siRNA capable of inhibiting MAT2A genetic expression and application of siRNA

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

Active Publication Date: 2015-11-11
李家平 +3
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  • 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 genetic expression and application of siRNA
  • siRNA capable of inhibiting MAT2A genetic expression and application of siRNA
  • siRNA capable of inhibiting MAT2A genetic expression and application of siRNA

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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] Search in GenBank to obtain the cDNA sequence of human MAT1A, namely GeneID: 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 (Codingsequence, CDS), namely GeneID: 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 on...

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Abstract

The invention discloses siRNA capable of inhibiting MAT2A genetic expression and application of the siRNA. The positive-sense strand of siRNA MAT2A-1 capable of inhibiting the MAT2A genetic expression is shown as SEQ ID NO:2, and the antisense strand of the siRNA MAT2A-1 is shown as SEQ ID NO:3. The positive-sense strand of siRNA MAT2A-2 capable of inhibiting the MAT2A genetic expression is shown as SEQ ID NO:4, and the antisense strand of the siRNA MAT2A-2 is shown as SEQ ID NO:5. Lentivirus vector transfection liver cancer cells of lentivirus expression vector combination overexpression MAT1A genes built according to the siRNA inhibiting the MAT2A genetic expression successfully inhibit hepatocellular carcinoma angiogenesis, migration, invasion and lung metastasis, the nucleotide sequence of the MAT1A genes is shown as the 256th-144th bases of SEQ ID NO.1. The siRNA can be further applied to preparing medicine treating liver cancer diseases 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 (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 meta...

Claims

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Application Information

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