Long-chain non-coding RNA gene marker for detecting liver cancer and application of long-chain non-coding RNA gene marker for detecting liver cancer
A long-chain non-coding, liver cancer technology, applied in the field of detecting long-chain non-coding RNA gene markers of liver cancer, can solve problems such as lack of function, RNA formation protein, etc., and achieve the effect of improving survival rate and excellent diagnostic value.
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Embodiment 1
[0028] Detection of the expression of LINC01141 in liver cancer tissues and adjacent tissues
[0029] 1. Research object
[0030] A total of 20 liver cancer tissues and corresponding paracancerous tissues were selected for the experiment. The tissues were obtained from Qingdao Cancer Hospital. All tissue samples were reviewed and approved by the Hospital Ethics Committee, and all patients who obtained the tissue samples signed informed consent. The liver cancer tissue was confirmed to be liver cancer by pathological examination (when the liver cancer tissue was resected, the patient had not yet received chemotherapy, radiotherapy and other treatments).
[0031] The specific specimens are as follows:
[0032]
[0033]
[0034]
[0035] 2. Tissue RNA Extraction
[0036] (1) Take out the tissue sample from the -80°C refrigerator, take about 50 mg of the tissue sample and put it into a mortar, add a small amount of liquid nitrogen, and grind it quickly. After the tissue...
Embodiment 2
[0067] Knockdown of the LINC01141 gene
[0068] 1. siRNA design
[0069] si-RNA-1
[0070] 5'-AUUUUGCAGAUCUAUCAUCCU-3' (SEQ ID NO.6)
[0071] 5'-GAUGAUAGAUCUGCAAAAUUC-3' (SEQ ID NO.7)
[0072] si-RNA-2
[0073] 5'-AUCAAGUAUCUAUAUGUGCCA-3' (SEQ ID NO.8)
[0074] 5'-GCACAUAUAGAUACUUGAUAC-3' (SEQ ID NO.9)
[0075] si-RNA-1 and si-RNA-2 were synthesized by Shanghai Gemma Company, and si-NC was provided by the company.
[0076] 2. Cell Culture
[0077] Human liver cancer cells HepG2 were cultured in high-glucose DMEM medium (10% fetal bovine serum), culture conditions: cell constant temperature incubator at 37°C, 5% CO 2 .
[0078] 3. siRNA transfection
[0079] (1) The day before transfection, 2×10 5 Seed HepG2 cells onto 6-well cell culture plates at 37 °C, 5% CO 2 After culturing in the incubator for 24 hours, the cells were transfected according to the instructions of Lipofectamine 2000;
[0080] (2) Experimental groups were divided into control group (empty transfe...
Embodiment 3
[0090] 1. Cell Proliferation Experiment
[0091] (1) Divide 5×10 3 HepG2 cells transfected with si-NC and si-RNA-1 were inoculated in a 96-well plate, 90 μl per well, with 3 replicate wells for each group, and placed at 37°C, 5% CO 2 cultured in a cell culture incubator;
[0092] (2) Detection was performed at 24h, 48h, and 72h respectively. During detection, 10ul of CCK-8 detection solution was added to each well, and then placed in a 37°C, 5% CO2 incubator for 4 hours;
[0093] (3) Place the cell culture plate in a microplate reader, detect the absorbance at 450 nm, and draw the growth curve.
[0094] 2. Results
[0095] pass Figure 4 It can be seen that after the experimental group was transfected with si-RNA-1, compared with the si-NC group, the proliferation of HepG2 cells was inhibited, and the proliferation rate was significantly reduced. At 72h, the OD value of the si-NC group was 1.09± 0.049, and the OD value of the si-RNA-1 group was 0.766±0.041, the difference...
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