5638
Manar Ahmed Ahmed Elhinnawi
Synthesis of Novel Hybrid Steroids with Possible Cytotoxicity and Pro-apoptotic Effects on Human Hepatocellular Carcinoma In vitro
Synthesis of novel hybrid steroids with possible cytotoxicity and pro-apoptotic effects on human hepatocellular carcinoma in vitro.
A series of pregnenolone derivatives were synthesized and evaluated for anti-cancer activity against hepatocellular carcinoma cell line (HepG2). The synthesized steroidal derivatives 3, 4, 5, 6, 7, 8a and 8b were assessed for their cytotoxic activities using MTT (3-(4,5-Dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide) assay. Apoptotic activity was evaluated using dual acridine orange/ethidium bromide staining method and DNA fragmentation assay. The expression level of the pro-apoptotic genes Bak and Bax as well as anti-apoptotic genes Bcl-xL and Bcl-2 were analyzed using quantitative real time PCR. The results showed that compounds 4 and 6 exhibited cytotoxic activity (IC50s, 36.97 ± 2.18 and 18.46 ±0.64 μM, respectively), whereas compounds 5 and 7 displayed weak cytotoxic activity (IC50s, 93.87 ± 8.30 μM and 93.48 ± 4.14 μM, respectively). All synthesized pregnenolone derivatives induced apoptosis through DNA fragmentation. Compounds 4 and 6 increased early and late apoptotic cell percentages while compounds 3, 5, 7 and 8b increased either early or late apoptotic cell percentage. At the gene level, compounds 3, 6 and 8b up-regulated the expression level of Bak gene. On the other hand, compounds 4, 5, 7 and 8a down-regulated the Bcl-2 expression level, besides, compounds 5, 7 and 8a down-regulated the Bcl-xL expression level. Compounds 5, 7, 8a and 8b raised the Bak/Bcl-xL ratio besides compound 8a elevated the Bax/Bcl-xL ratio whereas compound 5 increasedBax/Bcl-2 and Bak/Bcl-2 ratios. The present work introduced novel pro-apoptoticpregnenolone derivatives that act against HepG2 cells through apoptotic morphological changes, DNA fragmentation and the increase of pro-apoptotic/anti-apoptotic ratios of Bcl-2 family genes.
2019
M.Sc
Cairo
Science