摘要
目的研究不同激活状态的巨噬细胞培养上清液对7721肝癌细胞株(SMMC7721)增殖和迁移的影响。方法人类单核白血病细胞(THP-1)经佛波酯(PMA)诱导成为巨噬细胞(UaM),再分别采用脂多糖(LPS)诱导成为经典活化的巨噬细胞(M1),白细胞介素(IL-4+IL-13)诱导成为替代性活化的巨噬细胞(M2);用ELISA法检测其IL-10和IL-12的表达水平,定量PCR法检测TNF-α、CCL3、iNOS、AMAC-1、CCL22和Arg-1的表达;用CCK8法和Transwell试验检测UaM、M1和M2培养上清液对SMMC7721增殖和迁移的影响。结果 M1诱导组IL-12、TNF-α、CCL3和iNOS表达升高,M2诱导组IL-10、AMAC-1、CCL22和Arg-1表达升高。M1诱导组SMMC7721细胞增殖活力均明显低于UaM组和M2组(F=12.135,P<0.01);UaM和M2诱导组SMMC7721细胞的增殖活力在浓度比为1∶1时无统计学差异(F=5.356,P=0.293),在其他浓度组均存在统计学差异(P<0.01);SMMC7721在各组间穿过Transwell基底膜的细胞数量均存在差异(F=105.442,P<0.01)。结论 M1培养上清液能抑制SMMC7721的增殖和迁移,M2培养上清液能促进SMMC7721的增殖和迁移。
Objective To explore the influence of culture supernatant of macrophages in different activation modes on proliferation and migration of hepatoma cells line SMMC7721. Methods Human mononuclear leukemia cells THP-1 were induced by phorbol myristate acetate （PMA） to differentiate into maerophages （UaM） which was further induced into classically activated macrophage （ M1 ） by LPS and alternatively activated macrophage （M2） by interleukin （IL-4 and IL-13）. The phenotypic characteristics of differently activated macrophages were identified by ELISA for determination of IL-10 and IL-12 and quantitative RT-PCR for TNF-c~, CCL3, iNOS, AMAC-1, CCL22 and Arg-1. The effects of culture supernatant of macrophages in different activation modes on the proliferation and mi- gration of SMMC7721 cells were detected by cell counting Kit-8 （ CCK8 ） and Transwell invasion assay. Results 1L-12, TNF-a, CCL3 and iNOS were highly expressed in M1, while IL-IO, AMAC-1, CCL22 and Arg-1 were highly expressed in M2. The proliferation activity of SMMC7721 cells induced by M1 culture supernatant was lower than that of M2 and UaM culture supernatants （ F = 12. 135, P 〈0.01 ）. Under the different conditions of volume ratio of culture supernatant to RPMI1640 culture medium, i.e. , 1： 1, 2： 1,4： 1, 8： 1, the proliferation activities of SMMC7721 cells induced by UaM and M2 showed statistical difference （ P 〈 0.01 ） except for the volume ratio of 1 ： 1 in which no difference between UaM and M2 was found （ F = 5. 356, P = 0. 293 ）. The numbers of cell passing through the basement membrane in Transwell assay showed statistically significant difference among UaM, M1, M2 and control groups （ F = 105. 442, P 〈 0.01 ）. Conclusion The culture supernatants of M1 may inhibit the proliferation and migration of SMMC7721 cells. By contrast, the culture supernatants of M2 showed promoting effect on proliferation and migration of SMMC7721 cells.
出处
《临床检验杂志》
CAS
CSCD
北大核心
2013年第2期105-108,共4页
Chinese Journal of Clinical Laboratory Science
基金
国家自然科学基金(81260302)
