Cell experiment:

Cells are harvested during logarithmic growth phase and seeded in 96-well plates at a density of 1.5×104 cells/mL of final volume 190 μL/well. After incubation of 24 h, 10 μL EFL3 solution of full range concentrations is added to 96-well plates. After 68 h treatment, 10 μL MTT of 10 mM is added to each well for 4 h of maintaining at 37℃. Then, the supernatant is removed and the crystals are dissolved with 100 μL anhydrous DMSO each well. Subsequently, cell viability is measured by Model 550 Microplate reader at 540 nm and 655 nm as reference filter. Experiments are carried out at least thrice. The 50% inhibitory concentration (IC50) is defined as the anticancer agent concentration causing 50% reduction in cell viability and calculated from the cytotoxicity curves. Cell survival is calculated with the following formula: survival (%)=(mean experimental absorbance/mean control absorbance) × 100%.

5,15-Diacetyl-3-benzoyllathyrol is one of the lathyrane diterpenoids, that has anti-cancer activity.

5,15-Diacetyl-3-benzoyllathyrol shows anticancer activity in vitro against lung cancer A549 cells and the IC50 values are 34.04±3.99 μM. Furthermore, 5,15-Diacetyl-3-benzoyllathyrol can induce apoptosis in A549 cells via the mitochondrial pathway including loss of mitochondrial potential and release of cytochrome c[1].

References:
[1]. Zhang JY, et al. Structure identification of Euphorbia factor L3 and its induction of apoptosis through the mitochondrial pathway. Molecules. 2011 Apr 15;16(4):3222-31.