Experimental Study Of The Expressions Of HPA And ICAM-1 In The Colorectal Cancer Cells Treated By SDF-1 And CXCR4

Objective:To investigate the effects of SDF-1 and CXCR4 on the expressions of HPA and ICAM-1 in the colorectal cancer cells in vitro and vivo.To investigate some molecular mechanisms in colorectal liver metastasis treated by SDF-1 and CXCR4.To prove some valuable references for the transfer of tumor immunotherapy and anti-tumor metastasis therapeutic drug development.Methods:(1)The Lovo and SW480 human colorectal cancer cells in logarithmic growth period were made into single cell suspension ,and then co-cultured with different concentrations of SDF-1 for 12 and 24 or 48 hours. The expressions of HPA and ICAM-1 were detected by semiquantitative RT-PCR or immunocytochemical method respectively.(2)The single cell suspension was co-cultured with SDF-1 and AMD3100 for 24 or 48 hours. The expressions of HPA and ICAM-1 were detected by semiquantitative RT-PCR or immunocytochemical method respectively.(3) 18 nude mice model of colorectal cancer liver metastasis were established by splenectomy technique,Then mice were randomly divided into 3 groups:control group(saline solution) ,treatment group of low concentration AMD3100(0.1mmol/L,0.1ml/d) and treatment group of high concentration AMD3100(0.15mmol/L, 0.1ml/d ) . The expressions of HPA and ICAM-1 were examined by immunohistochemical technique.Result: (1) In Lovo and SW480 cells , After co-culturing with different concentrations of SDF-1(50 ng/ml,100 ng/ml,200 ng/ml )for 12 or 24 hours , the expressions of HPA and ICAM-1 mRNA were increased. Comparing with the same time, the expressions of HPA and ICAM-1 mRNA in the groups of 100 ng/ml and 200 ng/ml were higher than the group of 50 ng/ml and control group, and it was showed no significant difference between groups of 100ng/ml and 200ng/ml. Compared 12 to 24 hours with the same concentration of SDF-1, after co-culturing 24 hours with different concentrations of SDF-1, both the expressions of HPA and ICAM-1 mRNA were higher than 12, the control groups showed no significant difference.(2) In Lovo and SW480 cells , After co-culturing with SDF-1 and AMD3100 for 24 hours , the expressions of HPA and ICAM-1 mRNA in SDF-1(100 ng/ml)group were higher than other groups . There was significantly difference between SDF-1(100 ng/ml)group and other groups,and it was showed no significant difference between SDF-1(100 ng/ml) and AMD3100(1ug/ml)group and control group.(3) In Lovo and SW480 cells , After co-culturing with SDF-1 and AMD3100 for 48 hours , the expressions of HPA and ICAM-1 protein in SDF-1(100 ng/ml)group were higher than other groups . There was significantly difference between SDF-1(100 ng/ml)group and other groups,and it was showed no significant difference between SDF-1(100 ng/ml) and AMD3100(1ug/ml)group and control group.(4)In the liver metastases tumor tissues, the expressions of HPA and ICAM-1 protein(IOD values)were significantly difference between each groups of the control group,low-dose treatment group,high-dose treatment group.Conclusion:1. Both HPA and ICAM-1 were expressioned in colorectal cancer cell lines Lovo and SW480 and nude mice model of colorectal cancer liver metastasis . 2. The expressions of HPA and ICAM-1 were increased in the colorectal cancer cells that was treated by SDF-1, and it had some relations with the SDF-1 concentration and time .3. The expressions of HPA and ICAM-1 treated by SDF-1 were inhibited by AMD3100. AMD3100 is a CXCR4 specific inhibitor, it can block SDF-1/CXCR4 axis.4. Both HPA and ICAM-1 were reduced in the liver metastases tumor tissues treaded by AMD3100.it had the effection of inhibition the tumor metastasis by reduced tumor cells adhesion and invasion...