Eg5通过c-Myc/SP1/CDK1通路调节去势抵抗性前列腺癌细胞的恶性增殖

目的：探究纺锤体有丝分裂驱动蛋白（Eg5）对去势抵抗性前列腺癌细胞株DU145生长、凋亡、细胞周期及侵袭转移的调节作用及机制。方法：Eg5 shRNA干扰Eg5在DU145细胞内的表达，小分子抑制剂SB-743921抑制Eg5在DU145细胞内的功能。实验分为对照组、NC shRNA组、Eg5 shRNA组和SB-743921组，MTT实验和单克隆形成实验测定各组细胞生长；Annexin V-FITC和PI双染实验测定各组细胞凋亡；PI单染实验分析各组细胞周期；划痕实验分析各组细胞迁移能力；q-PCR和Western blot评价各组细胞内c-Myc、SP1、CDK1、Cyclin A1、Cyclin B1、CDC25B、N-cadherin、Vimentin及E-cadherin的表达情况。结果：NC shRNA组与对照组各指标差异均无统计学意义（P ＞0.05）。Eg5 shRNA组、SB-743921组与对照组比，细胞增殖及迁移能力降低、细胞凋亡增加，细胞周期阻滞在G2/M期，差异均有统计学意义（P ＜0.05）。Eg5 shRNA组、SB-743921组与对照组相比，细胞内c-Myc和SP1的表达量降低（P ＜0.05），调控G2/M期基因CDK1、Cyclin A1、Cyclin B1及CDC25B的表达降低（P ＜0.05），同时调控细胞迁移基因N-cadherin和Vimentin的表达量降低，E-cadherin的表达量增加（P ＜0.05）。结论：有丝分裂驱动蛋白Eg5可能通过c-Myc/SP1/CDK1通路调节去势抵抗性前列腺癌的恶性增殖和侵袭转移，是治疗前列腺癌的潜在靶点。

The regulating effect of Eg5 on the malignant proliferation of castration resistant prostate cancer cells through c-Myc/SP1/CDK1 signaling pathway

Objective: To investigate the effect of kinesin-5 (Eg5) on the proliferation, apoptosis, cell cycle,invasion and metastasis of castration resistant prostate cancer DU145 cells and its corresponding mechanism.Methods: Eg5 expression was silenced through transfecting DU145 cells with Eg5 shRNA and its function in DU145 cells was inhibited with SB-743921. The experiment were divided as the control group, NC shRNA group,Eg5 shRNA group and SB-743921 treated group. MTT assay and Colony formation assay were used to analyze cell proliferation. Annexin V-FITC and PI double dyeing assay was used to determine cell apoptosis and PI dyeing assay to detect cell cycle. Wound healing assay was used to analyze cell metastasis. Q-PCR and Western blot was used to analyze the expression of c-Myc, SP1, CDK1, Cyclin A1, Cyclin B1, CDC25B, N-cadherin, E-cadherin and Vimentin. Results: There was no significant difference between NC shRNA group and the control group.Compared with the control group, the proliferation and metastasis of DU145 cells were decreased in Eg5 shRNA group and SB-743921 treated group, while the apoptosis of DU145 cells was increased and the cell cycle was blocked at G2/M phase, with statistical difference among groups (P<0.05). The expression of c-Myc (P<0.05) and SP1 (P<0.05) were decreased in Eg5 shRNA group and SB-743921 treated group and the expression of G2/M phase related genes CDK1, Cyclin A1, Cyclin B1 and CDC25B were also inhibited (P<0.05). Meanwhile, the expression of metastasis related genes Vimentin and N-cadherin were blocked (P<0.05), and E-cadherinexpression was increased (P<0.05). Conclusion: As a potential therapeutic target, Eg5 may regulate the c-Myc/SP1/CDK1 signaling pathway to inhibit the proliferation and metastasis of Castration resistant prostate cancer.