Antineoplastic effect of a group of cycloart-one triterpene compound
A technology of cyclic jackfruit alkane and composition, which is applied in the field of cyclic jackfruit alkane triterpenoids to achieve the effects of preventing damage and scavenging free radicals
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Embodiment 1
[0027] Cytotoxicity test:
[0028] 1. Materials: human hepatoma cell line (HepG2), human acute promyelocytic leukemia cell line (HL-60), all obtained from ATCC (American Type Culture Collection), and drug-resistant human hepatoma cell line (HepG2-R) is Doxorubicin-resistant cells (a gift from City University of Hong Kong). Mouse hepatocytes were isolated from the liver of Kunming mice (purchased from the Experimental Animal Center of Guangzhou University of Traditional Chinese Medicine, animal qualification certificate number: Yuejian Zhengzi No. 2001A054) and obtained through primary culture. Medium RPMI1640, DMEM, and fetal bovine serum were all purchased from Gibico. MTT (3-(4,5-DIMETHYLTHIAZOL-2-YL)-2,5-diphenyl tetrazolium bromide) is a product of AMRESCO, subpackaged by Shanghai Bioengineering Co., Ltd., batch number: 0481B50. Adriamycin (doxirubicine), product of Sigma Company.
[0029] 2. Primary culture of mouse hepatocytes: Disinfect and expose the liver after anest...
Embodiment 2
[0035] Cell morphology test (cell cycle and apoptosis):
[0036] 1. Method: In a 35×10mm culture dish containing HepG2 and HL-60 cells, administer the drug separately. After a period of time, add 400ul AO / EB staining solution (100ug / ml AO solution, 100ug / ml EB solution, use PBS Preparation), mix well, and immediately observe under a 40×10 fluorescent inverted microscope. AO solution can stain both living cells and dead cells, while EB solution only stains cells that have lost membrane integrity, and living cells still show a uniform green color. Early apoptotic cells appear green with bright green spots in their nuclei due to chromatin condensation and nuclear lysis. Late apoptotic cells can be stained orange by EB, but unlike dead cells, the nuclei of late apoptotic cells appear condensed and often lysed. Necrotic cells stain orange but have live-cell-like nuclear morphology and do not exhibit condensed chromatin.
[0037] 2. Results: The effects of the three compounds on ...
Embodiment 3
[0040] 1. Materials: RNase A (product of USB Company), 81.4 units / mg, batch number: 110266-007; PI (propidiumiodide) (product of Sigma Company, batch number: 042k3655).
[0041] 2. Methods: HepG2 and HL-60 cells were cultured by conventional methods, collected after administration, washed twice with PBS and fixed overnight with 70% alcohol. Centrifuge at 1000rpm / 10min to remove ethanol, wash twice with PBS containing 1% fetal bovine serum, resuspend with 2ml of PBS containing 1% fetal bovine serum, add RNase enzyme (final concentration 0.5mg / ml) to digest for 1 hour, and then Add PI (2.5ug / ml) for staining, place on ice for 20min, and then detect with flow cytometry.
[0042] 3. Results: The three compounds can stop HepG2 in the G2 / M phase at 30um in a time-dependent manner (Table 2). At 20um for 12 hours, HL-60 cells were arrested in G2 / M phase with obvious apoptosis (Table 3) (Figures 16-19).
[0043] compound
[0044] compoun...
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