前列腺癌细胞株LNCaP、PC-3和CW22-rv1体外培养后, 通过蛋白质印迹、细胞转染、荧光素酶分析、细胞存活率分析等试验分析多西紫杉醇(Docetaxel, Doc)处理后细胞株的存活、AR及p-c-jun的表达情况及其与细胞生存的关系, 同时实时定量PCR检测相应mRNA的表达情况。 结果 多西紫杉醇对不同前列腺癌细胞株的敏感性不同, 其中PC-3细胞最敏感, CW22-rv1和LNCaP细胞中度敏感, 其敏感性与p-c-jun表达呈负相关。转染c-jun基因可降低细胞对多烯紫杉醇的敏感性, 而PC-3细胞转染c-jun和AR基因后则可以使细胞恢复到中等程度敏感性, 细胞存活率为30%。长期暴露于Bicalutmide(比卡鲁胺)后的LNCaP细胞经Doc处理后PSA蛋白表达增加, AR蛋白表达水平降低, AR的mRNA却增加。 结论 p-c-jun降低前列腺癌细胞株对多西紫杉醇的敏感性, 而AR可以增加前列腺癌细胞株对多西紫杉醇的反应, AR的上调降低c-jun/p-c-jun的转录活性, 从而增加前列腺癌细胞株对Doc的反应, 可能是Doc治疗前列腺癌的机制之一。 -
关键词:
- 雄激素受体(AR) /
- p-c-jun /
- 多西紫杉醇(Doc) /
- 雄激素抵抗前列腺癌(CRPC) /
- 前列腺癌细胞
Abstract:Objective The present study investigated the biochemical mechanism of docetaxel(Doc) for the treatment of castration-resistant prostate cancer(CRPC). Methods The prostate cancer cell lines PC-3, LNCaP, and CW22-rv1 were grown in vitro.The expression of c-Jun and the androgen receptor(AR), the relationship between their expression levels, and the viability of LNCaP, PC-3, and CW22-rv1 cells after exposure to Doc were studied in vitro by immunoblot, luciferase, and viability assays.Furthermore, the expression of AR mRNA was analyzed by quantitative real-time polymerase chain reaction(RT-PCR). Results Doc therapy elicited different responses in the different cell lines.PC-3 cells were the most sensitive to treatment, whereas a moderate response was observed in CW22-rv1 and LNCaP cells.The sensitivity and response to Doc was negatively correlated with the level of phosphorylated c-Jun(p-c-Jun) expression.Transfection of the c-jun gene into cell lines decreased their response to Doc treatment, whereas cotransfection of the c-jun and AR genes restored the sensitivity of PC-3 to a moderate degree, with 30% viability was 30%.After long-term exposure to bicalutamide(cas), the Doc-treated LNCaP cells demonstrated increased levels of the prostate specific antigen(PSA).Likewise the AR protein levels decreased with increasing AR mRNA expression. Conclusion This study demonstrated that the level of p-c-Jun decreased the response of PC cells to Doc treatment, but the level of AR increases PC cells response to Doc treatment.The increased AR level inhibits c-Jun/p-c-Jun transcriptional, which decreases the protective effects of Doc in PC cells. -
Key words:
- androgen receptor /
- p-c-jun /
- docetaxel /
- castration-resistant prostate cancer /
- prostate cancer cells
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图 1 三种前列腺癌细胞暴露于5 n M Doc培养0~30 d
Figure 1. PC-3, CW22-rv1, and LNCa P cell lines treated with 5 n M Doc for 0 d to 30 d
图 2 三种细胞暴露于5 n M Doc培养24、48 h后存活率
Figure 2. Percentage of viable cells in PC-3, CW22-rv1, and LNCa P af-ter exposure to 5 n M Doc for 24 and 48 h
图 3 PC-3细胞转染c-jun和AR基因后对Doc的敏感性
Figure 3. Sensitivity to Doc in PC-3 cells transfected with c-jun and AR
图 4 LNCa P-bic及LNCa P细胞暴露于多西紫杉醇(Doc, 5 n M), 双氢睾酮(DHT, 10 n M)或比卡鲁胺(cas, 10μM)24 h后PSA、AR表达的变化
