SW620 cells were transfected with NF-Kappa B p65 siRNAs as the treatment group, negative control siRNA-A as negative control group or PBS Lumacaftor nmr as normal control group using Lipofectamine 2000 for 6 hours at 37°C in 6-well plates. The nuclear

NF-Kappa B p65 protein, the relative CXCR4 mRNA level, the total cell CXCR4 protein level and the cell surface level of CXCR4 was determined by ELISA, TaqMan RT-PCR reaction, western blot and flow cytometry. Results: The nuclear NF-kappa B p65 OD value of the normal control group, negative control group and the treatment group was 0.298 ± 0.044, 0.308 ± 0.034 and 0.085 ± 0.019. The relative CXCR4 mRNA level of the normal control group, negative control group and the treatment group was 1 ± 0.04, 0.96 ± 0.05 and 0.17 ± 0.01. The percentages of CXCR4 positive cells of the normal control group, negative control group and the treatment group was 97.6% ± 2.3%,

95.2% ± 2.8% and 8.9% ± 2.0%. The nuclear NF-kappa B p65 protein level, the relative CXCR4 mRNA level, the total cell CXCR4 protein level and the percentages of CXCR4 positive cells of the treatment group were much lower than those of the normal control and the negative control group. Conclusion: CXCR4 is regulated by NF-kappa B pathway in colorectal carcinoma cell line SW620. Acknowledgements: This study was supported by National Natural Science Foundation of China, No. 81272640; Guangdong Science and Technology Program, No. 2010B031200008 and No. 2012B031800043. Key Word(s): 1. old CXCR4; 2. NF-kappa

B; 3. colorectal carcinoma; Presenting Author: XIUQING WEI selleck compound Additional Authors: HUIXIN XIN, WEI MAO, YUNWEI GUO, BIN WU Corresponding Author: XIUQING WEI Affiliations: Department of Digestive Disease, Third Affiliatted Hospital of Zhongshan University Objective: CC chemokine receptor 7 (CCR7) and NF-kappa B pathway are both upregulated and play an important role in lymph node metastasis in human colon cancer. The aim of this research was to study whether CCR7 is regulated by NF-kappa B pathway. Methods: NF-kappa B p65 siRNAs and negative control siRNA-A were commercially designed by Santa Cruz biotechnology, inc. SW620 cells were transfected with NF-Kappa B p65 siRNAs as the treatment group, negative control siRNA-A as negative control group or PBS as normal control group using Lipofectamine 2000 for 6 hours at 37°C in 6-well plates. The nuclear NF-Kappa B p65 protein, the relative CCR7 mRNA level, the total cell CCR7 protein level and the cell surface level of CCR7 was determined by ELISA, TaqMan RT-PCR reaction, western blot and flow cytometry. Results: The nuclear NF-kappa B p65 OD value of the normal control group, negative control group and the treatment group was 0.298 ± 0.044, 0.308 ± 0.034 and 0.085 ± 0.019. The relative CCR7 mRNA level of the normal control group, negative control group and the treatment group was 1 ± 0.06, 0.99 ± 0.09 and 0.17 ± 0.02.