The effect of R547, a cyclin-dependent kinase inhibitor, on hepatocellular carcinoma cell death

Authors : Betül HACIOĞLU, Gökhan KUŞ, Hatice Mehtap KUTLU, Selda KABADERE

Pages : 24-33

View : 20 | Download : 6

Publication Date : 2020-12-01

Article Type : Research Paper

Abstract :Hepatocellular carcinoma insert ignore into journalissuearticles values(HCC); is the third main cause of cancer-related death. Cyclin-dependent kinases insert ignore into journalissuearticles values(CDKs); and their cyclin partners regulate the cell cycle. Since inhibition of CDKs gives some guiding ideas for cancer studies, we aimed to determine the possible effects of R547, a cyclin kinase 1-2-4 inhibitor, on proliferation and apoptotic mechanisms of Hep G2 cells insert ignore into journalissuearticles values(human); and H-4- II-E cells derived from rat HCC. We determined in vitro survival rates with MTT assay, apoptosis with flow cytometry, morphological changes with confocal microscopy, and ultrastructural changes by transmission electron microscopy. Cisplatin was used as a positive control. After 24 h of culture with 0.1, 1, 10, 50, and 100 ?M doses of R547, the corresponding percentages of live Hep G2 cells were 101%, 94%, 93%, 89%, and 79% insert ignore into journalissuearticles values(P < 0.001);, respectively. However, with the same R547 doses the live Hep G2 cell percentages were 92%, 101%, 53.6% insert ignore into journalissuearticles values(P <0 .01);, 47.4% insert ignore into journalissuearticles values(P < 0.001);, and 41% insert ignore into journalissuearticles values(P < 0.001);, respectively, after 48 h. After 24 h of incubation with the same doses of R547, the survival percentages of live rat cells were 90%, 80% insert ignore into journalissuearticles values(P < 0.01);, 63% insert ignore into journalissuearticles values(P < 0.001);, 47% insert ignore into journalissuearticles values(P < 0.001);, and 43% insert ignore into journalissuearticles values(P < 0.001);, respectively. The percentages of surviving H-4-II-E cells were 96%, 85% insert ignore into journalissuearticles values(P < 0.01);, 46% insert ignore into journalissuearticles values(P < 0.001);, 44% insert ignore into journalissuearticles values(P < 0.001);, and 45% insert ignore into journalissuearticles values(P < 0.01);, respectively, after 48 h. Since R547 did not significantly affect Hep G2 cell survival in 24 h, experiments of apoptosis were carried out with H-4-II-E cells. The early apoptotic rates of 38% and 45% insert ignore into journalissuearticles values(P < 0.05 for both); after applications of 10 and 25 μM R547 insert ignore into journalissuearticles values(control: 4.1%);, respectively, indicated that R547 has an apoptotic effect on H-4-II-E cells in 24 h. The apoptosis morphology at 24 h of treatment was clearly observed with microscopic examinations. According to our results, it is obvious that R547 has antiproliferative action when compared to cisplatin.
Keywords : R547, hepatocellular carcinoma, Hep G2, H 4 II E, apoptosis, flow cytometry, microscopy