摘要目的 观察ARHI基因转染胰腺癌PANC1细胞后对趋化因子及其受体相关基因mRNA表达的影响.方法 采用脂质体法将表达ARHI基因的质粒pLNCX2-ARHI-EGFP、空质粒pLNCX2-EGFP转染胰腺癌PANC1细胞,应用G418筛选稳定转染细胞株.采用基因芯片RT2 ProfilerTMPCR Array行实时定量PCR,分析转染细胞的基因表达,包括84个趋化因子及受体相关基因,采用Real-time PCR法验证与血管生长相关基因mRNA的表达.结果 ARHI转染组细胞有36个基因mRNA表达下调,9个基因mRNA表达上调,39个基因mRNA表达变化无意义.其中肿瘤转移和侵袭相关基因CXCL12、CXCR4表达显著下调(<-6倍),MMP-2表达轻度下调(<-2倍);促肿瘤血管生成相关基因CCL2、CXCL1、CXCL2、CXCL8、CXCL12、CXCR4表达显著下调,CXCL3、CCR1、CCR2表达轻度下调;抗肿瘤血管生成相关基因CXCL9、CXCL10、CXCL11、CXCR3表达显著上调(>6倍);远端器官定位浸润和潜伏相关基因CXCL12、CXCR4、CCR7表达显著下调,CXCR5表达轻度下调;肿瘤免疫调节相关基因CXCL8、CXCR1、CCR7表达显著下调.Real-time PCR验证CXCL1、CXCL8、CXCR4、CXCR3结果与PCR Array检测结果一致.结论 ARHI基因可抑制胰腺癌细胞增殖、转移、血管生成、免疫调节相关趋化因子及其受体的基因表达.

abstractsObjective To investigate the effects of ARHI transfection on the chemokines and receptors related gene expression profile of PANC1 cells.Methods Plasmids expressing ARHI and empty plasmid were transfected into PANC1 cells, and the stably expressed cell lines were established by using G418.mRNA expression of chemokines and receptors related genes was detected by PCR Array.Real-time PCR was used to detect mRNA expression of the genes related vascular growth.Results In cells transfected with ARHI gene, the expression levels of mRNA of 36 genes were down-regulated, and 9 were up-regulated.Among the genes related to tumor metastasis and invasion CXCL12 and CXCR4 were significantly down-regulated (<-6 folds), and MMP-2 was slightly down-regulated (<-2 folds).Among the genes related to tumor angiogenesis, pro-angiogenesis genes including CCL2, CXCL1, CXCL2, CXCL8, CXCL12 and CXCR4 were significantly down-regulated, and pro-angiogenesis genes including CXCL3, CCR1 and CCR2 was slightly down-regulated.Anti-angiogenesis genes including CXCL9, CXCL10, CXCL11 and CXCR3 were significantly up-regulated (>6 folds).Among the genes related to the localization of distant organ infiltration and latency, CXCL12, CXCR4 and CCR7 were significantly down-regulated,and CXCR5 was slightly down-regulated.Among the gene with tumor immunity,CXCL8,CXCR1 and CCR7 were significantly down-regulated.Gene expression of CXCL1,CXCL8,CXCR4 and CXCR3 detected by Real-time PCR were consistent with PCR array.Conclusions ARHI gene inhibits the expression of chemokines and receptors related to tumor metastasis,angiogenesis and tumor immunity.