Application of miR-1255 in preparation of drugs for treating carcinoma of urinary bladder
A mir-1255, coding gene technology, applied in gene therapy, drug combination, antineoplastic drugs, etc., can solve the problem of lack of effective drugs for the treatment of metastatic bladder cancer
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Embodiment 1
[0031] Example 1. Acquisition of metastatic bladder cancer cell lines stably and highly expressing miR-1255
[0032] 1. Acquisition of bladder cancer cell lines
[0033] For the colorectal cancer cell line SW780 (ATCC), the culture medium is DMEM medium (Invitrogen, 31053036) containing 10% fetal bovine serum (Hyclone) and 0.2% penicillin and streptomycin (Invitrogen).
[0035] (1) Take out the freezing tube of SW780 (ATCC), immediately put it in a water bath at 37°C-40°C, and shake it quickly until it melts completely.
[0036] (2) Transfer the cell suspension into 10ml cold PBS, put it into a 15ml centrifuge tube, centrifuge the cells with a horizontal centrifuge (1000 rpm, 8 minutes) and discard the supernatant.
[0037] (3) Suspend the cells with an appropriate amount of fresh medium, transfer the cell suspension into a culture bottle, and culture in a carbon dioxide incubator at 37°C.
[0038] B. Cell culture
[0039] Put the bladder cancer c...
Embodiment 2
[0065] Example 2. Functional study of stable and high expression of miR-1255 in SW780-high cell line
[0066] 1. Cell growth curve
[0067] (1) Take the SW780-high obtained in Example 1 in a good growth state 24 hours after transfection, and digest it with a general passage method to make a cell suspension. Counting, accurately planting cells in a 96-well plate, the total number of cells per well is required to be consistent (2000 / well), the amount of culture medium added should also be consistent, and a blank control (that is, the same volume of medium) is set, each well The final volume is 100 μL. Set at 37°C, 5% CO 2 Incubate for 24 hours in the incubator.
[0068] (2) Add 10 ul of CCK-8 solution (purchased from Japan Dojin Chemical Research Institute, article number: CK04) to each well. The amount of CCK-8 solution can be increased or decreased according to specific conditions.
[0069] (3) Normal culture conditions (37°C, 5% CO 2 incubator) for 4 hours (the incubati...
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