摘要
[目的]PDCD6是一种程序性细胞死亡基因。近年来,已有不少研究证明PDCD6在一定程度上可以促进卵巢癌细胞的增殖和迁移。然而,它的确切分子机制还有待进一步验证。[方法]利用慢病毒sh RNA敲除HO-8910PM的PDCD6水平,以空载体作为阴性对照。提取两组细胞的总RNA,用于Allumila基因芯片分析,每组实验重复2次,并用荧光定量PCR验证表达有差异的基因。[结果]基因芯片结果表明,PDCD6敲除后共有133个基因表达有显著差异。其中,101个基因表达下调,32个基因表达上调。基因功能分析显示它们分别与细胞凋亡、增殖、周期、迁移和血管生成等有关。信号通路富集度显示MAPK信号通路激活,主要激活的基因有FGFR、MAP2K1/MEK1、MAP2K2/MEK2、MYC、FOS。RT-PCR验证结果显示,PDCD6敲除后CCND1、MYC、ANGPTL4、BMP2、CXCL16基因m RNA的表达比对照组明显下调,与基因芯片的结果相同。[结论]PDCD6可能通过上调CCND1、MYC、ANGPT4、BMP2和CXCL16基因,激活MAPK信号通路,从而促进卵巢癌细胞的增殖和转移。
[Purpose] PDCD6 is the abbreviation of programmed cell death 6. In recent studies it has been shown that PDCD6 can promote the proliferation and migration of ovarian cancer cells in a certain extent. However,its exact further molecular mechanism needs further research. [Methods] According to the previous experimental results,HO-8910PM cells treated with pGC- SIL-GFP-shRNA-PDCD6 virus solution （MOI2） for 72h as the experimental group,and HO- 8910PM cells treated with pGCSIL-GFP-shRNA-mock virus solution （MOI2） as the control group. Then,total RNA of HO-8910PM cells was extracted,amplified cRNA marked biotin,which allomixis with Illumina chip to scan, and analyzed by Illumina bead studio application software and DAVID database. Experiment was repeated twice. Several significant different expression genes were screened which related to tumor ceils proliferation, apoptosis and migration, and verified by RT-PCR. [Results] Gene chip suggested that there was a clear difference in 133 genes（diffseore less than -13 or greater than 13）,in detail,expression of 101 genes downregulated and expression of 32 genes upregulated. Gene function analysis revealed they correlated with apoptosis ,proliferation,cycle,migration,angiogenesis and so on. Signaling pathway enrichment analysis showed that the classic MAP kinase pathway （ERK1/2, extraeellular signal-regulated kinase） had been activated. The main actived genes included FGFR,MAP2K1/MEK1 ,MAP2K2/MEK2,MYC and FOS. RT-PCR results showed that the mRNA expressions of CCNDI, MYC, ANGPTL4, BMP2 and CXCL16 were significantly lower than those in control group,which the result was similar as the chip＇s. [Conclusion] PDCD6 might promote ovarian cancer cell proliferation and metastasis by upregulating the CCND1,MYC,ANGPT4,BMP2 and CXCL16 gene expression through the MAPK signaling pathway.
出处
《肿瘤学杂志》
CAS
2015年第2期113-122,共10页
Journal of Chinese Oncology
基金
国家自然科学基金项目(81101986)
浙江省自然科学基金项目(Y2111317)
