mir-2 small molecule and application
A small molecule inhibitor, mir-21 technology, which can be used in medical preparations containing active ingredients, organic active ingredients, organic chemistry, etc., and can solve the problems of high price and no real clinical application
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Embodiment 1
[0032] Example 1: miR-21 small molecule inhibitor inhibits glioma cells
[0033] Take 42mg of the compound 2,4-diamino-1,3-diazepine-5-carbonitrile and dissolve it in 3ml of DMSO to obtain a mother solution with a concentration of 0.1M. Inoculate U87 and LN229 glioma cells in 6-well plates, the number of cells is about 100,000-150,000. After 24 hours of culture in DMEM containing 10% FBS, add 2,4-diamino-1,3-diazepine The final concentration of cyclo-5-nitrile mother solution was 1.5, 7.5, 15, 30, and 75 μM, respectively, and after 0.5, 1, 2, 6, 24, and 48 hours of culture, real-time PCR was used to detect the expression of miR-21 and other miRNAs, and the cell cycle was paralleled and cell apoptosis detection. The specific method of real-time PCR: use the RNA extraction kit to extract the total RNA in the cells, and perform reverse transcription with AMV reverse transcriptase. The SYBR Green fluorescent dye incorporation method was used for Real-time PCR, U6 was used as the...
Embodiment 2
[0034] Example 2: miR-21 small molecule inhibitor inhibits breast cancer cells
[0035] Take 42mg of the compound (referring to the small molecule inhibitor of miR-21) 2,4-diamino-1,3-diazepine-5-carbonitrile, and dissolve it in 3ml of DMSO to obtain a mother solution with a concentration of 0.1M. Inoculate MCF-7 breast cancer cells in 6-well plates, the number of cells is about 100,000-150,000, and culture them in DMEM containing 10% FBS for 24 hours, then add 2,4-diamino-1,3-diazepine -5-Nitrile mother solution to make the final concentration of 1.5, 7.5, 15, 30, 75 μM respectively, after culturing for 0.5, 1, 2, 6, 24, and 48 hours, real-time PCR detects the expression of miR-21 and other miRNAs, and parallels the cell cycle and cell apoptosis detection. The specific method of real-time PCR: use the RNA extraction kit to extract the total RNA in the cells, and perform reverse transcription with AMV reverse transcriptase. The SYBR Green fluorescent dye incorporation method...
Embodiment 3
[0036] Example 3: miR-21 small molecule inhibitor inhibits gastric cancer cells
[0037]Take 42mg of the compound 2,4-diamino-1,3-diazepine-5-carbonitrile and dissolve it in 3ml of DMSO to obtain a mother solution with a concentration of 0.1M. Inoculate SCG7901 gastric cancer cells in 6-well plates, the number of cells is about 100,000-150,000. After culturing in DMEM containing 10% FBS for 24 hours, add 2,4-diamino-1,3-diazepine-5- The final concentrations of the nitrile mother solution were 1.5, 7.5, 15, 30, and 75 μM, respectively. After culturing for 0.5, 1, 2, 6, 24, and 48 hours, real-time PCR was used to detect the expression of miR-21 and other miRNAs, and parallel cell cycle and apoptosis death detection. The specific method of real-time PCR: use the RNA extraction kit to extract the total RNA in the cells, and perform reverse transcription with AMV reverse transcriptase. The SYBRGreen fluorescent dye incorporation method was used for Real-time PCR, U6 was used as t...
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