ABSTRACT
Objective To evaluate effects of tanshinone ⅡAon the invasion and migration of melanoma A375 cells,as well as on the mRNA and protein expression of CXC chemokine receptor type 7 (CXCR7).Methods In vitro cultured A375 cells were divided into 4 groups to be treated with tanshinone ⅡA at different concentrations of 1,2 and 4 mg/L,and 0.1％ dimethyl sulfoxide (DMSO) (control group) for 48 hours,respectively.Wound scratch assay and Transwell invasion assay were conducted to estimate the migratory and invasive abilities of A375 cells,respectively,and real-time fluorescence-based quantitative PCR (qRT-PCR) and Western blot analysis to determine the mRNA and protein expression of CXCR7 in A375 cells,respectively.Results After 48-hour treatment,the 1-,2-and 4-mg/L tanshinone ⅡA groups showed significantly decreased number of A375 cells crossing the polycarbonate membrane per high-power field (× 200) (71.00 ± 4.00,51.00-± 2.00 and 37.00 ± 3.61,respectively) in Transwell invasion assay,as well as decreased number of A375 cells migrating to the scratch zone (301 ± 3.00,253.00 ± 3.61 and 126.00 ± 7.00,respectively) in wound scratch assay,compared with the control group (105.33 ± 6.51,332.00 ± 6.24,respectively,all P ＜ 0.05).Additionally,qRT-PCR and Western blot analysis revealed that the mRNA and protein expression of CXCR7 was significantly lower in the 1-,2-and 4-mg/L tanshinone ⅡA groups than in the control groups (CXCR7 mRNA:0.63-± 0.04,0.44 ± 0.02 and 0.31 ± 0.01 vs.1.00 ± 0.02;CXCR7 protein:0.573 ± 0.015,0.416 ± 0.011 and 0.260-± 0.055 vs.0.9000 ± 0.010;all P ＜ 0.05).Moreover,inhibitory effects of tanshinone ⅡA on the migration and invasion of A375 cells,as well as on the mRNA and protein expression of CXCR7,were significantly enhanced with the increase of tanshinone ⅡA concentrations (all P ＜ 0.05).Conclusion Tanshinone ⅡA can inhibit the migratory and invasive abilities of melanoma A375 cells by down-regulating CXCR7 expression.
ABSTRACT
Secondary lymphoid tissue chemokine (CCL21) is a double-edged sword, which exerts antitumor, anti-infection immune response by binding to the receptor CCR7 on the surface of the multiple immune cells. However, a variety of tumor cells also express the receptor CCR7, the combination of CCL21 with CCR7promotes the invasion and metastasis of tumor cells, leading to the facilitation of tumor development. Therefore,exploring the mechanism(s) of tumor invasion and metastasis might be helpful for use of CCL21 as tumor gene therapy through blocking of CCL21's promotion of tumor invasion and metastasis.
ABSTRACT
Objective To explore the influence of lung IL-10 on interstitial dendritic cells in multiple organ dysfunction syndrome(MODS),and its role in immunological dysfunction.Methods The model of MODS was replicated by injecting zymosan into the peritoneal cavity of C57BL/6 mice.The mice were then randomly divided into normal groups as well as the groups of 3-6 hours,12-48 hours,5-7 days and 10-12 days post trauma.The level of CCR7 mRNA in lung was detected by RT-PCR and the proteins of CCR7 and IL-10 were immunohistochemically stained.The number of mononuclear cells in MHC-II positive peripheral blood was determined by flow cytometry.Results At the early stage of injury(3-6 hours),the number of IL-10 positive cells and the expression of CCR7 increased,but the number of mononuclear cells in MHC-II positive peripheral blood decreased.At the stage of 12-48 hours,the number of IL-10 positive cells and the expression of CCR7 continued to increase obviously,and that of mononuclear cells in MHC-II positive peripheral blood went on decreasing.At the stage of 10-12 days,the number of IL-10 positive cells increased dramatically compared with that in normal group,but the expression of CCR7 declined obviously in contrast to that in 12-48 hours,and the number of mononuclear cells in MHC-II positive peripheral blood decreased to the lowest level.Conclusions The expression of IL-10 in lung could inhibit the excessive immunological damage induced by dendritic cells on the one hand,and it probably leads to immunosuppression by reducing the CCR7 expression of dendritic cells and the number of mononuclear cells in MHC-II positive peripheral blood on the other hand.