Downregulation of Raf kinase inhibitory protein in human lung cancer cell lines / 基础医学与临床

ABSTRACT

Objective To detect the expression level of RKIP in human lung cancer cell lines and to investigate the relationships between RKIP and cell metastasis. Methods The expression level of RKIP in human lung cancer cell lines was analyzed by RT-PCR. To construct and express human RKIP fusion protein and to immunize New Zealand rabbit with purified fusion protein to prepare polyclonal antiserum.The antiserum was titered 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 successfully expressed. The prepared polyclonal antibody reacted specifically with RKIP in human cells. Downregulation of RKIP was detected in three lung cancer 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 meet the requirement for studying on RKIP. Downregulation of RKIP has correlation with human lung carcinoma cell metastasis.