Li Yue-guo, Zhang Peng
Department of Laboratory, Tianjin Medical University Cancer Institute and Hospital, Tianjin, China.
Zhonghua Gan Zang Bing Za Zhi. 2012 May;20(5):372-6. doi: 10.3760/cma.j.issn.1007-3418.2012.05.014.
To explain biological function of protein CCL15 in HCC cell lines. 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 metastasis cell lines HCCML3 to observe the alteration of biological function of HCCML3. The overexpression of CCL15 in high metastasis 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 less than 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 less than 0.05). The expression of MMP-9 in HCCML3 was higher than HCCML3-siCCL15 through Western blot. Some biological properties (migration, invasion, MMP-9) of HCCML3 transfected with siRNA-CCL15 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.
为解释蛋白质CCL15在肝癌细胞系中的生物学功能。通过逆转录聚合酶链反应(RT-PCR)和蛋白质免疫印迹法(Western blot)验证了肝癌细胞系中CCL15的不同表达水平。成功构建了小干扰RNA(siRNA)-CCL15的表达重组质粒,并将其转染至高转移细胞系HCCML3中,以观察HCCML3生物学功能的变化。通过验证试验证实了CCL15在高转移肝癌细胞系中的过表达。用siRNA-CCL15转染后,细胞侵袭实验中侵袭细胞的平均数量分别为657.9(HCCML3)和148.4(HCCML3-siCCL15)(t = 19.34,P<0.05)。在划痕实验中,迁移距离分别为(0.35±0.02)mm(HCCML3)和(0.82±0.03)mm(HCCML3-siCCL15)(t = 15.67,P<0.05)。通过蛋白质免疫印迹法检测发现,HCCML3中基质金属蛋白酶-9(MMP-9)的表达高于HCCML3-siCCL15。转染siRNA-CCL15的HCCML3的一些生物学特性(迁移、侵袭、MMP-9)降低。结果表明,CCL15可能通过基质金属蛋白酶调节和增强侵袭潜能这两条途径在肝癌细胞侵袭和转移中发挥重要作用。