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为了研究前列腺癌相关基因(prostate and colon gene 1,PC-1)对受体酪氨酸激酶家族分子EphA3表达的影响,用RT-PCR、实时PCR和Western印迹检测表达不同水平PC-1的前列腺癌细胞系LNCaP和C4-2中EphA3的表达情况.发现PC-1可诱导EphA3基因表达上调.采用荧光素酶实验检测PC-1对于EphA3启动子转录活性的影响,结果显示,PC-1对转录起始位点上游916 bp的启动子活性没有影响,而可增强转录起始位点上游2011 bp启动子的活性.对EphA3启动子-916 bp～-2 011 bp区域进行生物信息学分析,结果显示,此区域包含HSF、NF-1、Nkx-2、SP1和GATA-1等多种转录因子结合位点.实验结果表明,PC-1可通过影响EphA3启动子诱导EphA3基因高表达,其调控区域位于转录起始位点上游-2 011 bp至-916 bp之间,提示PC-1可能通过影响一些结合于此区域的转录因子来影响EphA3启动子的转录活性.
In order to investigate the effect of prostate and colon gene 1 (PC-1) on receptor tyrosine kinase family molecule EphA3 expression, prostate-PC-1-expressing prostate was tested by RT-PCR, real-time PCR and Western blotting The expression of EphA3 in cancer cell lines LNCaP and C4-2 was found to be upregulated by PC-1.PCR assay was used to detect the effect of PC-1 on the transcriptional activity of EphA3 promoter. The results showed that PC- The 916 bp promoter upstream of the transcriptional start site had no effect on the promoter activity of the 2011 bp promoter upstream of the transcription start site.Bioinformatics analysis was performed on the -916 bp ~ -2 011 bp region of the EphA3 promoter, The results showed that this region contains binding sites of many transcription factors such as HSF, NF-1, Nkx-2, SP1 and GATA-1.The experimental results show that PC-1 can induce EphA3 gene overexpression by affecting EphA3 promoter, The regulatory region is located between -2 011 bp and -916 bp upstream of the transcriptional start site, suggesting that PC-1 may affect the transcriptional activity of the EphA3 promoter by affecting some transcription factors that bind to this region.