乳胞素诱导前列腺癌细胞凋亡机制的初步研究  

Investigation of the apoptosis mechanism induced by lactacystin in prostate cancer cell

作　　者：高海峰[1] 王彦[2] 李天明[1] 李英华[3] 于广海[1] 

机构地区：[1]大连市中心医院泌尿外科,116033 [2]大连医科大学附属第二医院实验中心,116033 [3]大连医科大学附属第二医院肿瘤内科,116033

出　　处：《中国医师进修杂志》2012年第32期1-4,共4页Chinese Journal of Postgraduates of Medicine

摘　　要：目的探讨乳胞素在诱导前列腺癌细胞凋亡作用中与核转录因子（NF）-κB活性的关系。方法利用乳胞素作用两种前列腺癌细胞，设立培养液处理的空白对照组和乳胞素处理组，乳胞素处理组分别向两种细胞中加入浓度递增的乳胞素（0．5、1．0、2．0、4．0μmol/L），作用时间分别为8、16和24h。MTT法检测细胞生存率，利用酶联免疫吸附试验定量分析NF-κB DNA结合活性，Western blot法检测NF-κB p65核蛋白表达，酶分析法测定caspase-3活性。结果DU145细胞基础状态下NF-κB DNA结合活性强于LNCaP细胞（t=4．728，P=0．001），应用乳胞素作用24h后，与空白对照组比较，不同浓度乳胞素处理组均观察到NF—κB DNA结合活性减少。随着乳胞素浓度增加，LNCaP细胞NF-κB p65核蛋白表达水平下调，而DU145细胞NF-κB p65核蛋白表达水平没有变化。DU145细胞基础状态下caspase-3活性强于LNCaP细胞（t=4．519，P=0．001），乳胞素作用24h后，DU145和LNCaP细胞caspase-3活性均随乳胞素浓度增加升高（2．0μmol／L乳胞素处理组与1．0μmol／L乳胞素处理组比较，DU145细胞P=0．000，LNCaP细胞P=0．000）。结论乳胞素对不同前列腺癌细胞有不同杀伤作用，并与抑制NF-κB活性促进前列腺癌细胞凋亡相关，对激素非依赖性前列腺癌细胞还可能存在其他抗肿瘤细胞生存途径。Objective To investigate the relationship between nuclear factor （NF）-κB activity and lactacystin induced prostate cancer cell apoptosis. Methods Two prostate cancer cell were divided into two groups： blank control group treated with culture solution, lactacystin group treated with different concentration of lactacystin （0.5,1.0,2.0,4.0 μmol/L） ,the action time were 8,16 and 24 hours. The cell survival rate was measured by MTT assay. NF- κB DNA binding activity was measured by enzyme-linked immunosorhent assay, the expression of NF-κB p65 nuclear protein was detected by Western blot assay, and easpase-3 activity was analyzed by enzyme analysis assay. Results On basal condition,the NF-κB DNA binding activity was much higher in DU145 cell than that in LNCaP cell （t = 4.728 ,P = 0.001 ）. Compared with blank control group,different concentration of lactacystin groups＇ NF-K B DNA binding activity in both the LNCaP and DU145 cell were reduced. The expression of NF-κB p65 nuclear protein decreased along with raising of lactaeystin concentration in LNCaP cell, but it did not change in DU 145 cell. On basal condition, caspase-3 activity in DU145 cell was higher than that in LNCaP cell （t = 4.519 ,P = 0.001 ）. After lactaeystin acting of 24 hours, caspase-3 activity increased along with raising of lactacystin concentration in both the LNCaP and DU145 cell （2.0 ~ mol/L lactacystin group compared with 1.0 μmol/L lactacystin group, DU145 cell P = 0.000, LNCaP cell P = 0.000）. Conclusions Lactacystin has different killing effects on prostate cancer cell. The mechanism may be related to inducing the apoptosis by down-regulation of NF-κB activity. There may be additional cell survival/death pathway in androgen-independent prostate cancer cell.

关 键 词：前列腺肿瘤 NF-ΚB 乳胞素 

分 类 号：R737.25[医药卫生—肿瘤]