CXCR4在不同转移潜能肝癌细胞中的表达及意义

The Expression and Significance of CXCR4 in Human Liver Cancer Cell Lines with Different Metastatic Potentials

目的研究趋化因子受体CXCR4在高、低转移潜能肝癌细胞中的表达,并探讨其在肝癌细胞增殖和侵袭转移过程中的作用。方法利用3B肝癌细胞株对人工基底膜穿透能力的差异获得不同转移潜能肝癌细胞亚系,并通过Transwell小室侵袭实验比较两细胞亚系的侵袭能力从而对其进行验证,应用RT-PCR和Western blot检测CXCR4在两细胞亚系中的表达。CCK8法检测两细胞亚系生长曲线和倍增时间,流式细胞仪检测两细胞亚系细胞周期和凋亡率。结果成功筛选出高、低转移潜能肝癌细胞亚系,并通过Transwell小室侵袭实验证实高转移潜能肝癌细胞的侵袭能力高于低转移潜能肝癌细胞;RT-PCR和Western blot均显示高转移潜能肝癌细胞在基因水平和蛋白水平的CXCR4表达均显著高于低转移潜能肝癌细胞;且两细胞亚系体外生长速度、倍增时间、细胞周期和凋亡率差异均具有统计学意义。结论上述结果表明CXCR4在肝癌进展中发挥重要的作用,可能为未来肝癌的诊疗提供理论依据。

Objective To detect the expression of CXCR4 in human liver cancer cell lines with different metastatic potentials and to investigate the effect of CXCR4 on proliferation and metastasis of human liver cancer.Methods Matrigel invasion assays were applied to screen cell subclones of human liver cancer cell lines（3B） with different metastatic potentials,and transwell assays were applied to observe the cell motility and invasiveness for verifying the above result.RT-PCR and Western blot were used to detect the gene and protein expression of CXCR4 in cell subclones with different metastatic potentials.CCK-8 assay was applied to detect cell growth curve and doubling time;flow cytometry was used to test cell cycle and apoptosis rate.Results Two cell subclones with high metastatic potential and low metastatic potential were obtained through invasion assays,and it was verified that invasion ability in cell subclones with high metastatic potential is greater than that in cell subclones with low-metastatic potential by transwell test.The expression level of CXCR4 in mRNA and protein level in cell subclones with high metastatic potential were significantly higher than those in cell subclones with low metastatic potential.There were significant differences between both cell subclones in cell proliferation,doubling time,cell cycle and apoptosis rate.Conclusion Overexpression of CXCR4 is highly related to metastatic potential of human liver cancer cells.CXCR4 may play an important role in malignant progression of human liver cancer.It may be an important predictor and show us some new therapeutic approaches to treat liver cancer.