Applications of LINCR-0001 in preparation of gastric cancer diagnostic products and treatment drugs
A product, gastric cancer technology, applied in the field of biomedicine, can solve problems such as poor prognosis and achieve the effect of improving survival rate
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Embodiment 1
[0048] Example 1 Differentially expressed LncRNA research
[0049] 1. Research object
[0050] Surgical specimens (gastric cancer tissues and corresponding adjacent normal tissues) of 50 patients with primary gastric cancer who underwent radical gastrectomy in the Department of Oncology of the hospital were collected. Among the 50 patients, there were 30 males and 20 females; their age ranged from 38 to 79 years, with an average of 61 years old. None of the patients had received adjuvant therapy before operation. Immediately after RNA extraction, tissue samples were stored at -80°C.
[0051] Inclusion criteria: a. Preoperative diagnosis of primary gastric cancer, and no tumor treatment; b. No other malignant tumors; c. No other complications, such as gastric perforation, gastrointestinal bleeding, gastrointestinal obstruction, etc. The general condition of the patient was good before operation; d. No other chronic diseases, such as hypertension, diabetes, etc.
[0052] 2. D...
Embodiment 2
[0056] Example 2 inhibits the expression of LINCR-0001
[0057] 1. Cell culture and transfection
[0058] Cell culture: Gastric cancer cell line BGC-823 was used in DMEM containing 10% FBS and placed in 5% CO 2 , Saturated humidity, cultured in a 37°C carbon dioxide incubator. The culture medium was changed every two days, and the cells were digested with 0.25% trypsin when subcultured.
[0059] siRNA transfection: The day before transfection, cells are digested and inoculated into culture dishes or culture plates, and the number of cells inoculated should ensure that the density of 30-50% can be reached on the second day of transfection. siRNA transfection strictly according to Lipofectamin TM According to the 2000 specification, after 4-6 hours, replace with fresh culture medium containing 10% FBS, and continue to cultivate for 48 hours.
[0060] 2. siRNA design
[0061] Design of siRNA (small interfering RNA): siRNA sequence was designed in the specific sequence region...
Embodiment 3
[0070] Example 3 Determination of Inhibition of LINCR-0001 Expression on the Proliferative Ability of Gastric Cancer Cells
[0071] 1. Steps:
[0072] BGC-823 cells in the logarithmic growth phase were seeded on 96-well culture plates. After 12 hours of culture, the cells were transfected with siRNA-LncRNA and siRNA-NC respectively. After 4-6 hours, DMEM containing 10% FBS was replaced and cultured for 72 hours. Add 10 μL CCK8 to each well of each plate, incubate in the incubator for 2 hours, and then use a microplate reader to detect the absorbance value at a wavelength of 450 nm. Five replicate wells were set up for each group of cells, the experiment was repeated 3 times, and the cell growth curve was drawn.
[0073] 2. Statistical analysis
[0074] The experimental data were expressed as mean ± standard deviation, and SPSS15.0 statistical software was used to conduct one-way analysis of variance (ANOVA) or t test. P<0.05 means the difference is statistically significant...
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