Epigenetic drug for treating osteosarcoma
An osteosarcoma and drug technology, applied in the field of medicine, can solve the problem of not making obvious progress, and achieve the effects of inhibiting the growth and metastasis of osteosarcoma cells, reducing drug side effects, and improving anti-tumor efficacy.
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Embodiment 1
[0021] GSK-J4 is prepared according to different concentrations and ready for use.
[0022] The effects of different concentrations of GSK-J4 on the proliferation of human osteosarcoma cells 143B, MG63, MNNG, SJSA1 and U2 were tested by MTT assay. 143B (ATCC No. CRL-8303) will be in a period of exponential growth TM , human source, epithelioid adherent growth), MG63 (ATCC number CRL-1427 TM , human source, epithelioid adherent growth), MNNG (ATCC number CRL-1547 TM , human source, epithelioid adherent growth), SJSA1 (ATCC number CRL-2098 TM , human, epithelioid adherent growth) and U-2OS cells (ATCC number HTB-96 TM , human source, epithelial-like adherent growth), were digested with trypsin, blown into a single cell suspension, counted, and inoculated into a 96-well culture plate. Cultivate in the incubator for 24 hours. After the cells adhere to the wall, add different concentrations of GSK-J4 (purchased from Selleck, product number S7070), and stop the culture after 48 ...
Embodiment 2
[0027] Using animal models to verify that GSK-J4 has an inhibitory effect on the growth of osteosarcoma. In the experiment, 6-week-old female nude mice were purchased from the Guangdong Experimental Animal Center, weighing between 15 and 20 g, and were reared in separate cages under the condition of no special pathogenic bacteria (SPF), with 12 hours of alternating light and free food and drink. Osteosarcoma cells 143B and SJSA1 were cultured in DMEM high-glucose medium containing 10% high-quality fetal bovine serum until the logarithmic growth phase was used for experiments. When the cells cover the bottom of the culture flask, digest the cells with 5% trypsin, stop the digestion with high-quality fetal bovine serum, blow gently until the cells are completely detached from the wall, centrifuge at 1500 rpm for 5 minutes, and discard the supernatant. The cells were then washed twice with saline. After washing, resuspend the cells in normal saline and adjust the cell concentrat...
Embodiment 3
[0031] Transwell assay was used to verify that GSK-J4 inhibited the metastasis of osteosarcoma cells 143B and SJSA1 ( image 3 ).
[0032] Take a 24-well plate, add 24 Transwell chambers, divide into 8 groups, 4 groups for each type of cells, and each group has 3 replicate wells. Add 50 μL of BD matrigel to each small chamber in turn, add 50 μL of serum-free DMEM medium after it solidifies, and then inoculate 143B or SJSA1 cells in the logarithmic growth phase in the small chambers in turn, and each well contains 50 μL of medium containing 2×10 4 cells. GSK-J4 was added to the upper chamber of Tranwell, the concentrations from left to right were 0μM, 2.5μM, 5μM and 10μM. 500 μL of cell culture medium containing 10% fetal bovine serum was slowly added along the well walls of the 24-well plate, and cultured in a 5% CO2, 37°C constant temperature and humidity incubator for 24 hours. After the end of the experiment, remove the medium in the Transwell chamber and the well in tu...
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