INSIG2促进宫颈腺癌恶性进展的分子机制研究

INSIG2 is a protein screened by proteomics technology, with highly expression and unknown function in cervical adenocarcinoma tissues. Bioinformatics analyses suggested that high expression of INSIG2 was associated with unfavorable overall survival in TCGA and GEO data. We previously found that the establishment of INSIG2 knockout cell lines via CRISPR/Cas9 mediated gene targeting could significantly reduce the proliferation, invasion and metastasis of cervical adenocarcinoma. The experimental result further showed that knockout of INSIG2 may influence TGF-β/Smad signaling pathway, promote epithelial-mesenchymal transition (EMT). Accordingly, we propose a hypothesis that INSIG2 regulates the malignant behavior of cervical adenocarcinoma by activating TGF-β/Smad signaling pathway. In this study, we will further systematically evaluate the function of IINSIG2 in vitro and in vivo by means of overexpression and CRISPR/Cas9 knockout technology. Finally, we investigated the molecular mechanism of INSIG2 in cervical adenocarcinoma via Pull-down、Co-IP and rescue experiments. If successful, this study may provide a potential target for the treatment of cervical adenocarcinoma.

INSIG2是我们采用蛋白质组学技术，在宫颈腺癌组织中筛选获得的、显著高表达于宫颈腺癌组织的蛋白。该蛋白在宫颈腺癌中尚未见报道。生物信息学分析提示，INSIG2在TCGA和GEO大样本数据库中表达水平越高，生存时间越短。前期实验发现，敲除INSIG2蛋白显著降低宫颈腺癌细胞的增殖、迁移和侵袭；进一步实验结果发现，敲除INSIG2蛋白可影响TGF-β/Smad信号通路，促进上皮间充质转化（EMT），故提出NSIG2可能通过调控TGF-β/Smad信号通路，导致EMT发生，促进宫颈腺癌恶性进展的假说。本研究拟采用过表达与CRISPR/Cas9技术，通过体外和体内两方面系统评估INSIG2对宫颈腺癌恶性进展的影响；采用Pull-down、Co-IP以及挽救策略，充分论证INSIG2在宫颈腺癌中的分子机制。本研究如获成功，将有望为宫颈腺癌的治疗提供新的靶标。

宫颈腺癌在过去的几十年中发病率升高，因为其检出率低，恶性行为高。胰岛素诱导基因2(INSIG2)，一种膜蛋白内质网(ER)在癌症进展中起着至关重要的作用INSIG2和宫颈腺癌之间的关系鲜为人知。应用人类蛋白质图谱(HPA)和癌症基因组图谱(TCGA)宫颈癌(CESC)数据来研究INSIG2表达的改变.进行生物学功能检测恶性程度的变化行为。生物信息学分析以探索 INSIG2在宫颈腺癌进展中的潜在影响。我们的研究证实INSIG2高表达的患者预后较差，敲低INSIG2可以抑制宫颈腺癌细胞的增殖、迁移、侵袭，并下调EMT相关基因表达水平.基因富集分析显示了很多差异表达基因,揭示了宫颈腺癌进展中INSIG2更多的潜在功能。