Abstract: ABSTRACT：Objective To detect the expression level of RKIP in human lung carcinoma cell lines and investigate the relationships between RKIP and cell metastasis. Methods The expression level of RKIP in human lung carcinoma cell lines was analyzed by RT-PCR. Construct and express human RKIP fusion protein and immunize New Zealand rabbit with the purified fusion protein to prepare polyclonal antiserum.The antiserum titer was identified by ELISA. The specifcity of antiserum and expression level of RKIP in human lung carcinoma cell lines was analyzed by western blotting. Results The fusion protein was expressed successfully. The prepared polyclonal antibody reacted specifically with RKIP in human cells. Downregulation of RKIP was detected in three lung carcinoma cell lines with metastasis, and it is more significant in A549 with higher metastasis. Conclusion The RKIP antiserum with high titer and good specificity could satisfy the requirement for studying on RKIP. Downregulation of RKIP has correlation with human lung carcinoma cell metastasis.

Key words: RKIP, Polyclonal antibody, Lung carcinoma