Figure 4. Expression of PSA and AR in LNCa P-bic and LNCa P cells af-ter exposure to Doc(5 n M), DHT(10 n M), or cas(10μM)for 24 h, as well as cells without Doc treatment(0 h)
图 5 四种细胞暴露于5 n M Doc培养24、48h后的细胞存活率
Figure 5. Percentage of viable cells in PC-3, CW22-rv1, LNCa P, and LNCa P-bic cells after exposure to 5 n M Doc for 24 and 48 h
图 6 AP-1转染LNCa P和LNCa P-bic细胞后, 不同处理对AP-1的转录活性的影响
Figure 6. Transcriptional activity of AP-1 after transfection with AP-1-luc and treatment with 5 n M Doc, 10 n M DHT, or 10 n M TPA
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[1] Jemal A, Siegel R, Ward E, et al. Thun MJ. Cancer statistics[J]. CACancer J Clin, 2007, 7(57): 43–46. [2] 叶定伟, 李长岭. 前列腺癌发病趋势的回顾和展望[J]. 中国癌症杂志, 2007, 17(3): 177-180. doi: 10.3969/j.issn.1007-3639.2007.03.001 [3] 周智恩, 李汉忠, 严维刚. 雄激素受体蛋白表达及其基因获得/缺失与前列腺癌预后的相关性[J]. 基础医学与临床, 2010, 30(8): 857-861. https://www.cnki.com.cn/Article/CJFDTOTAL-JCYL201008024.htm [4] Lopergolo A, Zaffaroni N. Biomolecular markers of outcome pre diction in prostate cancer[J]. Cancer, 2009, 115(13 Suppl): 3058-3067. [5] 刘征, 杨为民, 叶章群, 等. 紫杉醇联合肿瘤坏死因子相关凋亡诱导配体对前列腺癌细胞的抑制作用[J]. 华中科技大学学报(医学版), 2007, 36(3): 352-354. doi: 10.3870/j.issn.1672-0741.2007.03.020 [6] Jiang J, Huang H. Targeting the Androgen Receptor by Taxol inCastration-Resistant Prostate Cancer[J]. Mol Cell Pharmacol, 2010, 2(1): 1-5. [7] Petrylak DP, Tangen CM, Hussain MH, et al. Docetaxel and estra mustine compared with mitoxantrone and prednisone for advancedrefractory prostate cancer[J]. N Engl J Med, 2004, 351(15): 1513-1520. doi: 10.1056/NEJMoa041318 [8] McKeage K. Docetaxel: a review of its use for the first-line treat ment of advanced castration-resistant prostate cancer[J]. Drugs, 2012, 72(11): 1559-1577. doi: 10.2165/11209660-000000000-00000 [9] Shapiro D, Tareen B. Current and emerging treatments in the man agement of castration-resistant prostate cancer[J]. Curr Drug Tar gets, 2012, 12(7): 951-964. [10] 宋登新, 陈安民, 郭风劲, 等. siRNA抑制人高迁移率族蛋白1表达对前列腺癌PC-3增殖的影响[J]. 中国肿瘤临床, 2008, 35(6): 339-343. [11] 刘荣华, 王世宣, 马湘一, 等. 二肽基肽酶对前列腺癌细胞系1E8侵袭转移的影响[J]. 中国肿瘤临床, 2008, 35(17): 1003-1006. [12] Pfahl M. Nuclear receptor/Ap-1 interaction[J]. Endocr Rev, 1993, 14(5): 651-658. [13] Hemmerlein B, Weseloh RM, Mello de Queiroz F, et al.0verexpres sion of Eagl potassium channels in clinical tumours[J]. Mol Cancer, 2006, 5(1): 41-54. doi: 10.1186/1476-4598-5-41 [14] O'Neill A J, Prencipe M, Dowling C, et al. Characterisation and ma nipulation of docetaxel resistant prostate cancer cell lines[J]. Molecu lar Cancer, 2011, 10: 126. doi: 10.1186/1476-4598-10-126 [15] Lamont KR, Tindall DJ. Androgen regulation of gene expression[J]. Adv Cancer Res, 2010, 107: 137-162. [16] Potapova O, Basu S, Mercola D, et al. Protective role for c-Jun inthe cellular response to DNA damage[J]. J Biol Chem, 2001, 276(30): 28546-28553. doi: 10.1074/jbc.M102075200 -
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