Medicinal use of a kind of lycorine and bortezomib composition
A technology of bortezomib and lycorine, which is applied in the field of new pharmaceutical uses of lycorine and bortezomib compositions, can solve problems such as drug resistance, and achieve the effect of improving sensitivity, inhibiting cell autophagy, and improving cell proliferation. Effect
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Embodiment 1
[0019] Example 1: In this example, the CCK-8 method was used to detect the effects of lycorine hydrochloride and bortezomib on the survival rate of MM cells after single and combined use.
[0020] Experimental steps:
[0021] (1) Collect ARH-77 and ANBL-6 cells in the logarithmic growth phase, adjust the concentration of the cell suspension, add 100 μL to each well, and plate on a 96-well plate to make the cell density to be tested 2×10 5 a / mL;
[0022] (2) Set up a zero-adjustment group without adding cells and only medium, and a DMSO control group without adding drugs at an appropriate concentration, and set 3-5 parallel wells for each concentration;
[0023] (3) Add 10 μM lycorine hydrochloride, 15 nM bortezomib and 10 μM lycorine hydrochloride and 15 nM bortezomib respectively, and place in 5% CO 2 , incubate in a cell culture incubator at 37°C for 24 hours;
[0024] (4) Add 10 μL of CCK-8 solution to each well, incubate at 37°C in the dark for 2 hours, measure the OD v...
Embodiment 2
[0026] Example 2: In this example, the CCK-8 method was used to detect the synergistic effect on MM cells after combined use of different concentration ratios of lycorine hydrochloride and bortezomib on MM cells.
[0027] Experimental steps:
[0028] (1) Collect ARH-77 and ANBL-6 cells in the logarithmic growth phase, adjust the concentration of the cell suspension, add 100 μL to each well, and plate on a 96-well plate to make the cell density to be tested 2×10 5 a / mL;
[0029] (2) Set up a zero-adjustment group without adding cells and only medium, and a DMSO control group without adding drugs at an appropriate concentration, and set 3-5 parallel wells for each concentration;
[0030] (3) Add different concentration ratios of lycorine hydrochloride and bortezomib (2.5μM+5nM, 2.5μM+10nM, 2.5μM+15nM, 5μM+5nM, 5μM+10nM, 5μM+15nM, 10μM+5nM , 10μM+10nM or 10μM+15nM), placed in 5% CO 2 , incubate in a 37°C cell culture incubator for 24 hours;
[0031] (4) Add 10 μL of CCK-8 sol...
Embodiment 3
[0033] Example 3: In this example, the Western Blot method was used to detect the effects of lycorine hydrochloride and bortezomib on the autophagy-related proteins LC3B and Beclin1 after single and combined use on ARH-77 and ANBL-6 cells.
[0034] Experimental steps:
[0035] (1) Collect ARH-77 and ANBL-6 cells in logarithmic growth phase, adjust the concentration of cell suspension to 5×10 5 cells / mL in a cell culture flask with a total volume of 10 mL, and then add 10 μM lycorine hydrochloride, 15 nM bortezomib, and 10 μM lycorine hydrochloride and 15 nM bortezomib in combination, and use the corresponding concentration of DMSO as a control group. Place in a cell culture incubator and incubate for 24 hours;
[0036] (2) Collect the cells, wash twice with 5mL PBS, add an appropriate volume of RIPA cell lysate (containing 0.5% protease inhibitor cocktail) to each tube, mix well, lyse on ice for 30 minutes, sonicate on ice for 30 seconds, 4°C Centrifuge for 15 minutes, aspir...
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