Primary cilia, regulated via distinct signal transduction pathways, play crucial roles in various cellular behaviors. However, the full regulatory mechanism involved in primary cilia development during cellular differentiation is not fully understood, particularly for the sensory hair cells of the mammalian cochlea. In this study, we investigated the effects of the Rho-kinase inhibitor Y27632 and PKCα inhibitor GF109203X on primary cilia-related cell behavior in undifferentiated and differentiated temperature-sensitive mouse cochlear precursor hair cells (the conditionally immortalized US/VOT-E36 cell line). Our results indicate that treatment with Y27632 or GF109203X induced primary cilia elongation and tubulin acetylation in both differentiated and undifferentiated cells. Concomitant with cilia elongation, Y27632 treatment also increased Hook2 and cyclinD1 expression, while only Hook2 expression was increased after treatment with GF109203X. In the undifferentiated cells, we observed an increase in the number of S and G2/M stage cells and a decrease of G1 cells after treatment with Y27632, while the opposite was observed after treatment with GF109203X. Finally, while both treatments decreased oxidative stress, only treatment with Y27632, not GF109203X, induced cell cycle-dependent cell proliferation and cell migration.

中文翻译：

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Rho激酶和PKCα抑制作用诱导原发纤毛伸长并改变未分化和分化的温度敏感小鼠耳蜗细胞的行为。

通过不同的信号转导途径调节的初级纤毛在各种细胞行为中起着至关重要的作用。但是，尚未完全了解在细胞分化过程中初级纤毛发育所涉及的完整调节机制，特别是对于哺乳动物耳蜗的感觉毛细胞。在这项研究中，我们研究了Rho激酶抑制剂Y27632和PKCα抑制剂GF109203X对未分化和分化的温度敏感小鼠耳蜗前体毛细胞（条件无限增殖的US / VOT-E36细胞系）中初级纤毛相关细胞行为的影响。 。我们的结果表明，在分化和未分化细胞中，用Y27632或GF109203X进行处理均可诱导原发性纤毛伸长和微管蛋白乙酰化。伴随着纤毛伸长，Y27632处理还增加了Hook2和cyclinD1的表达，而仅GF109203X处理后Hook2的表达增加了。在未分化的细胞中，用Y27632处理后，观察到S和G2 / M期细胞数量增加，而G1细胞减少，而用GF109203X处理后，观察到相反的情况。最后，尽管两种处理均降低了氧化应激，但仅使用Y27632而非GF109203X进行处理即可诱导细胞周期依赖性细胞增殖和细胞迁移。