The regulating effect of Eg5 on the malignant proliferation of castration resistant prostate cancer cells through c-Myc/SP1/CDK1 signaling pathway
HU Junbiao1, ZHANG Chunting1, XIA Jianhong2, YAO Yong1,LU Junyi1
1.Department of Urinary Surgery, People’s Hospital of Jinhua, Jinhua 321000, China; 2.Department of Pharmacy, People’s Hospital of Jinhua, Jinhua 321000, China
HU Junbiao,ZHANG Chunting,XIA Jianhong, et al. The regulating effect of Eg5 on the malignant proliferation of castration resistant prostate cancer cells through c-Myc/SP1/CDK1 signaling pathway[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2021, 51(9): 705-712.
Abstract：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-cadherin expression 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.
