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作者	单位	E-mail
李彤	复旦大学附属中山医院肝肿瘤内科上海 200032；复旦大学附属中山医院徐汇医院消化内科上海 200031	li.tong@zs-hospital.sh.cn
郭婧	复旦大学附属中山医院徐汇医院消化内科上海 200031
李东	上海嘉会国际医院普通外科上海 200233
中山静子	复旦大学附属中山医院消化内科上海 200032
余湘南	复旦大学附属中山医院消化内科上海 200032
徐馨	复旦大学附属中山医院肝肿瘤内科上海 200032

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中文摘要:

摘要目的：探讨SKA1调控胰腺导管腺癌（PDAC）侵袭与转移的分子机制。方法：利用免疫组化染色法检测胰腺导管腺癌组织样本及癌旁组织中SKA1与下游分子Cdc42的表达水平、用免疫印迹方法在不同胰腺癌细胞系及正常胰腺导管上皮细胞中进行验证；利用慢病毒转染技术构建SKA1稳定敲减和过表达的胰腺癌细胞株；利用免疫荧光方法检测PDAC细胞内SKA1敲低或过表达对高尔基体结构的影响，及Cdc42抑制剂ZCL278对高尔基体结构变化的调控；并利用Transwell实验及细胞划痕实验检测ZCL278对SKA1促癌作用的影响；应用免疫印迹方法检测SKA1及Cdc42表达对自噬标志物的影响。结果：在胰腺导管腺癌组织与细胞中SKA1与Cdc42表达均显著高于正常组织或细胞，且二者表达呈显著正相关；并且发现SKA1低表达的患者具有更长的总体生存期，而Cdc42表达高低与总体生存期无关。接着我们发现SKA1可促进PDAC细胞内高尔基体堆叠，并且在SKA1过表达的细胞中Cdc42抑制剂ZCL278可以抑制这种堆叠现象。进一步我们发现抑制Cdc42可逆转SKA1过表达对胰腺癌侵袭与转移的促进效应，具有统计学差异，而在SKA1非高表达的细胞中无显著差异，最后我们发现Cdc42可受SKA1的表达调控诱导PDAC细胞发生自噬。结论：SKA1通过调控Cdc42介导的高尔基体致密堆叠进而影响胰腺癌发生自噬，最终促使胰腺癌细胞迁移和侵袭。

英文摘要:

ABSTRACT Objective: To explore the molecular mechanism of SKA1 regulating invasion and metastasis of pancreatic ductal adenocarcinoma (PDAC). Methods: Immunohistochemical staining was used to detect the expression levels of SKA1 and Cdc42 in pancreatic cancer tissue samples and paired para-cancer tissues, and western blot was used to verify the expression levels of SKA1 and Cdc42 in different pancreatic cancer cell lines and normal pancreatic ductal epithelial cells. Pancreatic cancer cell lines with SKA1 stable knockdown and overexpression were constructed by lentivirus transfection technique. The effect of SKA1 knockdown or overexpression on Golgi structure in PDAC cells and the regulation of Cdc42 inhibitor ZCL278 on Golgi structure were detected by immunofluorescence method. The effect of ZCL278 on cancer metastasis induced by SKA1 overexpression was detected by Transwell assay and cell scratch formation assay. The effects of SKA1 and Cdc42 expression on autophagy markers were detected by western blotting. Results: The expressions of SKA1 and Cdc42 in pancreatic ductal adenocarcinoma tissues and cells were significantly higher than those in normal tissues or cells, and the expressions of SKA1 and CDC42 were significantly positively correlated. Patients with low SKA1 expression were found to have a longer overall survival time, while high or low Cdc42 expression was not associated with overall survival. We then found that SKA1 promotes Golgi stacking in PDAC cells and that the Cdc42 inhibitor ZCL278 inhibits this stacking phenomenon in SKA1 overexpressed PDAC cells. Further, we found that inhibition of Cdc42 could reverse the promoting effect of SKA1 overexpression on pancreatic cancer invasion and metastasis, with statistical difference, while there was no significant difference in cells with non-high SKA1 expression. Finally, we found that Cdc42 could induce autophagy of PDAC cells by the regulation of SKA1. Conclusion: SKA1 influences autophagy of PDAC by regulating the dense Golgi stacking mediated by Cdc42, and finally promotes the migration and invasion of pancreatic cancer cells.

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