A long non-coding RNA and its application in the diagnosis/treatment of gastric cancer
A non-coding, RNA-LINC00707 technology, applied in the field of genetic engineering, can solve problems affecting patient prognosis, recurrence or metastasis of gastric cancer
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Embodiment 1
[0134] Example 1 Detection of the expression of LINC00707 in tissues and cells
[0135] Take 0.1g tissue, grind it sufficiently with liquid nitrogen (into powder) or 1-5×10 7 Discard the culture medium from the cells, and rinse twice with pre-cooled PBS. Add 1ml of Trizol lysate, pipette and mix with an enzyme-free pipette tip, let stand for 5min, and transfer the lysate into a pre-labeled 1.5ml enzyme-free centrifuge tube. Centrifuge at 7500g at 4°C for 5 minutes, take the supernatant and add 1 / 5 volume of chloroform, invert and mix for 30 seconds, and let stand for 2 minutes. Centrifuge at 12000g for 15min at 4°C. The solution is divided into three layers (aqueous phase-white precipitate-red organic matter), transfer the aqueous phase layer to a new 1.5ml centrifuge tube, try not to absorb the white precipitate. Add an equal volume of isopropanol, mix gently by inversion, and let stand for 5-10min. Centrifuge at 12000g for 10min at 4°C. Discard the supernatant, add 1ml ...
Embodiment 2
[0155] Example 2 Upregulated LINC00707 is significantly associated with aggressive tumor characteristics and poor prognosis of gastric cancer
[0156] In order to further explore the important role of LINC00707 in the malignant process of gastric cancer, we divided the expression levels of LINC00707 in 60 pairs of gastric cancer tissues into high expression group (n=30) and low expression group (n=30), and analyzed the relationship between the expression level and clinical Correlation between pathological parameters and patient prognosis. Notably, high expression of LINC00707 in gastric cancer was significantly correlated with TNM stage (p=0.035) and tumor size (p=0.017) and lymph node metastasis (p=0.003). However, the expression of LINC00707 was not correlated with other parameters such as sex (p=0.795) and age (p=0.301) etc. (Table 1). In order to determine the relationship between the expression of LINC00707 and the prognosis of patients with gastric cancer, the curves of...
Embodiment 3
[0160] Example 3 Effect of LINC00707 on the proliferation of gastric cancer cells
[0161] MTT experiment: BGC-823, SGC-7901, and MGC-803 cells 24 hours after transfection were seeded in 96-well culture plates at 3000 cells per well; after 80% of the cells adhered to the wall, the cells were synchronized for 12 hours, and the original cells were discarded. There is a culture medium. Set 6 replicate wells for each sample, and the total reaction volume of each well is 200 μl; add 20 μl of MTT reaction solution (5 mg / ml, dissolved in PBS) to each well, and incubate at 37°C in the dark for 4 hours; discard the supernatant, and Add 150 μl dimethyl sulfoxide (DMSO), shake on a micro-sample shaker for 10 min, and measure the absorbance at a wavelength of 490 nm with a microplate reader. The results showed that interference with LINC00707 could significantly inhibit the proliferation of BGC-823 and SGC-7901 cells; overexpression of LINC00707 promoted the proliferation of MGC-803 ( ...
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