Application of a gastric cancer cell line (MKN-28) for anti-adenovirus screening using the MTT method.
We established a sensitive and accurate method for screening of anti-adenovirus agents using the 3-(4,5-dimetylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. MKN-28 cells, which are well-differentiated stomach adenocarcinoma cells, were used for adenovirus (ADV) infection and examined for the anti-ADV activities of several established anti-herpes virus agents. ADV-11 is the causative agent of respiratory and urinary infections. It frequently causes hemorrhagic cystitis in immunocompromised hosts. One laboratory strain and 4 clinical isolates of ADV-11 were examined, and found susceptible (in order of decreasing activity) to 2-amino-7-[(1,3-dihydroxy-2-propoxy)methyl]purine (S-2242), (S)-9-(3-hydroxy-2-phosphonylmethoxypropyl)adenine[(S)-HPMPA ], and (S)-1-(3-hydroxy-2-phosphonylmethoxypropyl)cytosine[(S)-HPMPC++ +]. On the other hand, ganciclovir and iododeoxyuridine were only weakly effective and dextran sulfate was ineffective. Our findings indicate that the MTT assay using MKN-28 cells is applicable to anti-ADV screening. The anti-ADV activity of (S)-HPMPA and (S)-HPMPC was confirmed, and, furthermore, S-2242 emerged as a highly potent and selective inhibitor of ADV-11.
Kodama
E
E
Department of Microbiology, Fukushima Medical College, Japan.
Shigeta
S
S
Suzuki
T
T
De Clercq
E
E
D000225:Adenine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000260:Adenoviruses, Human / Q000187:drug effects* / Q000302:isolation & purification; D000998:Antiviral Agents / Q000494:pharmacology*; D000077404:Cidofovir; D003596:Cytosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D004353:Drug Evaluation, Preclinical; D005562:Formazans / Q000378:metabolism; D015774:Ganciclovir / Q000494:pharmacology*; D006801:Humans; D007065:Idoxuridine / Q000494:pharmacology*; D063065:Organophosphonates; D009943:Organophosphorus Compounds / Q000494:pharmacology*; D011687:Purines / Q000494:pharmacology*; D013274:Stomach Neoplasms / Q000473:pathology; D013778:Tetrazolium Salts / Q000737:chemistry / Q000378:metabolism; D013844:Thiazoles / Q000737:chemistry / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000998:Antiviral Agents; D005562:Formazans; D063065:Organophosphonates; D009943:Organophosphorus Compounds; D011687:Purines; C092709:S 2242; D013778:Tetrazolium Salts; D013844:Thiazoles; D003596:Cytosine; C050127:9-(S)-(3-hydroxy-2-(phosphonomethoxy)propyl)adenine; C022616:thiazolyl blue; D000225:Adenine; D000077404:Cidofovir; D007065:Idoxuridine; D015774:Ganciclovir