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ASO for targeting long-chain non-coding RNA DDX11-AS1 and kit and application thereof in treatment of liver cancer

A technology of RNADDX11-AS1 and DDX11-AS1, which is applied in the field of ASO targeting long-chain non-coding RNA DDX11-AS1, and the application field of liver cancer treatment, can solve the problems of scarcity and limitation of specific targets, and achieve the reduction of expression level, Broad application prospects, the effect of inhibiting proliferation

Inactive Publication Date: 2019-11-15
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the drugs for targeted molecular therapy on the market are very limited at present. The main reason is that the known specific targets are scarce, and there is an urgent need to find new and effective drug targets.

Method used

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  • ASO for targeting long-chain non-coding RNA DDX11-AS1 and kit and application thereof in treatment of liver cancer
  • ASO for targeting long-chain non-coding RNA DDX11-AS1 and kit and application thereof in treatment of liver cancer
  • ASO for targeting long-chain non-coding RNA DDX11-AS1 and kit and application thereof in treatment of liver cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Screening of lncRNA associated with liver cancer

[0036] 1. Sample collection: 38 cases of clinically diagnosed liver cancer and paired liver cancer tissue samples were taken. These samples were surgical resection specimens of patients with liver cancer. The samples were all from the First Affiliated Hospital of Zhejiang University School of Medicine. All the above samples were obtained with the approval of the institutional ethics committee of the hospital. Clinical data of tissue samples, including clinical characteristics such as tumor size, capsule, lung metastasis, TNM stage, distant metastasis, histopathological grade, postoperative disease-free survival time and survival time.

[0037] 2. Extraction of total RNA from hepatocellular carcinoma and paired hepatocarcinoma tissues

[0038]Take 100mg of frozen tissue and grind it quickly and fully in liquid nitrogen, add 1mL Trizol and about 1 / 5 volume of chloroform, turn it upside down for 2min, let stand...

Embodiment 2

[0043] Example 2: Expression of DDX11-AS1 in liver cancer cell lines

[0044] 1. Cell culture

[0045] The routine culture of SMMC-7721 cell line adopts 1640 cell culture medium containing 10% inactivated fetal bovine serum and 1% double antibody (penicillin-streptomycin); the routine culture of Huh7 cell line adopts the medium containing 10% inactivated fetal bovine serum and 1 % double antibody (penicillin-streptomycin) in DMEM high glucose medium. and placed at 37°C, 5% CO 2 , CO under saturated humidity conditions 2 cultured in an incubator. When the cells are confluent to about 80% of the culture dish, they are subcultured, the original culture medium is sucked out, and an appropriate amount of trypsin (containing phenol red) solution is added to digest the cells. Observe the state of the cells under an inverted microscope. When the cells are round (cytoplasm retracts), aspirate the trypsin, add 1ml of RPMI1640 or DMEM high-glucose complete medium to stop digestion, r...

Embodiment 3

[0052] Example 3: The effect of ASO on the function of liver cancer cells after inhibiting the expression of DDX11-AS1

[0053] 1. Materials

[0054] Cells: Human liver cancer cell lines SMMC-7721 and Huh7 were purchased from the Cell Resource Center of Shanghai Institute of Biological Sciences, Chinese Academy of Sciences.

[0055] Reagents: Lipofectamine 2000, a transfection reagent, was purchased from Thermo Fisher Scientific, and CCK8 and PI cell cycle detection kits were purchased from MULTISCIENCES, China.

[0056] 2, cell culture is the same as embodiment 2

[0057] 3. Design and synthesis of ASO sequence targeting DDX11-AS1

[0058] The DDX11-AS1 gene sequence (NR-) and mfold software were obtained from the NCBI database, and specific targeting ASOs were designed according to the secondary structure of the long-chain non-coding RNA DDX11-AS1, and three of them were selected for synthesis and made into an ASO mixture (equal molar ratio), the specific sequence of ASO ...

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Abstract

The invention discloses an ASO for targeting long-chain non-coding RNA DDX11-AS1 and a kit and application thereof in treatment of liver cancer. By designing and synthesizing the ASO for targeting theDDX11-AS1, transfecting the ASO into a liver cancer cell line and detecting the proliferation, cell growth cycle, migration ability and invasion ability of liver cancer cells, it is confirmed that the ASO can significantly affect the occurrence and development of liver cancer by inhibiting the expression of the DDX11-AS1. A new drug development direction and target are provided for the treatmentof liver cancer.

Description

technical field [0001] The invention belongs to the field of biomedicine, and specifically relates to an ASO targeting long-chain non-coding RNA DDX11-AS1, a kit and its application in the treatment of liver cancer. Background technique [0002] Long non-coding RNA (lncRNA) is a non-coding RNA with a length greater than 200 bp, mainly transcribed by RNA polymerase II, and lacking a distinct open reading frame. lncRNAs may participate in the regulation of gene expression at the epigenetic, transcriptional and post-transcriptional levels. lncRNAs regulate pathophysiological processes through mechanisms such as gene imprinting, histone modification, chromatin remodeling, transcriptional activation, transcriptional interference, nuclear translocation, and cell cycle regulation. The recently proposed regulatory network of competing endogenous RNAs (ceRNAs) also suggests that lncRNAs may act as sponges to competitively bind to microRNAs (miRNAs), thereby inhibiting their function...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12Q1/6886A61K31/7088A61P35/00
CPCA61K31/7088A61P35/00C12N15/113C12N2310/14C12Q1/6886C12Q2600/158C12Q2600/178
Inventor 杨忠丽赵忻艺徐梦详李明定
Owner ZHEJIANG UNIV
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