Does streptozotocin [STZ] exert an influence on rat hepatoma H35-19 cell line?
The authors investigated the effect of streptozotocin (STZ) in low--micromoles (up to 500 microM)--or higher --millimoles (1-10 mM)--concentrations in culture media of the H35-19 cell line. Up to 500 microM, STZ did not show any cytotoxic or cytostatic action in the investigated cells; on the contrary, it triggered an "improved growth" of these cells, as an antibiotic effect of the drug was observed. The concentration of 1-10 mmoles of STZ in the medium inhibited proliferation and viability of the studied cells. This action depended (proportionally) on drug concentration and time (up to 72 h) of experiment. Statistical analysis of the results obtained by four methods: staining with MTT, neutral red (NR) or crystal violet (CV) and Biirker chamber counting (BC), demonstrated no significant difference in STZ impact between 48 h and 72 h of incubation, according to the Benferoni post-hock test. The results obtained by MTT showed an extremely high statistical significance (p<0.001) of the effect of concentration on the results, with a non-significant interaction (p=0.2236) and general time effect (p=0.3600). An extremely significant (p<0.001) interaction of the effect of time and concentration was observed in the results obtained by neutral red method, whereas a significant effect of general time and concentration was also observed, but according to [17] it is difficult to explain. The results obtained by crystal violet staining showed a highly statistical significance (p<0.001) in time and concentration effect on the data, without a significant interaction between the above-mentioned factors (p>0.05). Cell counting in a Biirker chamber demonstrated a highly significant time and concentration effect on the results, but the interaction was mildly significant (0.01<p<0.05); the time and concentration effect are said to be difficult to interpret. The main STZ effect exerted on proliferation and growth inhibition was noted after 48 h and 10 mM of STZ. Morphological studies after 48 h and 5 mM STZ were performed, since these experimental conditions allowed for observing changes induced by the drug, when not all cells were destroyed by streptozotocin. At the studied streptozotocin concentration, the death of these cells occurred mainly by necrosis.
Dabroś
Wojciech
W
Department of Clinical and Experimental Pathomorphology, Faculty of Medicine, Jagiellonian University, Kraków. mpdabros@cyf-kr.edu.pl
Klein
Andrzej
A
Tatar
Barbara
B
Holko
Przemysław
P
Kordowiak
Anna M
AM
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D006528:Carcinoma, Hepatocellular / Q000473:pathology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D022781:Hepatocytes / Q000187:drug effects* / Q000648:ultrastructure*; D008113:Liver Neoplasms / Q000473:pathology; D009336:Necrosis / Q000473:pathology; D051381:Rats; D013311:Streptozocin / Q000494:pharmacology*
D000903:Antibiotics, Antineoplastic; D013311:Streptozocin