The Study Aims To Investigate The Production And Mechanism Of CCL5 By Macrophages In U14 Cervical Cancer Cells Mice During

Objective:Our study aims to investigate the production and mechanism of RANTES(CCL5) by macrophages in U14 cervical cancer cells mice during infectionMethods:The U14 cervical cancer cells were injected in C57BL/b mice to induce tumor-bearing condition. Lipopolysaccharide(LPS) were injected in C57BL/b mice to induce infection.The protein expression of Regulated upon Activation, Normal T-cell Expressed, and Secreted/chemokine(C–C motif)ligand 5,(RANTES/CCL5) in the serum and the CCL5 m RNA expression in inflammatory cells were measured by Elisa and Q-PCR in four groups.Macrophages were induced in the tumor conditioned medium(TCM) which extracted from mice serum. The protein expression levels of CCL5,Prostaglandin E2(PGE2)and Cyclic adenosine monophosphate(c AMP) in the medium and the CCL5, PGE2 and c AMP m RNA expression in the macrophages were detected in different groups. In order to determine whether the inhibition was related to PGE2, selective Cyclooxygenase2( COX-2)inhibitor NS-398 was used to reverse this phenomenon and Protein kinase A( PKA) inhibitor H89 demonstrated the mechanism through blocking CAMP/PKA signaling pathway.Results:1 CCL5 protein and m RNA level in tumor-bearing mice was 151.34±35.355 pg/ml and 1,which is lower than the tumor-free mice 690.81±84.852pg/ml and 4.53±0.848, the difference was significant(P<0.05). PGE2 protein and m RNA level in tumor-bearing mice was 1198.49±82.731 pg/ml and5.75±0.777,which is higher than the tumor-free mice 186.84±25.455 pg/ml and 1, the difference was significant(P<0.05).CCL5 protein and m RNA level in tumor-free+LPS mice was 4049.34±141.421 pg/ml and 31.49±1.980,which is higher than the tumorbearing+LPS mice 1950.68±70.71 pg/ml and 12.11±2.828, the difference was significant(P<0.05). PGE2 protein and m RNA level in tumor-free+LPS mice was 676.5±70.003 pg/ml and 3.35±0.353,which is lower than the tumor-bearing+LPS mice 2550±381.837 pg/ml and 11.55±0.9491, the difference was significant(P<0.05).2 Macrophages were cultured in vitro using TCM derived from mice.CCL5 protein and m RNA level in tumor-bearing mice TCM was1625.70±176.776 pg/ml and 28.65±1.202,which is higher than the tumor-free mice TCM 27.43±2.828 pg/ml and 1, the difference was significant(P<0.05).PGE2 protein and m RNA level in tumor-bearing mice TCM was790.49±155.563 pg/ml and 1.67±0.247,which is higher than the tumor-free mice TCM 448.30±114.551 pg/ml and 1, the difference was significant(P<0.05). c AMP protein and m RNA level in tumor-bearing mice TCM was164.48±29.698 pg/ml and 1.59±0.268,which is higher than the tumor-free mice TCM 117.54±24.748 pg/ml and 1, the difference was significant( P <0.05).CCL5 protein and m RNA level in tumor-free+LPS mice TCM was10475.12±742.46 pg/ml and 212.01±5.734,which is higher than the tumor-bearing+LPS mice TCM 6375.09±530.33 pg/ml and 142.28±2.545, the difference was significant( P < 0.05). PGE2 protein and m RNA level in tumor-free+LPS mice TCM was 2437.54±95.459 pg/ml and 4.31±0.707,which is lower than the tumor-bearing + LPS mice TCM 3440.56±162.634 pg/ml and5.94±0.283, the difference was significant( P < 0.05). c AMP protein and m RNA level in tumor-free+LPS mice TCM was 339.53±13.435 pg/ml and4.055±0.353,which is lower than the tumor-bearing+LPS mice TCM541.50±41.719 pg/ml and 5.35±0.495, the difference was significant( P <0.05). 3.Using COX-2 inhibitor NS398 in the tumor-bearing+LPS mice, CCL5 protein and m RNA level was higher in 7690.58±268.70 pg/ml and158.95±8.910,PGE2 protein and m RNA level was lower in 2819.96±152.027pg/ml and 4.94±0.283),while c AMP protein and m RNA level was lower in464.57±7.778 pg/ml and 4.26±0.424. Using PKA inhibitor H89 in the tumor-bearing+LPS+NS398 mice, CCL5 protein and m RNA level was higher in 8375.09±520.33 pg/ml and 177.05±8.838,PGE2 protein and m RNA level was lower in 2650.34±35.355 pg/ml and 4.66±0.424), while c AMP protein and m RNA level was lower in 367.53±13.435 pg/ml and 3.11±0.707.Conclusions:TCM of U14 cells activated macrophages to release PGE2 to inhibit expression of CCL5 levels by CAMP /PKA signaling pathway.