Application of quinolizidine in preparing tumor treatment drugs
A technology of quinolizidine and tumor drugs is applied in the application field of quinolizidine alkaloids in the treatment of tumor drugs, and can solve the problems that the use has not yet been reported.
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Embodiment 1
[0021] Example 1, using the conventional MTT method to carry out tumor suppression experiments, the methods and results are as follows:
[0022] MTT colorimetry is a method to detect cell survival and growth. This method has been widely used in large-scale antitumor drug screening, and it is characterized by high sensitivity and economy.
[0023] 0.5-2×105 of each of the above tumor cells was placed in a 96-well plate, and the preparation prepared by aloperine, spartine and cytisine was diluted to an experimental concentration, added to each tumor cell and treated for 48 hours. Three replicate wells were set up for each experimental group, the negative blank control wells were no drug added, and the positive drug control wells were sophoridine. The 5 mg / ml MTT solution stored at -20°C was dissolved at room temperature in the dark, and 10 μl of MTT solution was added to each well of a 96-well plate to mix well, and placed in a cell culture incubator for 4 hours. Add 100 μl of...
Embodiment 2
[0028] Example 2, inducing tumor cell autophagy:
[0029] It was shown in Example 1 that aloperine, spartine and cytisine can inhibit the proliferation of tumor cells. This example further verifies the anti-tumor activity of aloperine, spartine and cytisine.
[0030] After the drug preparations of each experimental group in Example 1 acted on the tumor cells for 18 hours, 0.5 ml of cells were taken and added to a 1.5 ml EP tube, and 5 μl of acridine orange was added, at 37° C. for 15 min. Add 0.5ml of PBS, centrifuge at 1500rpm at 4°C for 5min, discard the supernatant, collect the cells, and wash twice with 1ml of PBS. The cells were resuspended in 100 μl PBS, and the cells were observed under a fluorescent inverted microscope. It was found that the nuclei of the drug experimental group were red and autophagy occurred. This shows that the pharmaceutical preparation of the present invention can inhibit the proliferation of tumor cells by inducing the autophagy of the above-men...
Embodiment 3
[0031] Example 3, inducing tumor cell apoptosis:
[0032] PARP shearing and ladder DNA agarose gel electrophoresis, they are important indicators and characteristics of apoptosis. After the drug preparations of each experimental group continued to act on the above tumor cells for 48 hours in Example 2, PARP shearing and ladder DNA agarose gel electrophoresis were detected by conventional Western blotting, and it was found that the drug preparations continued to act on the above tumor cells for 48 hours Afterwards, PARP shearing and ladder-shaped DNA appeared, and the results showed that the pharmaceutical preparation of the present invention can finally induce the above-mentioned tumor cell apoptosis and achieve the effect of treating tumor cells.
[0033] Within the soluble concentration range of aloperine, spartine and cytisine, the IC50 of the action concentration is greater than 5-10 times more than tumor cells, indicating that aloperine, spartine and cytisine have less t...
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