ASF1B通过调控P53相关信号通路促进前列腺癌迁移和增殖的研究

ASF1B enhances the migration and proliferation of prostate cancers via regulating p53-related signaling pathways

目的研究组蛋白伴侣抗沉默功能1B(ASF1B)对前列腺癌(PCa)迁移和增殖能力的影响,探索ASF1B调控P53相关信号通路的具体分子机制。方法根据TCGA数据库中的数据分析ASF1B在前列腺癌患者中的生存预后情况,使用细胞小干扰RNA(SiRNA)敲低ASF1B表达后,通过Transwell细胞迁移实验比较细胞的运动能力,通过CCK8实验和平板克隆实验比较细胞的增殖能力,使用细胞凋亡实验比较细胞的凋亡率,通过转录组二代测序比较敲低ASF1B后各个信号通路的改变情况,通过Western-Blot实验及Q-PCR实验比较P53相关信号通路分子的蛋白质和mRNA水平变化,以阐明ASF1B对前列腺癌细胞的迁移及增殖能力的影响。结果SiRNA敲低ASF1B表达后,DU145和PC3细胞的运动能力降低,相同时间内穿过小室的细胞数减少。敲低ASF1B表达后,DU145和PC3细胞的细胞活力和集落形成个数及大小降低,细胞增殖能力下降。敲低ASF1B表达后,DU145和PC3细胞的凋亡率增加。敲低ASF1B表达后,PC3细胞的转录组二代测序结果提示P53信号通路受到调控(P<0.01)。敲低ASF1B表达后,PC3细胞内P53相关通路的P21、IGFBP3、BBC3表达量上升,Cyclin D、Snail、Slug表达量下降。结论ASF1B以非P53依赖的形式通过P53相关信号通路调控前列腺癌细胞的迁移及增殖,ASF1B有望成为前列腺癌的诊断及治疗的新型靶点。

Objective To study the effect of Anti-Silencing Function 1B Histone Chaperone(ASF1B)on the migration and proliferation of prostate cancer(PCa),and to explore the specific molecular mechanism of ASF1B regulating P53-related signal pathway.Methods The survival and prognosis of ASF1B in prostate cancer patients was analyzed according to the data in TCGA database.After the expression of ASF1B was knocked down by small interference RNA(SiRNA),the mobility of cells was compared by Transwell cell migration test,the ability of cell proliferation was compared by CCK8 test and plate cloning test,and the apoptosis rate was compared by apoptosis test.The changes of signal pathways after ASF1B knockout were compared by transcriptome second generation sequencing,and the changes of protein and mRNA levels of P53 related signal pathway molecules were compared by Western-Blot test and Q-PCR test,in order to clarify the effect of ASF1B on the migration and proliferation of prostate cancer cells.Results After SiRNA knocked down the expression of ASF1B,the motor ability of DU145 and PC3 cells decreased,and the number of cells passing through the chamber decreased at the same time.After knocking down the expression of ASF1B,the number and size of cell viability and colony formation of DU145 and PC3 cells decreased,and the ability of cell proliferation decreased.After knocking down the expression of ASF1B,the percentage of apoptosis of DU145 and PC3 cells increased.After knocking down the expression of ASF1B,the second generation sequencing results of transcriptional group of PC3 cells showed that the P53 signal pathway was regulated.After knocking down the expression of ASF1B,the expression of p21,IGFBP3 and BBC3 of P53-related pathways in PC3 cells increased,while the expression of Cyclin D,Snail and Slug decreased.Conclusion ASF1B regulates the migration and proliferation of prostate cancer cells through P53-related signaling pathways in a P53-independent manner.ASF1B is expected to be a new target for the diagnosis and treatment of prostate cancer.