Heat shock protein 70 chaperoned alpha-fetoprotein in human hepatocellular carcinoma cell line BEL-7402

Abstract

Aim: To investigate the interaction between heat shock protein 70 (HSP70) and ?-fetoprotein (AFP) in human hepatocellular carcinoma (HCC) cell line BEL-7402.

Methods: The expression and localization of HSP70 and AFP in human HCC cell line BEL-7402 were determined by immunocytochemistry and indirect immunofluorescence cytochemical staining. The interaction between HSP70 and AFP in HCC cells was analyzed by immunoprecipitation and Western blot.

Results: Immunocytochemical staining detection showed that HCC cell BEL-7402 expressed a high level of HSP70 and AFP synchronously. Both were stained in cell plasma. AFP existed in the immunoprecipitate of anti-HSP70 mAb, while there was HSP70 in the immunoprecipitate of anti-AFP mAb.

Conclusion: HSP70 chaperones AFP in human HCC cell BEL-7402. The interaction between HSP70 and AFP in human HCC cell can be a new route to study the pathogenesis and immunotherapy of HCC.

Figure 1

Expressions of HSP70 and AFP in HCC cell line BEL-7402 by immunocytochemistry and immunofluorescence cytochemistry, ×400. A: HSP70 positive expression in cytoplasm; B: AFP immunostaining in cytoplasm; C: HSP70 immunostaining in cytoplasm; D: AFP green immunofluorescence in cytoplasm.

Figure 2

Analysis of immunoprecipitate of anti-HSP70 antibody and anti-AFP mAb by SDS-PAGE and Western blot (lane 1: precipitated with anti-HSP70 antibody; lane 2: precipitated with anti-AFP mAb; lane 3: precipitated with anti-HSA antibody; lane 4: cell lysates supernatants; M: molecular mass markers).