摘要
目的探讨CXC族趋化因子（CXCL）5在肝癌细胞中的表达及其对细胞增殖、迁移和侵袭的影响。方法利用实时定量PCR（RT—PCR）和酶联免疫吸附试验（ELISA）分别检测4种转移潜能依次递增的肝癌细胞（HepG2、SMMC7721、MHCC97L和MHCC97H）中CXCL5的mRNA和蛋白水平；选取低表达CXCL5的HepG2细胞为实验对象，接受外源性CXCL5刺激；实验分4组：CXCL5浓度分别为0、0．1、1．0、10．0nmol／L；利用细胞计数试剂盒（CCK-8）法检测细胞增殖情况；利用Transwell小室和基质胶观察CXCL5对细胞迁移和侵袭的影响。采用SPSS16．0统计软件进行数据分析，多组均数之间比较采用单因素方差分析（ANOVA），两两比较采用LSD检验。结果HepG2、SMMC7721、MHCC97L和MHCC97H中CXCL5的mRNA相对表达量分别为：0．002％±0．000％、0．005％±0．000％、1．030％±0．070％、0．980％±0．190％（F=33．88，P〈0．01）；蛋白分泌量分别为：14．3±0．4、25．7±1．4、82．8±3．2、98．94-1．7（F：447．08，P〈0．01）。CCK-8结果示细胞增殖随CXCL5浓度升高而增强，但差异无统计学意义（F〈1．00，P〉0．05）；0、0．1、1．0、10．0nmol／L组迁移细胞数分别为（29±3）、（56±16）、（113±7）、（130±15）个（F=51．94，P〈0．01）；侵袭细胞数分别为（17．3±1．8）、（33．0±3．2）、（65．7±4．4）、（94．3±3．5）个（F=104．13，P〈0．01）。结论转移潜能高的肝癌细胞能够高表达CXCL5；转移潜能低的肝癌细胞在受到外源性CXCL5刺激后，细胞增殖、迁移和侵袭能力均升高，提示CXCL5可能与肝癌的转移有关。
Objective To explore the expression of CXC chemokine 5 （CXCL5） in liver cancer cells and its effect on cell proliferation, migration and invasion. Methods Real-time （RT）-PCR and enzyme-linked immunosorbent assay （ELISA） were used to detect the mRNA and protein levels of CXCL5 in 4 liver cancer cell lines with different metastatic potentials （in ascending order： HepG2, SMMC7721, MHCC97L and MHCC97H）. HepG2 with a low expression of CXCL5 was treated with CXCL5. There were four groups ： 0 nmol/L CXCL5, 0. 1 nmol/L CXCL5, 1.0 nmol/L CXCL5 and 10. 0 nmol/L CXCL5. Cell proliferation was evaluated by cell counting kit-8 （CCK-8） assay. Transwell chambers and basement membrane matrix （Matrigel） were used to observe the cellular migration and invasion. Statistical analysis was performed with SPSS 16.0. Statistical comparison of the results was made by analysis of variance （ANOVA）. Results The relative mRNA expression levels of CXCL5 in HepG2, SMMC7721, MHCC97L and MHCC97H were 0.002% ±0.000%, 0.005% ± 0.000%, 1.030% ±0.070% and 0.980% ± 0. 190% （ F = 33.88, P 〈 0.01 ） while their protein levels 14. 3 ± 0. 4, 25.7 ± 1.4, 82. 8 ± 3.2 and 98. 9 ± 1.7 respectively （F =447. 08, P 〈 0. 01 ）. The CCK-8 results showed that cell proliferation increased with the treatment of CXCL5, but no significant difference existed （ F 〈 1.00, P 〉 0. 05 ） , cell numbers of migration of 0, 0.1, 1.0, 10.0 nmol/L CXCL5 groups were 29 ± 3, 56 ± 16, 113 ± 7 and 130 ± 15 （ F = 51.94, P 〈 0. 01 ）, while cell numbers of invasion 17. 3 ± 1.8, 33.0± 3.2, 65.7 ± 4. 4 and 94. 3 ± 3.5 respectively （ F = 104. 13, P 〈 0. 01 ）. Conclusions Liver cancer cells with high metastatic potential have a higher expression of CXCL5. And exogenous CXCL5 can increase the proliferation, migration and invasion of liver cancer cells with low metastatic potential. Thus CXCL5 may be associated with the metastasis of liver cancer.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2012年第38期2716-2719,共4页
National Medical Journal of China
关键词
癌
肝细胞
细胞运动
肿瘤侵润
趋化因子
CXC
Carcinoma, hepatocellular
Cell movement
Neoplasm invasiveness
Chemokines, CXC
