UGT8对结直肠癌生长迁移的影响及与SOX9相关性研究

Effect of UGT8 on colorectal cancer cell proliferation and migration and its correlationwith SOX9 expression

    摘要
  • 摘要:
    目的 探讨尿苷二磷酸半乳糖转移酶（UDP galactosyltransferase 8，UGT8）对结直肠癌细胞生长迁移的影响及可能的机制，以及性别决定区 Y 框蛋白 9（SRY-box transcription factor 9，SOX9）对UGT8可能的调控作用。
    方法 基于GEPIA2、UALCAN和TIMER 2.0在线数据库分析结直肠癌组织中UGT8和SOX9的表达水平及两者表达水平的相关性。通过免疫组织化学法检测结直肠癌及其癌旁组织的UGT8和SOX9的蛋白表达情况并分析其与临床病理特征的关系及两者表达水平相关性；在结直肠癌细胞中沉默UGT8后，CCK-8实验检测细胞的增殖能力；Transwell和划痕实验检测细胞的迁移能力；Western blot实验检测EMT标志物（E-cadherin和ZEB1）蛋白表达情况；RT-qPCR和Western blot法检测沉默SOX9后UGT8的mRNA和蛋白表达情况；JASPER在线网站预测SOX9是否与UGT8的启动子结合。
    结果 生物信息学结果显示，与正常组织相比较，UGT8和SOX9的mRNA表达水平异常升高，而且两者表达水平呈正相关。免疫组织化学结果显示，UGT8、SOX9在结直肠癌组织中的表达水平明显高于癌旁组织；UGT8的蛋白表达水平与N分期相关；SOX9的表达水平与T分期相关；UGT8和SOX9在结直肠癌组织中的表达存在正相关。沉默UGT8后细胞的增殖能力减弱；细胞的迁移能力降低；E-cadherin蛋白表达增加，ZEB1蛋白表达降低。沉默SOX9后，UGT8的mRNA和蛋白水平显著降低；JASPER网站预测SOX9可以和UGT8的启动子特异序列结合。
    结论 UGT8和SOX9在结直肠癌病理组织中均高表达，与临床病理特征相关，而且两者表达水平呈正相关；UGT8通过促进上皮间质转化促进结直肠癌细胞的增殖和迁移；SOX9促进UGT8的mRNA和蛋白表达，并可能与UGT8启动子结合。

     

    Abstract:
    Objective To investigate the effect of uridine diphosphate ceramide galactosyltransferase 8 (UGT8) on colorectal cancer (CRC) cell growth and migration, elucidate an underlying mechanism, and assess the potential regulatory role of SRY-box transcription factor 9 (SOX9) on UGT8.
    Methods UGT8 and SOX9 mRNA expression levels in CRC tissues, and correlation between their expression levels, were analyzed using GEPIA2, UALCAN, and TIMER 2.0 online databases. UGT8 and SOX9 protein expression in CRC and adjacent tissues was detected using immunohistochemistry, and relationships between their expression and clinicopathological characteristics were analyzed. Impact of UGT8 knockdown on CRC cell proliferation was assessed using a CCK-8 assay, and cell migration was evaluated using Transwell and wound healing assays. Western blot was performed to detect expression of epithelial-mesenchymal transition (EMT) markers (E-cadherin and ZEB1). RT-qPCR and Western blot were used to measure UGT8 mRNA and protein expression levels after SOX9 knockdown. The JASPAR online database was used to assess SOX9 potential for binding to the UGT8 promoter.
    Results Bioinformatics analyses revealed significantly higher mRNA expression levels of both UGT8 and SOX9 in CRC tissues than in normal tissues. Positive correlation was observed between expression levels. Immunohistochemistry results showed that tumor UGT8 and SOX9 protein levels were significantly higher than those in adjacent tissues. UGT8 protein level was found to correlates with N stage, and SOX9 protein level correlated with T stage. A positive correlation was observed between UGT8 and SOX9 expression levels. Following UGT8 knockdown, cell proliferation capacity was attenuated and cell migration ability was reduced. E-cadherin expression concurrently increased and ZEB1 expression decreased. RT-qPCR and Western blot results showed that SOX9 knockdown significantly reduced UGT8 mRNA and protein levels. The JASPER website predicts that SOX9 will bind to the UGT8 promoter.
    Conclusions UGT8 and SOX9 are highly expressed in CRC tissues, and their expression levels correlate with clinicopathological features. UGT8 and SOX9 expression levels display significant positive correlation. Mechanistically, UGT8 promotes CRC cell proliferation and migration by facilitating epithelial-mesenchymal transition (EMT). SOX9 enhances UGT8 mRNA and protein expression and may bind to the UGT8 promoter region.

     

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