CXCR4抑制剂LPS对胆管癌细胞株QBC939生物学行为的影响

Inhibition of CXCR4 by LPS and the Biological Behavior Changes of Cholangiocarcinomal Cell QBC939

目的:探讨人胆管癌细胞株QBC939中CXCR4基因的表达,及LPS对CXCR4表达及生物学行为的影响。方法:RT-PCR和Western blot法检测胆管癌细胞株QBC939中CXCR4 mRNA及蛋白的表达,与不同浓度LPS干预后CXCR4 mRNA及蛋白表达的变化;MTT法和平板克隆形成试验检测LPS对QBC939细胞生长及克隆形成的抑制作用;趋化试验检测LPS对QBC939细胞趋化侵袭能力的影响。结果:QBC939细胞中有CXCR4 mRNA及CXCR4蛋白的表达明显,LPS能有效的抑制QBC939细胞中CXCR4 mRNA及CXCR4蛋白的表达,且具有浓度相关性。不同浓度的LPS对胆管癌细胞株QBC939的增殖影响不同,24h内各浓度LPS对细胞增殖均无明显抑制,LPS浓度为0.1μg/mL时,48h内无明显影响,为1.0μg/mL、5.0μg/mL时,QBC939细胞生长受到明显抑制,72h各浓度LPS对细胞增殖均有明显抑制作用;平板克隆形成试验显示对照组细胞培养第10天即可见明显克隆体形成,至第14天时克隆体数目多、体积大。而试验组细胞于培养第13天时才出现克隆体,且克隆体数目少、体积小。CXCL12(SDF-1)对OBC939细胞有明显的趋化作用,对照组细胞趋化率为53.5%,LPS各浓度组细胞趋化率明显低于对照组。结论:胆管癌细胞株OBC939中CXCR4 mRNA及CXCR4蛋白呈阳性表达;CXCR4抑制剂LPS能有效抑制胆管癌细胞株QBC939的生长与转移能力,CXCR4可能成为胆管癌基因治疗的靶点之一。

Objective： To explore the expression of CXCR4 gene in the cholangiocarcinoma cell line QBC939 and the biological behavior changes of the cells after treatment with lipopolysaccharide. Methods： RT-PCR and Western-blot methods were performed to detect the expression of CXCR4 mRNA and CXCR4 protein in QBC939 cells, before or after being treated with LPS, respectively. The inhibition effect of tumor cell growth and clone formation after LPS administration in QBC939 cells were detected by the MTT and flat clone formation experiment. The infestation test was used to detect infestation ability in QBC939 cells after treatment with LPS. Results： CXCR4 mRNA and protein were positively expressed in cholangiocarcinoma cell line QBC939. CXCR4 mRNA and protein in QBC939 cells expressed differently when exposed in different LPS concentration gradients. MTT results showed the different effects of cell growth after treatment with LPS. Cells were not affected at 24 or 48 hour time points when LPS was at or below 0.1 g/mL. However, when LPS concentration reached up to 1.0 g/mL or 5.0 g/mL, there were definite depressive effects in the cells. After 72 hours, even with LPS concentration as low as 0.1 g/mL, the depressive effect remained. In the flat clone formation test, tumor cell clones formed in 10 days in the control group, while the test groups took about 12 days to form. CXCL12 （SDF-1） can induce QBC939 cells to migrate. The migration rate in the control group was （53.723%±1.812）, while the migration rates of the test groups were lower （P〈0.05）. Conclusion： QBC939 cell growth and the ability to metastasize could be suppressed by LPS. CXCR4 gene could become a promising therapeutic device for cholangiocarcinoma.