CCR4对肝癌细胞索拉非尼放射敏感性及裸鼠成瘤的影响

Effect of CCR4 on sorafenib radiosensitivity and tumorigenesis of hepatocellular carcinoma cells in nude mice

目的探讨CC趋化因子受体4(CCR4)对肝癌细胞索拉非尼放射敏感性及裸鼠成瘤的影响。方法采用Westernblot检测CCR4在肝癌细胞系中的表达。利用慢病毒感染构建过表达及敲减CCR4的PLC/PRF/5和SMMC-7721稳转株,并采用Western blot进行验证。采用平板克隆形成实验检测CCR4对肝癌细胞放射敏感性的影响。通过裸鼠成瘤实验检测CCR4对肝癌细胞体内成瘤的影响。结果高转移肝癌细胞中CCR4呈高表达,低转移肝癌细胞中CCR4呈低表达。成功构建稳定过表达CCR4的PLC/PRF/5和敲减CCR4的SMMC-7721肝癌稳转株,过表达CCR4可逆转索拉非尼放疗对PLC/PEF/5的抑制作用,而敲减CCR4可增加SMMC-7721对索拉非尼放射敏感性。过表达CCR4可促进PLC/PEF/5裸鼠体内成瘤能力,而敲减CCK4可抑制SMMC-7721裸鼠体内成瘤能力,结论CCR4高表达能显著增强肝癌PLC/PEF/5细胞对索拉非尼放射抵抗及裸鼠体内成瘤能力,而敲减CCK4则可显著增加肝癌SMMC-7721细胞对索拉非尼放射敏感性并抑制其裸鼠体内成瘤能力。

Objective Fo investigate the effect of CC chemokine receptor 4(CCR4)on sorafenib radiosensitivity and tumorigenesis of hepatocellular carcinoma cells in nude mouse models.Methods Western blot was used to detect the expression of CCR4 in hepatocellular carcinoma cell line.Lentivirus was utilized to construct PLC/PRF/5 and SMMC-7721 cell lines stably overexpressing and silencing CCR4,which were verified by Western blot.The influence of CCH4 on the radiosensitivity of hepatocellular carcinoma cells was assessed by plate clone formation assay.The effect of CCR4 on the tumorigenesis in hepatocellular carcinoma cells in vivo was evaluated by tumorigenesis assay in nude mice.Results CCR4 was highly expressed in highly-metastatic hepatocellular carcinoma cells and lowly expressed in hepatocellular carcinoma cells with low metastases.The PLC/PRF/5 and SMMC-7721 cells,which stably overexpressed and silenced CCR4,were successfully established.Overexpression of CCR4 reversed the inhibitory effect of sorafenib radiotherapy on PLC/PEF/5,whereas knockdown of CCR4 could increase the radiosensitivity of SMMC-7721 to sorafenib.Overexpressing CCR4 could promote the tumorigenicity of PLC/PEF/5,whereas knockdown of CCR4 could inhibit the tumorigenicity of SMMC-7721 in nude mice.Conclusion CCR4 overexpression significantly reduces the radiosensitivity of PLC/PRF/5 and increases the tumorigenicity in nude mice,whereas knockdown of CCR4 considerably increases the chemosensitivity and radiosensitivity of SMMC-7721 and suppresses the tumorigenicity in nude mice.