Application of PPM-18 in inducing apoptosis of bladder cancer cells by activating intracellular reactive oxygen species
A PPM-18, 1.PPM-18 technology, applied in the field of biomedicine, can solve the problems of large trauma, recurrence, human injury and so on
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Embodiment 1
[0036] Embodiment 1, PPM-18 anti-bladder cancer activity experiment
[0037] PPM-18 anti-bladder cancer activity experiment mainly includes the following steps:
[0038] (1-1) In vitro culture of bladder cancer cells used in experiments;
[0039] (1-2) using different concentrations of PPM-18 or PPM-18 to treat the cells cultured in step (1-1) for different times;
[0040] (1-3) Counting the detection of bladder cancer cell viability, apoptosis and ROS indicators treated with different concentrations of PPM-18 or PPM-18 for different times;
[0041] (1-4) According to the results of step (1-3), draw a graph of the detection results of the anti-bladder cancer activity of PPM-18.
[0042] The specific operation is as follows:
[0043] (1-1) Bladder cancer cell lines EJ and T24 were made into a single cell suspension with culture medium containing 10% fetal bovine serum, and 10,000 cells per well were seeded into a 96-well plate with a volume of 100 μl per well, and cultured u...
Embodiment 2
[0064] Embodiment 2, PPM-18 cell safety experiment
[0065] The PPM-18 cell safety experiment mainly includes the following steps:
[0066] (2-1) culturing the normal human hepatocytes used in the experiment in vitro;
[0067] (2-2) Treating the cells cultured in step (2-1) with different concentrations of PPM-18;
[0068] (2-3) Statistically detect the viability and apoptosis index of cells treated with different concentrations of PPM-18;
[0069] (2-4) According to the result of step (2-3), draw a graph of the cell safety detection result of PPM-18.
[0070] The specific operation is as follows:
[0071] (2-1) Normal human hepatocytes were prepared into a single cell suspension with culture medium containing 10% fetal bovine serum, seeded into a 96-well plate with 10,000 cells per well, and the volume of each well was 100 μl, and cultured until the cell abundance reached 80 %-90%.
[0072] (2-2) To the cells cultured in step (2-1), add PPM-18 such as 1 μM, 5 μM, 10 μM, ...
Embodiment 3
[0079] Example 3, PPM-18 can inhibit the viability of bladder cancer cells and the formation of clones, but has no obvious inhibitory effect on the viability of normal cells
[0080] In this embodiment, the experiment of PPM-18 inhibiting the viability of bladder cancer cells mainly includes the following steps:
[0081] (3-1) Cell culture: culture bladder cancer cells and normal human liver cells used in experiments in vitro, and digest and subculture the seed plate after the cells grow to the logarithmic growth phase;
[0082] (3-2) 96-well plate: the bladder cancer cell lines EJ and T24 were made into a single cell suspension with 10% fetal calf serum culture medium, and seeded into a 96-well plate with 10,000 cells per well, with a volume of 100 μl per well , cultivated until the cell abundance reaches 80%-90% (generally cultivated for 24 hours);
[0083] (3-3) Dosing treatment: after 24 hours of culture, gently suck out the old culture medium, and treat the cultured cell...
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