双氢睾酮和氟他胺对前列腺癌LNCaP细胞内钙离子的影响
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摘要
目的:观察双氢睾酮(DHT)和氟他胺(FLU)作用于雄激素依赖性前列腺癌细胞LNCa P内Ca~(2+)的变化,探讨DHT和FLU引起LNCa P细胞内Ca~(2+)变化的作用机制。方法:获得雄激素依赖性(LNCa P)细胞株,进行细胞培养,获得稳定生长及传代的细胞株。不同浓度的雄激素受体激动剂DHT及雄激素受体拮抗剂FLU作用于培养的LNCa P细胞株,不同时间点取干预措施下的LNCa P细胞,通过荧光染料负载,经流式细胞术检测并比较不同时间点、不同浓度药物作用下LNCa P细胞Ca~(2+)的变化和差异。结果:DHT作用于LNCa P细胞后,在每个剂量组中Ca~(2+)荧光强度差异有统计学意义(P<0.05),不同时间点变化差异有统计学意义(P<0.05)。FLU干预LNCa P后,各个剂量组的Ca~(2+)荧光强度随着时间的变化差异有统计学意义(P<0.05)。结论:DHT降低LNCa P细胞内Ca~(2+)含量不明显,说明对LNCa P细胞内的信号转导及凋亡影响不大;而FLU可明显提高LNCa P细胞内Ca~(2+)的浓度,导致钙超载,促进细胞的凋亡。
Objective: To investigate the change of Ca~(2 +) and discuss the mechanism of Ca~(2 +) changes in LNCa P cells by DHT and FLU. Methods: To abtain drogen- dependent( LNCa P) prostate cancer cell lines,by cell culture,to abtain stable cell strains. The androgen receptor agonist DHT and the androgen receptor antagonist FLU acted on the culture prostate cancer cell line of androgen at different concentrations drugs. We extracted cells at different time points of LNCa P under the interventions. By fluorescent dye load measured,to compare the Ca~(2 +) differences at different time points and under the action of different drug concentrations in LNCa P by flow cytometry. Results: After DHT acts on LNCa P cells,the Ca~(2 +) of LNCa P increased significantly( P < 0. 05),but the difference of the increasing degree of each index in different time points. After FLU acts on LNCa P cells,the content of Ca~(2 +) increased significantly( P <0. 05). Conclusion: The content of Ca~(2 +) is not obvious decreased in LNCa P cell by DHT,showed the low effect on signal transduction and the apoptosis. However,the concentration of Ca~(2 +) in LNCa P cells could be increased obviously by FLU,which leads to calcium overload and promotes cell apoptosis.
Objective: To investigate the change of Ca~(2 +) and discuss the mechanism of Ca~(2 +) changes in LNCa P cells by DHT and FLU. Methods: To abtain drogen- dependent( LNCa P) prostate cancer cell lines,by cell culture,to abtain stable cell strains. The androgen receptor agonist DHT and the androgen receptor antagonist FLU acted on the culture prostate cancer cell line of androgen at different concentrations drugs. We extracted cells at different time points of LNCa P under the interventions. By fluorescent dye load measured,to compare the Ca~(2 +) differences at different time points and under the action of different drug concentrations in LNCa P by flow cytometry. Results: After DHT acts on LNCa P cells,the Ca~(2 +) of LNCa P increased significantly( P < 0. 05),but the difference of the increasing degree of each index in different time points. After FLU acts on LNCa P cells,the content of Ca~(2 +) increased significantly( P <0. 05). Conclusion: The content of Ca~(2 +) is not obvious decreased in LNCa P cell by DHT,showed the low effect on signal transduction and the apoptosis. However,the concentration of Ca~(2 +) in LNCa P cells could be increased obviously by FLU,which leads to calcium overload and promotes cell apoptosis.
引文
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[3]Monge A,Jagla M,Lapouge G,et al.Unfaith flulness and promiscuity of a mutant androgen receptor in a hormone-refractory prostate cancer[J].Cell Mol Life Sci,2006,63(4):487-497.
[4]Wang Y.Change of the cell cycle after flutamide treatment in prostate cancer cells and its molecular mechanism[J].Asian J Androl,2005,7(4):375-380.
[5]Ostadal B.Developmental determinants of cardiac sensitivity to hypoxia[J].Can J Physiol Pharmacol,2014,92(7):566-574.
[6]Le TH,Nguyen NT.A complete mitochondrial genome from echinochasmus japonicus supports the elevation of echinochasminae Odhner,1910 to family rank(Trematoda:Platyhelminthes)[J].Infect Genet Evol,2016,45:369-377.
[7]Hirlee T,Tutaka S,Yuanbin L,et al.There gulation of reactive oxgens peeies production and program medecil dealth[J].Cell Biol,1998,141(6):1423-1432.
[8]Ozaki T.Inhibitory peptide of mitochondrial u-calpain protects against photoreceptor degeneration in rhodopsin transgenic S334ter and P23H rats[J].PLo S One,2013,8(8):1-10.
[9]Xiao T.Activation of an apoptotic signal transduction pathway involved in the upregulation of calpain and apoptosis inducing factor in aldosterone induced primary cultured cardiomyocytes[J].Food Chem Toxicol,2013,53:364-370.
[10]Paredes RM,Etzler JC.Chemical calcium indicators[J].Methods,2008,46(3):143-151.
[11]Shkryl VM,Shirokova N.Transfer and tunneling of Ca2+from sarcoplasmic reticulum to mitochondria in skeletal muscle[J].Biol Chem,2006,281(3):1547-1554.
[12]Tsai MO,Malley B.Molecular mechanisms of action of steroid/thyroid receptor super family members[J].Annu Rev Biochem,1994,63:451-486.
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