目的 明确王不留行逆转人前列腺癌C4-2细胞激素非依赖性生长的单体成分，探究王不留行抑制去势抵抗性前列腺癌发生发展的作用机制。方法 采用CCK-8法检测王不留行单体成分对C4-2细胞增殖的影响；采用CCK-8法联合Annexin V-APC法检测王不留行单体成分对C4-2细胞雄激素非依赖性生长的作用；采用Western blotting检测磷脂酰肌醇3-激酶（phosphatidylinositol 3-kinase，PI3K）/蛋白激酶B（protein kinase B，Akt）通路和雄激素受体（androgen receptor，AR）蛋白表达；过表达AR后，采用荧光素酶报告基因检测AR的转录活性。结果 王不留行环肽B对C4-2细胞增殖的抑制作用最为显著。在有雄激素的条件下，王不留行环肽B对C4-2细胞凋亡的诱导作用和对细胞增殖的抑制作用较弱；王不留行环肽B组p-PI3K、p-Akt蛋白表达水平降低。过表达AR后，与对照组比较，双氢睾酮组AR蛋白表达水平和相对荧光素酶活力显著升高（P＜0.05），王不留行环肽B组AR表达水平和相对荧光素酶活力显著降低（P＜0.05）；与双氢睾酮组比较，双氢睾酮＋王不留行环肽B组AR蛋白表达水平和相对荧光素酶活力显著降低（P＜0.05）。结论 王不留行环肽B能明显抑制C4-2细胞增殖，具有逆转C4-2细胞激素非依赖性生长的作用，可以通过AR以及PI3K/Akt信号通路发挥抑制去势抵抗性前列腺癌发生发展的作用。

Objective To clarify the monomeric components of Vaccaria segetalis on reversing hormone independent growth of human prostate cancer C4-2 cells, and explore the mechanism of V. segetalis on inhibiting the occurrence and development of castration resistant prostate cancer. Methods CCK-8 method was used to detect the effect of monomeric components of V. segetalis on proliferation of C4-2 cells; CCK-8 method combined with Annexin V-APC method was used to detect the effect of monomeric components of V. segetalis on androgen independent growth of C4-2 cells; Western blotting was used to detect the expressions of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway and androgen receptor (AR) proteins; After overexpression of AR, the transcriptional activity of AR was detected using luciferase reporter genes. Results The inhibitory effect of segetalin B on the proliferation of C4-2 cells was the most significant. Under the conditions of androgens, the induction of apoptosis and inhibition of cell proliferation by segetalin B were relatively weak in C4-2 cells; The expression levels of p-PI3K and p-Akt proteins in segetalin B group were decreased. After overexpression of AR, compared with control group, the expression level of AR protein and relative luciferase activity in dihydrotestosterone group were significantly increased (P < 0.05), while the expression level of AR and relative luciferase activity in segetalin B group were significantly reduced (P < 0.05); Compared with dihydrotestosterone group, the expression level of AR protein and relative luciferase activity in dihydrotestosterone + segetalin B group were significantly reduced (P < 0.05). Conclusion Segetalin B can significantly inhibit the proliferation of C4-2 cells, reverse the hormone independent growth of C4-2 cells, and exert an inhibitory effect on the occurrence and development of castration resistant prostate cancer through AR and PI3K/Akt signaling pathway.

R285.5

天津中医药大学第一附属医院拓新工程（院YB202125）