摘要
目的解析CeL15的生物学功能，验证其是否在肝癌侵袭、转移和复发中发挥作用。方法用RT-PCR、Westemblot分别检测CCL15在不同转移潜能人肝癌细胞株中的基因和蛋白表达水平并进行比较。用CCL15特异性的小干扰RNA质粒转染高转移潜能人肝癌细胞株HCCML3，得到CeL15低表达的细胞株HccML3-SiCCL15，通过划痕实验和Matrigel体外侵袭实验观察CeL15表达降低后对HCCML3细胞生物学行为的影响。数据用均数±标准差伍±s）表示，用SPSS11．0统计软件包进行t检验。结果CeL15在有转移潜能人肝癌细胞株HCCML3和MHCC97-L中高表达。划痕实验结果显示，HCCML3组和HCCML3siCCLl5组划痕两侧边缘之间的距离分别是（0．35士0．02）mm和（0．82±0．03）mm（t=15．67，P〈0．05）。Matrigel侵袭实验结果显示，HCCML3组和HCCML3-siCCLl5组平均每视野侵袭细胞分别为（657．9±22．9）个和（148．4±10．2）个（t=19．34，P〈0．05）。HccML3-SiccL15组细胞内基质金属蛋白酶（MMP-9表达水平明显低于HCCML3组。结论降低CCLl5表达后HCCML3细胞迁移和侵袭能力均明显降低，细胞内MMP-9表达水平降低。提示CCL15通过增强肝癌细胞的运动侵袭能力和上调MMP-9的分泌参与肝癌细胞的侵袭、转移过程。
Objective To explain biological function of protein CCL15 in HCC cell lines. Methods The different expression level of CCL15 among HCC cell lines was validated by RT-PCR and Western blot. The expression recombinant plasmid of siRNA-CCL15 was constructed successfully and transfected into high metastasic cell lines HCCML3 to observe the alteration of biological function of HCCML3. Results The overexpression of CCL15 in high metastasic HCC cell lines was confirmed by validation tests. After transfected with siRNA-CCL15, the average amounts of invaded cells in cell invasion assay were 657.9 （HCCML3） and 148.4（HCCML3-siCCL15） （t= 19.34, P〈0.05）. And in the scratch assay, the migrating distance were （0.35 ± 0.02） mm （HCCML3） and （0.82 ± 0.03）mm （HCCML3-siCCL15） （t= 15.67, P〈 0.05）. The expression of MMP-9 in HCCML3 was higher than HCCML3-siCCL15 through Western blot. Conclusion Some biological properties （migration, invasion, MMP-9） of HCCML3 transfected with siRNACCL15 were decreased. The results suggest CCL15 might play an important role in HCC cell invasion and metastasis through two paths of MMPs regulation and invasion potential strengthening.
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2012年第5期372-376,共5页
Chinese Journal of Hepatology
基金
国家自然科学基金（81101754）
国家高技术研究发展计划（8635计划,2011AA02A111）
关键词
癌
肝细胞
肿瘤转移
肿瘤侵润
CCL15
Carcinoma, hepatocellular
Neoplasm metastasis
Neoplasm invasiveness
CCL15
