淫羊藿素对BALB/c-nu裸鼠前列腺癌组织磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)信号通路及E-钙黏蛋白的影响

摘要

目的 观察淫羊藿素对雄激素依赖型前列腺癌模型小鼠前列腺癌组织磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)信号通路及E-钙黏蛋白的影响并探讨其机制.方法 雄性BALB/c-nu裸鼠40只,随机分为空白对照组、荷瘤组、LY294002组和淫羊藿素组,每组10只,除空白对照组外均采用含有2×107个/mL的人LNCaP前列腺癌细胞悬液前列腺内注射以建立前列腺癌LNCaP荷瘤鼠模型.2周后淫羊藿素组按80 mg/kg剂量尾静脉注射淫羊藿素治疗4周,PI3K/Akt抑制剂组尾静脉注射PI3K/Akt抑制剂LY294002,空白对照组、荷瘤组尾静脉注射等体积生理盐水.治疗结束后取前列腺组织,采用Western blotting检测磷酸化雄激素受体AR(p-AR)、磷酸化蛋白激酶(p-Akt)、雄激素受体剪接变异体7(AR-V7);免疫荧光双重染色(FITC-CY3)法检测降钙素(Calcitonin)和钙黏蛋白E(E-cadherin)表达.结果 荷瘤组前列腺组织p-AR和p-Akt均高表达,前列腺瘤组织芯片标本中AR-V7和Calctionin阳性率分别为(32.45±2.15)％和(46.31±5.01)％,E-cadherin阳性率(22.93±1.96)％,与空白对照组比较差异有统计学意义(P＜0.05).淫羊藿素组前列腺组织p-AR和p-Akt低表达,AR-V7和Calctionin阳性率分别为(17.02±1.23)％和(29.76±2.74)％,E-cadherin阳性率为(86.27±6.75)％,与荷瘤组比较差异有统计学意义(P＜0.05).LY294002组和淫羊藿素组组间比较差异无统计学意义(P＞0.05).结论 淫羊藿素具有调控P13K/Akt信号通路的作用,主要通过调控p-AR 、p-Akt、AR-V7和Calctionin水平而影响前列腺癌LNCaP细胞侵袭和转移.%Objective:To observe the effect of Epimedium on the signaling pathway of phosphatidylinositol 3-kinase(PI3K)/protein kinase B (Akt) and E-calcium mucin in the prostate cancer tissue of androgen dependent prostate cancer model mice,and to explore the mechanism.Methods:40 BALB/c-nu nude mice were randomly divided into the blank control group,tumor-burdened group,LY294002 group and epimedium group,10 cases in each group.Except the blank control group,the other groups were injected with 2*107/mL human LNCaP prostate cancer cell suspension in the prostate to establish the LNCaP tumor mouse model of prostate cancer.After two weeks,epimedium groups was injected with 80 mg/kg of epimedium by caudal vein for 4 weeks;in PI3K/Akt group,PI3K/Akt inhibitors (LY294002) were given tail-intravenously;in blank control group and tumor-burdened group,the same volume of physiological saline were injected tail-intravenously.After treatment,the prostate tissue was taken,and Western blotting was used to detect the phosphorylation AR (p-AR),phosphorylated protein kinase(p-Akt),and androgen receptor splicing variants 7 (AR-V7).The immunofluorescence double staining (FITC-CY3) method was used to detect the expressions of calcitonin and e-cadherin.Results:In tumor-burdened group,p-AR and p-Akt had high expressions in prostate tissue;AR-V7 and calcitonin positive rate were (32.45±2.15)％ and (46.31±5.01)％,respectively.E-cadherin positive rate was (22.93±1.96)％;compared with the blank control group,differences were statistically significant (P＜0.05).There was lower expression of p-AR and p-Akt in the prostate tissues of epihopin group;the positive rates of AR-V7 and Calctionin were (17.02±1.23) ％ and (29.76±2.74) ％,respectively;the E-cadherin positive rate was (86.27±6.75) ％;compared with the tumor-burdened group,differences were statistically significant (P ＜ 0.05).There was no statistically significant difference between the groups of LY294002 and icariin group(P＞ 0.05).Conclusion:Icariin plays a role in regulating the PI3K/Akt signaling pathway and affects the invasion and metastasis of prostate cancer LNCaP cells mainly by regulating the level of p-AR,p-Akt,AR-V7 and Calctionin.