肿瘤相关因子CD9在鹿茸干细胞中的表达检测及功能分析

发布时间：2018-09-10 18:06

【摘要】：CD9是一个肿瘤相关因子,早期的研究证实CD9的表达能够抑制肿瘤的生长和转移,后来有研究人员提出CD9表达量在肿瘤细胞中升高,并证实其表达促进肿瘤的发展,因此CD9在肿瘤发展过程中的角色仍然存在巨大争议。鹿茸再生过程中,细胞增殖速度甚至超过肿瘤,CD9同样在鹿茸干细胞中高表达。为了探讨鹿茸干细胞中CD9的功能,本研究利用Western blot,免疫荧光和流式细胞仪分析技术系统地检测了鹿茸干细胞(包括生茸区骨膜细胞(antlerogenic periosteum cells,APC),角柄骨膜细胞(pedicle periosteum cells,PPC)和鹿茸尖部间充质细胞(reserve mesenchymal cells,RMC))中CD9的表达情况;并利用Anti-CD9抗体抑制CD9活性,检测了CD9对鹿茸干细胞迁移和增殖的影响。结果显示,3种鹿茸干细胞均高水平表达CD9,原代培养的鹿茸干细胞CD9阳性率超过90%;抑制CD9活性,对鹿茸干细胞的迁移影响不显著,但对部分鹿茸干细胞(PPC和鹿茸RMC)的增殖有促进作用;分析了CD9有关的细胞基质-整合素和细胞粘附信号通路,并从鹿茸干细胞转录组中检测到绝大多数因子。本研究揭示了CD9在鹿茸干细胞中的功能并推测了可能的调控机制,将有助于揭示CD9在快速增殖的细胞特别是肿瘤细胞中的作用。
[Abstract]:CD9 is a tumor-related factor. Early studies have shown that the expression of CD9 can inhibit the growth and metastasis of tumor. Later, some researchers suggested that the expression of CD9 increased in tumor cells and promoted the development of tumor. Therefore, the role of CD9 in tumor development is still controversial. In the process of pilose antler regeneration, the cell proliferation rate was even higher than that of tumor CD9 expression in pilose antler stem cells. In order to investigate the function of CD9 in pilose antler stem cells, In this study, Western blot, immunofluorescence and flow cytometry were used to detect the expression of CD9 in pilose antler stem cells (including (antlerogenic periosteum cells,APC, (pedicle periosteum cells,PPC and (reserve mesenchymal cells,RMC). The effect of CD9 on migration and proliferation of pilose antler stem cells was detected by using Anti-CD9 antibody to inhibit CD9 activity. The results showed that the positive rate of CD9 of the three kinds of pilose antler stem cells expressing CD9, was higher than 90%, and the inhibition of CD9 activity had no significant effect on the migration of pilose antler stem cells. But it can promote the proliferation of some pilose antler stem cells (PPC and RMC), analyze the cell matrix integrin and cell adhesion signaling pathway related to CD9, and detect most of the factors from the transcription group of pilose antler stem cells. This study revealed the function of CD9 in pilose antler stem cells and speculated the possible regulatory mechanism, which will help to reveal the role of CD9 in rapidly proliferating cells, especially in tumor cells.
【作者单位】：中国农业科学院特产研究所/特种经济动物分子生物学国家重点实验室;
【基金】：国家自然科学基金项目(No.31500792) 吉林省自然科学基金(No.20170101032JC) 中国农业科学院科技创新工程项目(No.CAAS-ASTIP-2014-ISAPS-04)
【分类号】：S825

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