A detection, treatment and prognosis target and application of cholangiocarcinoma
A cholangiocarcinoma-specific technology, applied in the field of biomedicine, can solve the problems of insensitivity of cholangiocarcinoma to chemotherapy and difficulty in entering the anatomical location of cholangiocarcinoma
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[0022] Cell Culture and Transfection
[0023] Human cholangiocarcinoma cell lines QBC939 and RBE obtained from the Chinese Academy of Sciences (Shanghai) type culture cell bank were used at 37°C, 5% CO 2 1640 medium (Hyclone, Logan, UT, USA) supplemented with 10% fetal bovine serum (Hyclone, Logan, UT, USA) and penicillin / streptomycin. After entering the logarithmic growth phase, the cells were washed 3 times with PBS, digested with trypsin, and placed in 6-well plates. When the cell density of the 6-well plate reached 50-60% confluency, Lipofectamine 2000 was used to transfect siRNA (Invitrogen, USA). siRNA targeting USP8 (si-USP8) was synthesized by OriGene (Beijing, China), and the siRNA negative control (OriGene) was used as a negative control (si-NC).
[0024] qRT-PCR analysis
[0025] 24 hours after transfection with siRNA, total RNA was extracted from the cells with Ultrapure RNA Kit (CWBIO, Beijing, China), and reverse transcribed into cDNA with HiFiScript cDNA Synt...
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