Applications of lncRNA-RP11-290F20.3 and small interfering RNA of lncRNA-RP11-290F20.3
A small interference, gastric cancer technology, applied in the field of biomedicine, can solve the problems of incomplete clear pathogenesis of gastric cancer, unsatisfactory results in the prevention, diagnosis and treatment of gastric cancer, etc. Effect
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Embodiment 1
[0065] Detection of lncRNA-RP11-290F20.3 expression in clinical gastric cancer tissues and gastric cancer cell lines
[0066] In this example, the clinical gastric cancer tissues and their paracancerous tissues used were freshly isolated tumor tissues from Qingdao Cancer Hospital, which were cut into small tissue pieces and frozen at -80°C for later use.
[0067] In this example, the normal gastric mucosal epithelial cells and gastric cancer cell lines used were cultured and frozen under normal normal culture conditions in the laboratory.
[0068] The total RNA of gastric cancer and adjacent tissues was extracted by Trizol reagent, and the expression level of lncRNA-RP11-290F20.3 was detected by real-time fluorescent quantitative PCR.
[0069] lncRNA-RP11-290F20.3 was amplified by PCR, and the amplified nucleotide sequence was 539bp in full length, and the sequence was shown in SEQID No.2.
[0070] The PCR reaction system is calculated as 50 μL,
[0071] The PCR conditions a...
Embodiment 2
[0097] Small interfering RNA of lncRNA-RP11-290F20.3 inhibits the proliferation of gastric cancer cells
[0098] In this example, the sequence in the small interfering RNA synthesized against lncRNA-RP11-290F20.3 is
[0099] 5'-GGAGAAUAAACCCUCGGAU-3' (SEQ ID NO. 1). The transfection treatment was carried out by lipofectamine2000, and a blank group, a control group and a small interfering RNA experimental group were respectively set up. The transfection operation was carried out according to the operating instructions of the kit, and the liposome lipofectamine2000 was purchased from Invitrogen;
[0100] The EdU solution in this example was purchased from Guangzhou Ruibo;
[0101] The cell fixative in this embodiment is the PBS buffer containing 4% paraformaldehyde;
[0102] The osmotic agent in this embodiment is the PBS buffer solution of 0.5% TritonX-100;
[0103] Using the gastric cancer cell line BGC823 in Example 1 as a cell model, the gastric cancer cell line BGC823 (...
Embodiment 3
[0108] Small interfering RNA of lncRNA-RP11-290F20.3 inhibits the invasion and migration of gastric cancer cells
[0109] The gastric cancer cell line BGC823 treated with the small interfering RNA of lncRNA-RP11-290F20.3 in Example 2 was used as a cell model. The invasion ability of cells was detected by Transwell method, and the specific operation was as follows:
[0110] Coat the upper surface of the bottom membrane of the Transwell chamber with 50 mg / L Matrigel 1:10 dilution, and incubate at 37°C; digest the cells 12 hours after the small interfering RNA of lncRNA-RP11-290F20.3 is treated, and use PBS and serum-free The culture medium was washed once successively, the cells were suspended with serum-free medium, counted, and the concentration was adjusted to 3×10 5 / mL; add 600-800 μL medium containing 20% serum to the lower chamber of the Transwell (i.e. the bottom of the 24-well plate), add 100-150 μL cell suspension to the upper chamber, and continue to culture in the...
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