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Long non-coding RNA AY927503 and application thereof

An RNA-AY927503, non-coding technology, applied in the field of genes, can solve problems such as changing integrins

Active Publication Date: 2016-11-23
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, how sulfatide alters the expression of integrin αV has not been reported so far

Method used

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  • Long non-coding RNA AY927503 and application thereof
  • Long non-coding RNA AY927503 and application thereof
  • Long non-coding RNA AY927503 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0107] Sulfatide promotes the migration of human liver cancer cells and the expression experiment of long non-coding RNA-AY927503

[0108] (1) Materials and methods

[0109] 1. Materials

[0110] 1.1 Cell culture and treatment

[0111] 1.1.1 Cell lines

[0112] The culture methods of all cell lines were carried out according to the optimal conditions suggested by ATCC (American Type Culture Collection); human liver cancer cell lines SMMC-7721, BEL-7404, Hep3B, HepG2, Huh-7 were preserved by our laboratory; CST8 (CST in Overexpression in SMMC-7721 cell line) and the corresponding empty control Mock stable transfection cell line, Chp2 / siCST (CST is disturbed in SMMC-7721 cell line) and the corresponding empty control Scramble stable transfection cell line were obtained from this experiment Cell construction and storage; LM3 cell line was donated by Shanghai Liver Cancer Research Center; L02, Hela and HEK293T cell lines were purchased from ATCC.

[0113] 1.1.2 Cell culture re...

Embodiment 2

[0185] Example 2 Long non-coding RNA-AY927503 promotes the migration of human liver cancer cells and the expression experiment of integrin αV

[0186] (1) Materials and methods

[0187] 1. Materials

[0188] 1.1 Plasmid construction and transfection

[0189] 1.1.1 Strains

[0190] Escherichia coli (E.coli.) strain Top10 is preserved by our laboratory.

[0191] 1.1.2 Plasmids and corresponding vectors

[0192] The RNA interference plasmid vector pSilencer 4.1-CMV puro was purchased from Ambion.

[0193] The overexpression plasmid vector pcDNA3.1-basic was purchased from Invitrogen.

[0194] The primers were designed by our laboratory and synthesized by Shanghai Bioengineering Technology Co., Ltd. The specific sequences are as follows:

[0195]

[0196] 1.1.3 Reagents related to plasmid construction

[0197] Restriction endonucleases HindIII, BamH I, EcoR I and Xba I, T4 DNA ligase, and common PCR kits were purchased from Dalian Bao Biological Engineering Co., Ltd. (Ta...

Embodiment 3

[0360] Example 3 Long non-coding RNA AY927503 regulates the promoter experiment of integrin αV

[0361] (1) Materials and methods

[0362] 1. Materials

[0363] 1.1 In situ hybridization

[0364] The Enhanced Sensitive ISH Detection Kit II (AP) of Wuhan Boster Bioengineering Co., Ltd. (BOSTER) was used.

[0365] Various buffers:

[0366] 3% citric acid: add citric acid (C 6 h 8 o 7 ·H 2 O) 3g, pH 2.0 or so.

[0367] 2×SSC: add 17.6g sodium chloride, trisodium citrate (C 6 h 5 o 7 Na 3 2H 2 O, molecular weight 294) 8.8g.

[0368] 0.5×SSC: Add 100mL 2×SSC to 300mL distilled water.

[0369] 0.2×SSC: Add 30mL 2×SSC to 270mL distilled water.

[0370] 20% glycerin: 20mL glycerin plus 80mL distilled water.

[0371] 0.5M TBS: 1000mL distilled water plus 30g sodium chloride, TRIS 1.2g, pure acetic acid 0.4-0.5mL, pH 7.2-7.6.

[0372] 0.01M TBS (PH9.0-9.5): Add 9g NaCl and 1.2g Tris to 1000mL distilled water.

[0373] Polylysine was purchased from Shanghai Pufei Biotec...

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Abstract

The invention belongs to technical field of genes, and relates to a long non-coding RNA AY927503 and application thereof. It is proved through experiments that the long non-coding RNA AY927503 can adjust migration of human hepatocellular carcinoma cells, sulfatide can adjust migration of the human hepatocellular carcinoma cells through the long non-coding RNA AY927503, express of integrin alpha V can be adjusted, and the long non-coding RNA AY927503 can adjust migration of the cells through the integrin alpha V; it is proved through experiments that the long non-coding RNA AY927503 is mainly located in cytoplasm and also exists in cell nuclei, the long non-coding RNA AY927503 can adjust the activity of a promoter of the integrin alpha V, and the long non-coding RNA AY927503 is not combined with a transcription factor on the promoter of the integrin alpha V but is directly combined with the promoter of the integrin alpha V. According to the long non-coding RNA AY927503 and the application thereof in gene control, the long non-coding RNA AY927503 can be directly combined with the promoter of the integrin alpha V, adjust the transcriptional activity of the integrin alpha V and then influence the behavior of the cells; the long non-coding RNA AY927503 can be used for preparing a target molecule of the human hepatocellular carcinoma cells.

Description

technical field [0001] The present invention belongs to the field of gene technology, and specifically relates to long non-coding RNA AY927503 and its use in gene regulation. The long non-coding RNA-AY927503 can directly bind to the promoter of integrin αV, regulate the transcriptional activity of integrin αV, and then affect cells behavior; especially the use of the long non-coding RNA AY927503 as a target molecule for human liver cancer cells. Background technique [0002] It is reported in the prior art that primary hepatocellular carcinoma (hepatocellular carcinoma, HCC) is the most common type of liver cancer. Liver cancer is one of the fifth most common cancers in the world and has become the third leading cause of cancer death in the world, seriously threatening human health. It has been reported that the recurrence and metastasis of primary hepatocellular carcinoma after surgical resection are the main obstacles in the treatment of liver cancer, and the study of the...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12Q1/68C12Q1/02
Inventor 吴兴中戚冰
Owner FUDAN UNIV
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