Tianjin Institute of Urology, The Second Hospital of Tianjin Medical University, Tianjin, China.
J Transl Med. 2023 Apr 26;21(1):281. doi: 10.1186/s12967-023-04118-2.
The primary cilia (PC) is a microtubule-based and nonmotile organelle which protrudes from the surface of almost all mammalian cells. At present, PC has been found to be a deficiency or loss in multiple cancers. Restoring PC could be a novel targeting therapy strategy. Our research showed that PC was reduced in human bladder cancer (BLCA) cells, and PC deficiency promotes cell proliferation. However, the concrete mechanisms remain unknown. SCL/TAL1 interrupting locus (STIL), a PC-related protein, was screened in our previous study and could influence the cell cycle by regulating PC in tumor cells. In this study, we aimed to elucidate the function of STIL for PC to explore the underlying mechanism of PC in BLCA.
Public database analysis, western blot, and enzyme-linked immunosorbent assay (ELISA) were used to screen genes and explore gene expression alteration. Immunofluorescence and western blot were utilized to investigate PC. Wound healing assay, clone formation assay, and CCK-8 assay were used to explore cell migration, growth, and proliferation. The co-immunoprecipitation and western blot were employed to reveal the interaction of STIL and AURKA.
We found that high STIL expression is correlated with poor outcomes of BLCA patients. Further analysis revealed that STIL overexpression could inhibit PC formation, activate SHH signaling pathways, and promote cell proliferation. In contrast, STIL-knockdown could promote PC formation, inactivate SHH signaling, and inhibit cell proliferation. Furthermore, we found that the regulatory functions of STIL for PC depend on AURKA. STIL could influence proteasome activity and maintain AURKA stabilization. AURKA-knockdown could reverse PC deficiency caused by STIL overexpression for PC in BLCA cells. We observed that co-knockdown in STIL and AURKA significantly enhanced PC assembly.
In summary, our result provides a potential therapy target for BLCA based on the restoration of PC.
原发性纤毛（PC）是一种由微管组成的非运动细胞器，从几乎所有哺乳动物细胞的表面伸出。目前，已经发现 PC 在多种癌症中存在缺陷或缺失。恢复 PC 可能是一种新的靶向治疗策略。我们的研究表明，PC 在人膀胱癌（BLCA）细胞中减少，PC 缺陷促进细胞增殖。然而，具体机制尚不清楚。在我们之前的研究中筛选出了与 PC 相关的蛋白 SCL/TAL1 中断位点（STIL），它可以通过调节肿瘤细胞中的 PC 来影响细胞周期。在这项研究中，我们旨在阐明 STIL 对 PC 的功能，以探讨 PC 在 BLCA 中的潜在机制。
使用公共数据库分析、western blot 和酶联免疫吸附试验（ELISA）筛选基因并探索基因表达变化。免疫荧光和 western blot 用于研究 PC。划痕愈合试验、克隆形成试验和 CCK-8 试验用于研究细胞迁移、生长和增殖。共免疫沉淀和 western blot 用于揭示 STIL 和 AURKA 的相互作用。
我们发现高 STIL 表达与 BLCA 患者的不良预后相关。进一步分析表明，STIL 过表达可抑制 PC 形成，激活 SHH 信号通路，并促进细胞增殖。相反，STIL 敲低可促进 PC 形成，使 SHH 信号失活，抑制细胞增殖。此外，我们发现 STIL 对 PC 的调节功能取决于 AURKA。STIL 可以影响蛋白酶体活性并维持 AURKA 的稳定性。AURKA 敲低可逆转 STIL 过表达引起的 BLCA 细胞中 PC 缺陷。我们观察到 STIL 和 AURKA 的共敲低显著增强了 PC 组装。
综上所述，我们的结果为 BLCA 基于 PC 恢复的治疗提供了一个潜在的靶点。
