Inhibition of proteinase 3 (PR3) by suramin and fetal calf serum (FCS): effect of PR3 and suramin on Chinese hamster ovary cells (CHO-cells).
BACKGROUND
Proteinase 3 (PR3) is a lysosomal protease that is stored in azurophilic granules neutrophilic granulocytes and monocytes. A number of inhibitors for this proteinase are reported. Comprehensive studies on the inhibitory effect of suramin and heat treated fetal calf serum (deltaFCS) on PR3 have not been reported. It has been reported that PR3 is able to destroy the cytoskeletal integral proteins, but we have not find any reports which showed the effect of this protease on Chinese hamster ovary cells (CHO-cells) in culture medium. Suramin has proven to be useful as an antitumor drug, but there was not any report on the effect of suramin on CHO-cells.


METHODS
The effects of various concentrations of deltaFCS (from 0.5% up to 10%) and suramin (from 0.8 microM up to 100 microM) on PR3 and different concentrations of suramin (from 0.8 microM up to 1000 microM) on CHO-cells were investigated. Data analysis were performed by, Kolmogorov-Smirnov test, ANOVA test and Tukey HSD post tests.


RESULTS
Results showed that deltaFCS and suramin have an inhibitory effect on PR3 and these effects increased with increasing the concentration significantly (p < 0.01). PR3 with the concentration of 2.2 Unit/ml has no effect on CHO-cells. Although suramin with the concentration of less than 125 microM cell growth retarded for only a few hours, but with the concentration of 125 to 250 microM inhibit the cell growth for a week, and after that cells gain normal growth gradually. Suramin with concentration of more than 500 microM inhibited the cell growth completely.


CONCLUSIONS
Although suramin reversibly inhibit the PR3 activity but in concentration of less than 250 microM it had no long-term effect on CHO-cells. Therefore it can be used in the investigation of proteases. There were unknown components in deltaFCS, which cause the inhibition of PR3 activity. This finding is very important in PR3 production in culture medium. However CHO-cells are resistant to PR3 and suramin in low concentration.
Karam
Gholamreza Asadi
GA
Department of Biochemistry, Faculty of Medicine, Rafsanjan Medical University, Post code: 7719617996, Rafsanjan, Iran. asadi_ka@yahoo.com
Rasaee
Mohammad Javad
MJ
Mahmoodi
Mehdi
M
Khaksari
Mohammad
M
D000818:Animals; D016466:CHO Cells; D002417:Cattle; D049109:Cell Proliferation / Q000187:drug effects; D006224:Cricetinae; D004305:Dose-Response Relationship, Drug; D053146:Myeloblastin; D012697:Serine Endopeptidases / Q000187:drug effects / Q000494:pharmacology*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology*; D044967:Serum / Q000502:physiology*; D013498:Suramin / Q000494:pharmacology*
D015842:Serine Proteinase Inhibitors; D013498:Suramin; D012697:Serine Endopeptidases; D053146:Myeloblastin