雄激素抑制蛋白激酶D1表达促进前列腺癌细胞的侵袭
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摘要
目的研究蛋白激酶D1(PKD1)在雄激素促前列腺癌PC-3细胞侵袭中的作用。方法培养前列腺癌PC-3细胞,选择生长良好的第3～6代细胞,分别以100 nm/L,50 nm/L,10 nm/L睾酮处理后,分为100 n M组、50 n M组和10 n M组,并以包裹空载质粒或PKD1质粒为对照组,Western blot法检测雄激素对PKD1蛋白表达的影响,侵袭小室法检测PKD1在雄激素促PC-3细胞侵袭中的作用。结果与对照组比较,不同浓度的睾酮(10,50,100 nm/L)处理PC-3细胞48 h,均能明显降低PKD1蛋白表达(P<0.05);睾酮(100 nmol/L)能显著促进PC-3细胞的侵袭数目(P<0.001);以PKD1质粒转染PC-3细胞,过表达PKD1后,睾酮促PC-3细胞的侵袭数目明显降低(P<0.001)。结论不同剂量的睾酮可浓度依赖性降低PKD1的蛋白表达,促进前列腺癌细胞侵袭能力。
Objective To investigate the role of Protein Kinase D1( PKD1) in androgen-stimulated prostate cancer cell invasion.Methods The prostate cancer PC3 cells were cultured and the 3 rd to 6 th generation cells with good growth were selected and divided into 100 nM group,500 n M group and 10 nM group with 100 nm/L,50 nm/L and 10 nm/L testosterone respectively.The empty plasmid or PKD1 plasmid was packaged as a control group.Western blot was used to detect the effect of androgen on the expression of PKD1 protein.The invasive compartment assay was used to detect the role of PKD1 in androgen-induced PC-3 cell invasion.Results Compared with the control group,different concentrations of testosterone( 10,50,100 nm/L) for PC-3 cells for 48 hours can significantly reduce the expression of PKD1 protein( P<0.05).The testosterone( 100 nmol/L) significantly promoted the number of invasion of PC-3 cells( P<0. 001).When the PC-3 cells were transfected with PKD1 plasmid and overexpressed PKD1,the number of invasion of PC-3 cells promoted by Testosterone significantly decreased( P<0.001).Conclusion Different doses of testosterone can reduce the protein expression of PKD1 in a concentration-dependent manner and promote the invasion ability of prostate cancer cells.
Objective To investigate the role of Protein Kinase D1( PKD1) in androgen-stimulated prostate cancer cell invasion.Methods The prostate cancer PC3 cells were cultured and the 3 rd to 6 th generation cells with good growth were selected and divided into 100 nM group,500 n M group and 10 nM group with 100 nm/L,50 nm/L and 10 nm/L testosterone respectively.The empty plasmid or PKD1 plasmid was packaged as a control group.Western blot was used to detect the effect of androgen on the expression of PKD1 protein.The invasive compartment assay was used to detect the role of PKD1 in androgen-induced PC-3 cell invasion.Results Compared with the control group,different concentrations of testosterone( 10,50,100 nm/L) for PC-3 cells for 48 hours can significantly reduce the expression of PKD1 protein( P<0.05).The testosterone( 100 nmol/L) significantly promoted the number of invasion of PC-3 cells( P<0. 001).When the PC-3 cells were transfected with PKD1 plasmid and overexpressed PKD1,the number of invasion of PC-3 cells promoted by Testosterone significantly decreased( P<0.001).Conclusion Different doses of testosterone can reduce the protein expression of PKD1 in a concentration-dependent manner and promote the invasion ability of prostate cancer cells.
引文
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[2] Niu Y,Guo C,Wen S,et al.ADT with antiandrogens in prostate cancer induces adverse effect of increasing resistance,neuroendocrine differentiation and tumor metastasis[J].Cancer Lett,2018(439):47-55.
[3] Cai Z,Chen W,Zhang J,et al.Androgen receptor:what we know and what we expect in castration-resistant prostate cancer[J].Int Urol Nephrol,2018,50(10):1 753-1 764.
[4] Steinberg SF.Regulation of protein kinase D1 activity[J].Mol Pharmacol,2012,81(3):284-291.
[5] Ganju A,Chauhan SC,Hafeez BB,et al.Protein kinase D1regulates subcellular localisation and metastatic function of metastasis-associated protein 1[J].Br J Cancer,2018,118(4):587-599.
[6] Zhang D,Zhao S,Li X,et al.Prostate Luminal Progenitor Cells in Development and Cancer[J]. Trends Cancer,2018,4(11):769-783.
[7] Banerjee PP,Banerjee S,Brown TR,et al.Androgen action in prostate function and disease[J].Am J Clin Exp Urol,2018,6(2):62-77.
[8] Bell MA,Campbell JD,Joice G,et al. Shifting the Paradigm of Testosterone Replacement Therapy in Prostate Cancer[J].World J Mens Health,2018,36(2):103-109.
[9] Herati AS,Kohn TP,Butler PR,et al.Effects of Testosterone on Benign and Malignant Conditions of the Prostate[J].Curr Sex Health Rep,2017,9(2):65-73.
[10] Jansen S,Gosens R,Wieland T,et al.Paving the Rho in cancer metastasis:Rho GTPases and beyond[J]. Pharmacol Ther,2018(183):1-21.
[11] Guha S,Tanasanvimon S,Sinnett-Smith J,et al.Role of protein kinase D signaling in pancreatic cancer[J].Biochem Pharmacol,2010,80(12):1 946-1 954.
[12] Sundram V,Chauhan SC,Jaggi M.Emerging roles of protein kinase D1 in cancer[J]. Mol Cancer Res,2011,9(8):985-996.