Application of long-chain non-coding RNA LINC02481 in inhibition of proliferation of gastric cancer cells and promotion of apoptosis of gastric cancer cells
A LINC02481, long-chain non-coding technology, applied in the field of biomedicine, can solve the problems of patients not paying attention, neglect of gastric cancer, lack of specific clinical symptoms, etc., and achieve the effect of excellent diagnostic value
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Embodiment 1
[0027] Detection of the expression difference of LINC02481 in gastric cancer tissues and adjacent tissues
[0028] 1. Research object
[0029] 20 cases of gastric cancer tissues and 20 cases of corresponding paracancerous tissues were obtained from Qingdao Cancer Hospital. All tissue samples were reviewed and approved by the Hospital Ethics Committee, and all tissue samples were obtained from patients who signed informed consent. The gastric cancer tissue was confirmed to be gastric cancer by pathological examination (when the gastric cancer tissue was resected, the patient had not yet received chemotherapy, radiotherapy, etc.).
[0030] The specific organization is as follows
[0031] Numbering gender age installments TNM tumor type Tumor size (cm) 1 male 52 III T3N1M0 moderate‐poorly differentiated adenocarcinoma 5×5×1.5 2 Female 57 IA T1N0M0 Fundic stromal tumor 4×4×4.5 3 male 48 IIIC T4bN3M0 poorly differentia...
Embodiment 2
[0075] Real-time quantitative PCR detection of LINC02481 gene expression after knockdown
[0076] (1) Cell culture
[0077] Human gastric cancer cell MKN-45 was cultured in high-glucose DMEM medium (10% fetal bovine serum) at 37°C, 5% CO 2 Constant temperature cell incubator culture.
[0078] (2) si-RNA transfection
[0079] (1) The day before transfection, 2×10 5 MKN-45 cells were seeded in 6-well plates at 37°C, 5% CO 2 Culture in a constant temperature cell incubator for 24 hours, and transfect according to the instructions of Lipofectamine2000;
[0080] (2) The experimental groups were control group, si-NC group, si-LINC02481-1, si-LINC02481 siRNA-2, si-LINC02481-3.
[0081] 5'‐AUUUGUCGGAGGCAUUAUCAA-3' (SEQ ID NO:6)
[0082] 5'-GAUAAUGCCUCCGACAAAUAU-3' (SEQ ID NO:7)
[0083] si-LINC02481-2
[0084] 5'-GAAGGUGUUUGCUAGACAAAG-3' (SEQ ID NO:8)
[0085] 5'-UUGUCUAGCAAACACCUUCUG-3' (SEQ ID NO:9)
[0086] si-LINC02481-3
[0087] 5'-UCAUGAUAAAUCGAACAAGCA-3' (SEQ ID NO: ...
Embodiment 3
[0096] CCK-8 assay detects the effect of transfection si-LINC02481-2 on the proliferation ability of gastric cancer cell MKN-45
[0097] experimental method
[0098] (1) Put 2×10 3 MKN-45 cells were inoculated in 96-well plates, 100 μl per well, and three replicate wells were set up in each group, and cultured in a 5% CO2 incubator at 37°C;
[0099] (2) MKN-45 cells were transfected with si-NC and si-LINC02481-2 respectively. 4 hours before the end of the culture at 24h, 48h, and 72h, 10ul CCK-8 was added to each well, and after 4h incubation in a constant temperature incubator, 450nm Detect absorbance.
[0100] Experimental results
[0101] pass Figure 4 It can be seen that at 24h of transfection, there was no significant difference between the si-NC group and the si-LINC02481-2 group, while at 48 and 72h (48h: si-NC 0.649±0.035, si-LINC02481-2 0.431±0.027; 72h: si-NC 1.057±0.415, si-LINC02481-2 0.683±0.039), the OD value of MKN-45 cells transfected with si-LINC02481-2 ...
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