Application of shikonin in preparing drug for treating lung cancer
A shikonin and drug technology, applied in the field of anti-tumor drug preparations, can solve the problems of cancer cell aging and anti-tumor effects, and achieve the effect of inducing cancer cell aging
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experiment example 1
[0022] Experimental example 1: Inhibitory effect of shikonin on the viability of lung cancer cells
[0023] When the cell growth state is good, inoculate 2000 cells / well in a 96-well plate and culture overnight. When the cell adhesion state is good, discard the original medium, and then follow the designed drug concentration: shikonin concentration 0 , 1, 2, 4, 6, 8μM for medium change administration, act on A549 cells for 72h, stop the culture, suck out the medium, add 100μl of 0.5mg / ml MTT solution, incubate at 37℃ for 4-6 hours, discard the MTT solution , and finally add 150 μl of DMSO to shake and mix well, place in a microplate reader, and detect the absorbance value at a measuring wavelength of 490 nm. The result is as figure 1 As shown, shikonin can significantly inhibit the growth of lung cancer cell A549.
experiment example 2
[0024] Experimental example 2: Shikonin up-regulates the expression level of p53 protein in lung cancer cells
[0025] A549 cells in the logarithmic growth phase were inoculated into 6-well culture plates at 6 × 104 cells / well, and cultured overnight. 1, 2 μM) of fresh culture medium for 72 hours, digested, collected cells, centrifuged, washed twice with PBS, washed off the supernatant, then added appropriate amount of RIPA and 1% protease inhibitor PMSF, lysed cells for 45 minutes, collected the supernatant The supernatant was used to determine the protein content, and the expression of p53 protein was detected by Western blotting. The result is as figure 2 As shown, shikonin can significantly up-regulate the expression level of p53 protein in A549 cells after acting on it.
experiment example 3
[0026] Experimental Example 3: Shikonin Induces Senescence of Lung Cancer Cells
[0027] A549 cells in the logarithmic growth phase were inoculated in 6-well plates at 6×104 cells / well, and cultured overnight. After the medium was cultured for 72 hours, the old medium was discarded, washed twice with PBS, fixed with 1X Fixative solution for 10 minutes, discarded, then washed twice with PBS, and then stained with β-galactosidase solution, incubate overnight, and detect the degree of cell senescence under a fluorescent inverted microscope. The more blue cells, the deeper the blue of the cells, and the more serious the degree of cell senescence. The result is as image 3 As shown, after the effect of shikonin, the degree of cell senescence increased in a dose-dependent manner.
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