连翘脂素调节JAK2/STAT3信号通路对前列腺癌增殖、凋亡和上皮间质转化的影响

Impacts of phillygenin on proliferation,apoptosis,and epithelial mesenchymal transformation in prostate cancer by regulating Janus kinase 2/signal transduction and transcription activating factor 3 signaling pathway

目的探讨连翘脂素(PHI)对前列腺癌(PCa)增殖、凋亡和上皮间质转化的影响及其机制。方法体外培养DU-145和LNCap-FGC细胞,将细胞分为对照组(Control组)、连翘脂素组(PHI组)、Janus激酶2(JAK2)抑制剂组(Ag490组)、连翘脂素+激活剂组(PHI+Colivelin组),分别检测细胞活力、细胞克隆能力、划痕愈合率及细胞侵袭;免疫组化法分析E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)和波形蛋白(Vimentin)表达;Westernblot检测JAK2、p-JAK2、信号转导和转录激活因子3(STAT3)、p-STAT3蛋白表达。结果与Control组比较,PHI组与Ag490组DU-145和LNCap-FGC胞存活率、细胞克隆数、划痕愈合率、细胞侵袭数、N-cadherin、Vimentin及p-JAK2、p-STAT3表达显著降低,细胞凋亡率和E-cadherin表达显著升高(P<0.05);PHI组与Ag490组比较,差异无统计学意义(P>0.05)。与PHI组比较,PHI+Colivelin组细胞存活率、细胞克隆数、细胞划痕愈合、细胞侵袭数、N-cadherin、Vimentin及p-JAK2、p-STAT3表达显著增加,细胞凋亡率和E-cadherin显著降低(P<0.05)。结论PHI可抑制JAK2/STAT3信号通路抑制PCa细胞增殖并诱导细胞凋亡,逆转DU-145和LNCap-FGC上皮间质转化进程。

Objective To investigate the impacts of phillygenin(PHI)on proliferation,apoptosis,and epithelial mesenchymal transformation in prostate cancer(PCa)and its mechanism.Methods DU-145 and LNCap-FGC cells were cultured in vitro and divided into control group,phillygenin group(PHI group),Janus kinase 2(JAK2)inhibitor group(Ag490 group),and phillygenin+activator group(PHI+Colivelin group).Cell viability,cell cloning ability,scratch healing rate,and cell invasion were detectede respectively.Immunohistochemistry was applied to analyze the expression of E-cadherin,N-cadherin and Vimentin.Western blot was applied to detect the expression of JAK2,p-JAK2,signal transduction and transcriptional activator 3(STAT3),p-STAT3 proteins.Results Compared with the control group,the cell survival rate,cell clone number,scratch healing rate,cell invasion number,N-cadherin,Vimentin,p-JAK2 and p-STAT3 expression of DU-145 and LNCap-FGC cells in the PHI group and Ag490 group were significantly decreased,the apoptosis rate and E-cadherin expression were significantly increased(P<0.05),and there was no significant difference between PHI group and Ag490 group(P>0.05).Compared with the PHI group,the cell survival rate,cell clone number,cell scratch healing,cell invasion number,N-cadherin,Vimentin,p-JAK2 and p-STAT3 expression in the PHI+Colivelin group were significantly increased,the apoptosis rate and E-cadherin were significantly decreased(P<0.05).Conclusions PHI can inhibit the JAK2/STAT3 signaling pathway to inhibit the proliferation and induce apoptosis of PCa cells,and reverse the process of epithelial mesenchymal transformation of DU-145 and LNCap-FGC.