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目的研究核因子-κB(NF-κB)亚单位p65反义寡核苷酸(ASODN)对人胰腺癌细胞细胞间粘附分子-1(ICAM-1)表达及侵袭转移能力的影响。方法设计合成p65ASODN,体外转染人胰腺癌细胞株PANC-1,用流式细胞仪和荧光显微镜检测转染效率,用RT-PCR检测p65及ICAM-1mRNA的表达,用四甲基偶氮唑盐(MTT)法和细胞侵袭实验分析细胞增殖能力和侵袭转移能力的改变。结果p65ASODN可成功转染PANC-1细胞;转染后,p65及ICAM-1mRNA表达明显减弱(P<0.01);细胞增殖活性和侵袭转移能力明显下降(P<0.01)。结论脂质体介导的p65ASODN能有效抑制p65基因的表达,从而降低细胞的侵袭转移能力,p65ASODN可能主要通过下调ICAM-1基因的表达发挥抗侵袭转移的作用。
Objective To investigate the effect of p65 antisense oligonucleotide (NF-κB) subunit ASODN on the expression of ICAM-1 and its invasion and metastasis in human pancreatic cancer cells. Methods p65ASODN was synthesized and transfected into human pancreatic cancer cell line PANC-1 in vitro. The transfection efficiency was detected by flow cytometry and fluorescence microscopy. The expression of p65 and ICAM-1 mRNA was detected by RT-PCR. Salt (MTT) assay and cell invasion assay were used to analyze the changes of cell proliferation and invasion and metastasis. Results p65ASODN could be successfully transfected into PANC-1 cells. After transfection, the expression of p65 and ICAM-1 mRNA was significantly decreased (P <0.01), and the cell proliferation activity and invasion and metastasis were significantly decreased (P <0.01). Conclusion The p65 ASODN mediated by liposome can effectively inhibit the expression of p65 gene and thus reduce the invasion and metastasis of cells. P65 ASODN may play an important role in anti-invasion and metastasis by down-regulating the expression of ICAM-1 gene.