Method for inhibiting stemness of pancreatic cancer stem cells with miR-200a

[0028] A method for miR-200a inhibiting stemness characteristics of pancreatic cancer stem cells, comprising the following steps:

[0029] A) Human pancreatic cancer cell line PANC-1 culture

[0030] DMEM culture medium containing 10% fetal bovine serum (FBS) was used as the medium, at 37°C, 5% CO 2 The human pancreatic cancer cell line PANC-1 was cultured under saturated humidity conditions, passaged once every 2-3 days, and digested with 0.25% trypsin every passage;

[0031] B) Detection and sorting of pancreatic cancer stem cells by flow cytometry

[0032] (1) Digest PANC-1 cancer cells into single cell suspension

[0033] After the cells grow to the logarithmic growth phase, remove the culture medium, wash once with PBS, add 0.25% trypsin, and digest at 37°C for 5 minutes; DMEM culture medium), pipette several times; transfer to a sterile centrifuge tube, centrifuge at 1000r / min for 5min, resuspend cells in PBS, and count on a hemocytometer;

[0034] (2) Antibody Incuba