In vitro characterization of TMPRSS2 inhibition in IPEC-J2 cells.
The transmembrane serine protease, TMPRSS2 is an important target in the treatment of seasonal influenza infections and contributes to prostate carcinogenesis and metastasis. In this study, the effect of the synthetic TMPRSS2 inhibitor I-432 on jejunal IPEC-J2 cell monolayers cultured on membrane inserts was characterized. Using a fluorogenic substrate, it was found that the apical addition of I-432 could suppress trypsin-like activity in the supernatants of IPEC-J2 cells. The inhibition of TMPRSS2 did not affect physiologically produced hydrogen peroxide levels in the apical and in basolateral compartments. Loss of expression of the TMPRSS2 serine protease domain (28 kDa) was also observed when cells were pre-exposed to I-432. Partial decrease in immunofluorescent signal intensities derived from the altered distribution pattern of TMPRSS2 was detected after a 48 h long incubation of IPEC-J2 cells with the inhibitor indicating the efficacy of TMPRSS2 inhibition via I-432 administration in vitro.
a Faculty of Veterinary Science , Department of Pharmacology and Toxicology, Szent István University , Budapest , Hungary.
Erzsebet
E
Pászti-Gere
a Faculty of Veterinary Science , Department of Pharmacology and Toxicology, Szent István University , Budapest , Hungary.
Eszter
E
Czimmermann
b Faculty of Pharmacy , Semmelweis University , Budapest , Hungary.
Gabriella
G
Ujhelyi
c 1st Department of Pathology and Experimental Cancer Research, Semmelweis University , Budapest , Hungary , and.
Peter
P
Balla
d Faculty of Pharmacy , Institute of Pharmaceutical Chemistry, Philipps University Marburg , Marburg , Germany.
Alexander
A
Maiwald
d Faculty of Pharmacy , Institute of Pharmaceutical Chemistry, Philipps University Marburg , Marburg , Germany.
Torsten
T
Steinmetzer
D000818:Animals; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D015394:Molecular Structure; D012697:Serine Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D013552:Swine
D015842:Serine Proteinase Inhibitors; D012697:Serine Endopeptidases; C421305:TMPRSS2 protein, human
3-Amidinophenylalanine; IPEC-J2 cells; TMPRSS2; extracellular H2O2; fluorogenic AMC substrate; trypsin-like serine protease