Min Sha-Sha, Cai Zhong-Lin, Shen Yan-Ting, Wang Zhong
School of Gongli Hospital Medical Technology， University of Shanghai for Science and Technology， Shanghai 200093， China.
Department of Urology， Shanghai Pudong New Area Gongli Hospital, Shanghai 200135， China.
Zhonghua Nan Ke Xue. 2025 Sep;31(9):791-799.
To investigate the expression of RNA binding protein ELAVL1 in prostate cancer (PCa), especially hormone-sensitive prostate cancer (HSPC), and its relationship with tumor proliferation. This study further aims to reveal the molecular mechanism by which ELAVL1 promotes HSPC proliferation by stabilizing SOX4 mRNA in an m6A-dependent manner.
The expression of ELAVL1 in PCa tissues and its relationship with prognosis were analyzed in the Cancer Genome Atlas (TCGA) database, and the differences in HSPC and hormone-resistant prostate cancer (HRPC) were compared. And its relationship with prognosis were analyzed in the Cancer Genome Atlas (TCGA) database, and the differences in HSPC and hormone-resistant prostate cancer (HRPC) were compared. Western blot was used to detect ELAVL1 protein expression in PCa cell lines. After ELAVL1 knockdown by siRNA, cell proliferation was evaluated using CCK-8 assays, and changes in downstream target genes were detected by RT-qPCR. Tumor xenograft experiments in nude mice were performed to further assess the impact of ELAVL1 on tumor growth. The interaction between ELAVL1 and SOX4 mRNA was verified by RIP-seq. And the mRNA and protein levels of SOX4 after knockdown of ELAVL1 were detected by RT-qPCR and Western blot, respectively. CCK-8 was used to evaluate the effect of SOX4 knockdown on cell proliferation. MeRIP-qPCR was used to detect the m6A modification level of SOX4 and the effect of knocking down METTL3. RNA pull-down experiments verified the interaction between SOX4 RNA fragments and ELAVL1 protein. RNA stability experiments evaluated the effect of ELAVL1 knockdown on SOX4 mRNA stability.
The expression of ELAVL1 in PCa cells was higher than that in normal prostate epithelial cells. The prognosis of patients with high expression of ELAVL1 was significantly worse than that of patients with low expression. In the GSE32269 dataset, the expression level of ELAVL1 in HSPC was significantly higher than that in HRPC. After knocking down of ELAVL1 in LNCaP and VCaP cells, CCK-8 experiments showed that the cell proliferation ability was significantly affected after knocking down ELAVL1, and overexpressed ELAVL1 promoted the proliferation of HSPC cells. The results of in vivo studies showed that knockdown of ELAVL1 significantly inhibited the tumorigenic capacity of LNCaP cells and resulted in a marked reduction in xenograft tumor mass. The levels of SOX4 mRNA and protein in LNCaP and VCaP cells were significantly higher than those in normal prostate epithelial cells RWPE-1. RIP-qPCR confirmed the interaction between ELAVL1 protein and SOX4 mRNA. After knocking down of ELAVL1, the expression levels of SOX4 mRNA and protein were significantly decreased. After knocking down of SOX4, the proliferation ability of LNCaP and VCaP cells was significantly inhibited.
ELAVL1 is highly expressed in HSPC. High expression of ELAVL1 is associated with the proliferation of HSPC. SOX4 is a downstream molecule of ELAVL1 which promotes the proliferation of HSPC. ELAVL1 enhances the stability of SOX4 mRNA through an m6A-dependent mechanism.
研究RNA结合蛋白ELAVL1在前列腺癌（PCa）尤其是激素敏感性前列腺癌（HSPC）中的表达及其与肿瘤增殖的关系。本研究还旨在揭示ELAVL1通过m6A依赖性方式稳定SOX4 mRNA促进HSPC增殖的分子机制。
在癌症基因组图谱（TCGA）数据库中分析ELAVL1在PCa组织中的表达及其与预后的关系，并比较HSPC和激素抵抗性前列腺癌（HRPC）之间的差异。采用蛋白质免疫印迹法检测PCa细胞系中ELAVL1蛋白的表达。用小干扰RNA（siRNA）敲低ELAVL1后，采用CCK-8法评估细胞增殖情况，并用逆转录定量聚合酶链反应（RT-qPCR）检测下游靶基因的变化。进行裸鼠肿瘤异种移植实验，进一步评估ELAVL1对肿瘤生长的影响。通过RNA免疫沉淀测序（RIP-seq）验证ELAVL1与SOX4 mRNA之间的相互作用。分别用RT-qPCR和蛋白质免疫印迹法检测敲低ELAVL1后SOX4的mRNA和蛋白水平。用CCK-8法评估敲低SOX4对细胞增殖的影响。用甲基化RNA免疫沉淀定量PCR（MeRIP-qPCR）检测SOX4的m6A修饰水平及敲低甲基转移酶样蛋白3（METTL3）的效果。RNA下拉实验验证SOX4 RNA片段与ELAVL1蛋白之间的相互作用。RNA稳定性实验评估敲低ELAVL1对SOX4 mRNA稳定性的影响。
PCa细胞中ELAVL1的表达高于正常前列腺上皮细胞。ELAVL1高表达患者预后明显差于低表达患者。在GSE32269数据集中，HSPC中ELAVL1的表达水平明显高于HRPC。在LNCaP和VCaP细胞中敲低ELAVL1后，CCK-8实验表明敲低ELAVL1后细胞增殖能力明显受影响，而过表达ELAVL1可促进HSPC细胞增殖。体内研究结果表明，敲低ELAVL1显著抑制LNCaP细胞的致瘤能力，并导致异种移植瘤质量显著降低。LNCaP和VCaP细胞中SOX4的mRNA和蛋白水平明显高于正常前列腺上皮细胞RWPE-1。RIP-qPCR证实ELAVL1蛋白与SOX4 mRNA之间存在相互作用。敲低ELAVL1后，SOX4的mRNA和蛋白表达水平明显降低。敲低SOX4后LNCaP和VCaP细胞的增殖能力明显受到抑制。
ELAVL1在HSPC中高表达。ELAVL1高表达与HSPC增殖相关。SOX4是ELAVL1促进HSPC增殖的下游分子。ELAVL1通过m6A依赖性机制增强SOX4 mRNA的稳定性。
