What is More Function About Aloe Emodin?

Neuroblastoma (SJ-N-KP), colon adenocarcinoma (LoVo 109), and cervix epithelioid carcinoma (HeLa) cell lines (1 × 106) were cultured for different time periods in the presence of aloe emodin or drug-free medium. Cells were harvested, washed twice with PBS, and fixed with cold 70% ethanol at 4°C. After centrifugation of the samples, propidium iodide (50 μg/ml in PBS) and RNase were added to the pellet for 20 min at 37°C to determinate the effect of aloe emodin on the cell cycle dynamics. DNA fluorescence was measured by flow cytometry (EPICS XL; Coulter, Miami, FL) analysis according to a published method. To determinate drug uptake, SJ-N-KP, LoVo 109, and HeLa cells were cultured in the presence of 25μ M of Aloe emodin or in drug-free medium at 37°C or at 4°C or in presence of NaN3, for 24 h and then analyzed by flow cytometry.

Two-Photon Excitation Microscopy.
Neuroblastoma (IMR5), colon adenocarcinoma (LoVo 109), and cervix epithelioid carcinoma (HeLa) cell lines were seeded on microscope coverslips in 12-well plates and cultured with drug-free medium 24 h before treatment. Then aloe emodin was added at different concentrations. At different time points, cells were washed twice with PBS and examined by means of “fluorescence two-photon confocal microscopy.” Optical sections were acquired with a TPE architecture described in detail elsewhere.

Transmission Electron Microscopy Analysis.

Cells were cultured with different concentrations of aloe emodin or with drug-free medium. At 24 and 48 h cells were scraped, washed twice in PBS, and fixed overnight at 4°C in 3% glutaraldehyde in 0.1 M sodium cacodylate buffer (pH 6.9) and then processed according to Ciman et al. . Ultrathin sections, cut with an ultramicrotome (Ultracut; Reichert-jung), were observed with the transmission electron microscope (TEM 300; Hitachi) operating at 75 kV.

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