Papillary thyroid carcinoma biomarker and application thereof
A biomarker, papillary cancer technology, applied in the field of biotechnology and oncology, can solve the problem that the mechanism of PIM1 is not fully elucidated, and achieve the effect of improving efficacy and accuracy
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Embodiment 1
[0055] Example 1: Cell Culture
[0056] Nthy-Ori-3 cells (normal thyroid cell line), BCPAP cells (mutant papillary thyroid cancer cells) and TPC-1 cells (wild-type papillary thyroid cancer cells) were purchased from Cell Center, Shanghai Institute of Biological Sciences, Chinese Academy of Sciences resource. Nthy-Ori-3 and BCPAP cell lines were placed in RPMI-1640 (Gibco, U.S.) medium, 10% fetal bovine serum (Gibco, Carlsad, CA) was added to the culture medium, and placed in 37°C, 5% CO 2 Incubate in a humidified incubator.
Embodiment 2
[0057] Example 2: Transfection of small interfering RNA
[0058] The NOX4 and PIM1 siRNA sequences and their negative controls (Jiangsu, China, Biomaike Biotechnology Co., Ltd.) were designed and obtained, as shown below. use 3000 Transfection Kit (Thermo Fisher Science) was incubated for 6 h, and then mixed with respective siRNA oligonucleotides or corresponding negative controls. After transfection, place all cells in 5% CO 2 Incubate for 48 hours in a 37°C incubator.
[0059] NOX4 siRNA sequence:
[0060] Positive: 5'-(CAUCUGUUCUUAACCUCAA)dTdT-3',
[0061] Anti: 5'-(UUGAGGUUAAGAACAGAUG)dTdT-3';
[0062] PIM1 siRNA sequence:
[0063] Positive: 5'-(CAAGAUCUCUUCGACUUCA)dTdT-3',
[0064] Reverse: 5'-(UGAAGUCGAAGAGAUCUUG)dTdT-3'.
Embodiment 3
[0065] Embodiment 3: SGI-1776 treatment
[0066] SGI-1776 (Selleckchem) was dissolved in DMSO to prepare a SGI-1776 solution with a concentration of 5 mmol / L, and the prepared SGI-1776 solution was stored in a -80°C refrigerator for future use.
[0067] BCPAP and TPC1 cells were treated with SGI-1776 solutions at a final concentration of 2.5 μM and 5 μM for 24 hours, 48 hours and 72 hours, respectively, and SGI-1776 with a final concentration of 0 μM was used as a control group. In the experiment, the highest concentration of dimethyl sulfoxide in the medium that has no significant effect on cell viability is 0.05%.
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