Martha Ervina (2021) Aktivitas Antikanker Dan Mekanisme Apoptosis Senyawa Steroid Dari Daun Mindi (Melia Azedarach L.) B Terhadap Sel T47d. Disertasi thesis, UNIVERSITAS AIRLANGGA.

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Abstract

The prevalence of cancer continues to increase and becomes the second leading cause of death, both globally and nationally. Especially for breast cancer, which has increase in numbers and shifted to be the third largest. The unavoidable side effects and the emergence of resistance of chemotherapies, make important to find high selective chemotherapy agents. Among anticancer mechanisms, the apoptosis is restoring the homeostatic ability of the cell pathways. Mindi (Melia azedarach L.), has grown in more than a third of the world, but has not used widely for traditional medicine. Previous research not only yielded the cytotoxic potential of various parts of M. azedarach and other Melia species against several cell lines, but also isolated steroids compounds from it. Meanwhile, therapy with estrogen and tamoxifen (competitive estrogen) are used in several stages of breast cancer treatments. Therefore, this study aims to examine the activities and anticancer mechanisms of steroids compounds from M. azedarach leaves against T47D cells. The activities were determined by IC50 cytotoxic, while the apoptosis and cell cycle inhibition were examined by flowcytometer and immunocytochemicals. The separation and isolation were carried out using various chromatography methods with cytotoxic guided bioassay. The isolation procedure and LC-MS analysis applied to the standardized plant obtained tannins, flavonoids, alkaloids, saponins, glycosides, and steroids. The extract was prepared by using 96% ethanol, while fractionation by liquid-liquid extraction produced n-hexane (MH), ethyl acetate (ME) and water (MW) fractions. Cytotoxic bioassay showed ME fraction as the most active with IC50 166.48 μg /mL, while doxorubicin 3.08 μg/mL. Furthermore, ME11 and ME12 subfractions showed more potent activity than the others (IC50 92.16 and 113.15 μg/mL). The HPLC optimization of the ME11 and ME12subfractions resulted in ME114A, ME127A and ME127C components (IC50> 500, 173.62,60.86 μg/mL, respectively). ME114A compound has molecular weight of 626.1373 with the formula C27H30O17; which is determined as quercetin-3-O-β-D-galactopyranosyl-(1-6)-β-Dgalactopyranoside. ME127A and ME127C were obtained as isomers steroids with molecular weight of 330.208 and the formula C21H30O3. This study concluded that two new bioactive derivates of pregnene-16-one steroids isomer, namely ME127A and ME127C, can be isolated from Mindi (M. azedarach L.) leaves the ethyl acetate fraction. They are 17-ethylene-3,4-dihydroxy-14-methyl-18-norandrostene- 16-one and 17-ethylene-3,4-dihydroxy-5-pregnene-16-one. They showed moderate cytotoxic activity, while apoptotic by increasing ratio BAX/bcl-2 (intrinsic pathway) and cell cycle inhibitions on subG1, G1 and G2 phases mechanisms against T47D cell. However, further researches are required to highlight its safety and usage in vivo

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