结肠癌转移相关SMC1A基因剪切转录本之间的内源性竞争调控关系分析

It has been well concerned that SMC1A gene plays an important role in the metastasis initiating process of colon cancer. The alternative splicing of SMC1A produces six different transcript isoforms, two of which, named SMC1A-001 and SMC1A -201 can be translated into mature protein products.Tt is confirmed that SMC1A-001 can promote tumor metastasis, while SMC1A -201 lacks this ablity. Thus, further study is needed on the functional discrepancy among different SMC1A transcripts and the mechanism of endogenous regulation process. Our study will analyzes the differential expression of the six transcription isoforms. We will predict the functional miRNA using software and explore the characteristic of competitive binding process between miRNA and transcripts using RNA pull downand q-PCR test. Also, using CRISPR/CAS9 knocking out the exon2 of SMC1A-201,we have constructed a homozygous cell line that only produce the transcript SMC1A-001.And then, investigate the regulation effect that other transcripts adjust the expression of SMC1A and the related miRNA. It is of great significance for revealing how different transcripts of one gene could act as ce RNA and regulate the process of tumor metastasis, though clarifying the endogenous adjustment mechanism among transcripts and the related miRNA.

SMC1A基因在结肠癌转移启动过程中发挥重要功能。SMC1A基因选择性剪切产生6个转录本，只有SMC1A-001和SMC1A-201可以翻译出成熟蛋白产物。001的转移能力显著，201缺乏转移能力。SMC1A不同转录本的功能差异原因及其之间内源性调节机制有待深入分析。本研究分析SMC1A基因不同转录本的表达差异及其分别对SMC1A-001和SMC1A-201的调控关系，软件预测转录本之间的miRNA，RNA pull down和qPCR验证miRNA及竞争性结合特点。使用CRISPR/Cas9技术Knockout SMC1A-201 中 exon2，构建只产生001的pure-001 细胞株，再次分析转录本对SMC1A-001的调节关系及相关miRNA。研究转录本之间内源性调节的精细环节及发挥作用的miRNA，揭示转录本之间的内源性竞争的ceRNA作用，具有重要的意义。

SMC1A基因在结肠癌转移启动过程中发挥重要功能。SMC1A基因选择性剪切产生6个转录本 ，只有SMC1A-001和SMC1A-201可以翻译出成熟蛋白产物。001的转移能力显著，201缺乏转移能力。SMC1A不同转录本的功能差异原因及其之间内源性调节机制已做深入分析。本研究分析SMC1A基因不同转录本的表达差异及其分别对SMC1A-001和SMC1A-201的调控关系，软件预测转录本 之间的miRNA，RNA pull down和qPCR验证miRNA及竞争性结合特点。使用CRISPR/Cas9技术Knoc kout SMC1A-201 中 exon2，构建只产生001的pure-001 细胞株，再次分析转录本对SMC1A-001 的调节关系及相关miRNA。研究转录本之间内源性调节的精细环节及发挥作用的miRNA，揭示转录本之间的内源性竞争的ceRNA作用，具有重要的意义。初步探索SMC1A在结直肠肿瘤转移启动中的功能和作用机制，有助于进一步深入理解结直肠肿瘤分子生物学机制。本课题目前已接收及发表研究论著4篇，参加AACR举办的国际性肿瘤研究大会一次，壁报展示SMC1A研究成果；培养博士研究生3名，硕士研究生3名。