siRNA capable of inhibiting mat2a gene expression and its application

A gene expression and gene technology, which is applied to siRNA that inhibits the expression of MAT2A gene and its application field, to achieve high theoretical and practical value and the effect of inhibiting liver cancer angiogenesis

Active Publication Date: 2017-12-01
李家平 +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, so far, there are still few studies on the joint intervention of MAT1A and MAT2A on the tumorigenesis of liver cancer cells, especially whether the dysregulated expression of MAT gene affects the angiogenesis, invasion and metastasis of liver cancer has not been reported yet.

Method used

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  • siRNA capable of inhibiting mat2a gene expression and its application
  • siRNA capable of inhibiting mat2a gene expression and its application
  • siRNA capable of inhibiting mat2a gene expression and its application

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Embodiment 1

[0032] 1 Construction of a lentiviral vector overexpressing MAT1A

[0033] In order to study the effect of MAT1A on the human liver cancer cell line SMMC-7721, we constructed a lentiviral vector overexpressing MAT1A, and the gene expression plasmid vector was the lentiviral vector pCDH-CMV.

[0034] 1.1 Gene locus selection

[0035] The cDNA sequence of human MAT1A was searched in GenBank, namely Gene ID: 4143 (its sequence is shown in SEQ ID NO.1). Design a pair of primers, the forward primer sequence is: 5'-CCTGTGAGACAGTGTGCAAG-3', the reverse primer sequence is: 5'-ATGCACTCCTCTGTCTCGTC-3', use this primer pair to PCR amplify the human genome DNA template to obtain a gene fragment: MAT1A gene mRNA The coding sequence (Coding sequence, CDS) of Gene ID: 4143 (its sequence is shown in SEQ ID NO.1) No. 256-1443. The primer pair was synthesized by Yingwei Jieji (Shanghai) Trading Co., Ltd.

[0036] 1.2 PCR amplification of cDNA of MAT1A

[0037] (1) Add the reactants one by o...

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Abstract

The invention discloses siRNA capable of inhibiting MAT2A gene expression and application thereof. For the siRNA MAT2A-1 that inhibits the expression of the MAT2A gene of the present invention, its sense strand is shown in SEQ ID NO.2, and the antisense strand is as shown in SEQ ID NO.3; for the siRNA MAT2A-2 that inhibits the expression of the MAT2A gene, its sense strand As shown in SEQ ID NO.4, the antisense strand is shown in SEQ ID NO.5. The lentiviral expression vector for inhibiting the expression of the MAT2A gene constructed based on the siRNA that inhibits the expression of the MAT2A gene is combined with the lentivirus that overexpresses the MAT1A gene (its nucleotide sequence is shown in the 256th-1443rd base of SEQ ID NO.1) Vector transfection of liver cancer cells successfully inhibits hepatocellular carcinoma angiogenesis, migration and invasion, and lung metastasis, which can be further applied to the preparation of drugs for the treatment of liver cancer, and has high theoretical and application value.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to siRNA capable of inhibiting MAT2A gene expression and application thereof. Background technique [0002] Hepatocellular carcinoma (HCC) is one of the most common malignant tumors, ranking second in the incidence of cancer in the country, and the number of deaths due to liver cancer is about 350,000 every year. Currently, surgical resection and liver transplantation are the most effective treatments for liver cancer. However, due to the low rate of early diagnosis and the shortage of liver transplant donors, most patients lost the chance of radical surgery, and the 5-year survival rate of patients was only 10-20%. Transarterial chemoembolization (TACE) is the main treatment for unresectable liver cancer, which can promote tumor ischemic necrosis and inhibit its growth, but embolization is difficult to complete, and postoperative recurrence and metastasis are always the key ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/867A61K31/713A61P35/00A61P35/04
Inventor 李家平
Owner 李家平
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