Transfer of anti-TFAR19 monoclonal antibody into HeLa cells by in situ electroporation can inhibit the apoptosis.
Electroporation has been successfully used for the introduction of DNA, RNA, oligonucleotide and protein into eukaryotic and prokaryotic cells for the transformation and expression of various gene products. TFAR19 (TF-1 apoptosis-related gene 19), also designated PDCD5 (Programmed Cell Death 5), is cloned as an increased expression gene during the apoptotic process of TF-1 cell induced by cytokine withdrawal. It facilitates rather than induces apoptosis in different cell lines. To explore its molecular mechanism, we successfully transferred the anti-TFAR19 monoclonal antibody into HeLa cells by in situ electroporation and observed the apoptosis process of HeLa cells induced by etoposide with flow cytometry. We demonstrate that the introduction of anti-TFAR19 antibody can suppress the apoptosis accelerating effect of TFAR19 in its natural environment. This study shows that TFAR19 may be a critical factor for apoptosis; and transfer of monoclonal antibody into mammalian cells by in situ electroporation is a useful method to study the function of endogenous factors.
Rui
Min
M
Laboratory of Medical Immunology, School of Basic Medical Science, Peking University, 38 Xueyuan Road, Beijing 100083, People's Republic of China.
Chen
Yingyu
Y
Zhang
Yingmei
Y
Ma
Dalong
D
D019138:Antibodies, Blocking / Q000494:pharmacology*; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins; D015153:Blotting, Western; D018274:Electroporation; D005260:Female; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D006367:HeLa Cells; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D008856:Microscopy, Fluorescence; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors* / Q000276:immunology
D019138:Antibodies, Blocking; D000911:Antibodies, Monoclonal; D051017:Apoptosis Regulatory Proteins; D005456:Fluorescent Dyes; D007074:Immunoglobulin G; D009363:Neoplasm Proteins; C117283:PDCD5 protein, human; D016650:Fluorescein-5-isothiocyanate