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Application of PGC-1[alpha] specific RNA interfering adenovirus

A technology of RNA interference and PGC-1, which is applied in double-stranded DNA viruses, applications, viruses, etc., can solve the problems of less research on PGC-1α and unclear effects, etc., to increase survival time, prolong survival time, and enhance therapeutic effects Effect

Active Publication Date: 2018-04-20
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, PGC-1α has been very little studied in leukemia and its role is still unclear

Method used

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  • Application of PGC-1[alpha] specific RNA interfering adenovirus
  • Application of PGC-1[alpha] specific RNA interfering adenovirus

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Embodiment 1

[0027] 1. According to the mRNA sequence of the PGC-1α gene (NM_013261) provided by Genebank, a pair of primers were designed and synthesized using the RNAi design program of Invitrogen, USA, for the preparation of PGC-1a-specific shRNA. The primer sequences are as follows:

[0028] Upstream primers:

[0029] CCGGGACTATTGCCAGTCAATTAATCTCGAGATTAATTGACTGGCAATAGTCTTTTTG,

[0030] Downstream primers:

[0031] AATTCAAAAAGACTATTGCCAGTCAATTAATCTCGAGATTAATTGACTGGCAATAGTC.

[0032] After annealing to form double-stranded DNA (according to the conventional 50 μL annealing system, including 5 μL upstream primer, 5 μL downstream primer, 5 μL commercial 10x NEB buffer, 35 μL ddH 2 0; naturally cool to room temperature in boiling water bath for 5 minutes to complete the annealing), under the action of commercial T4DNA ligase (NEB), it is connected with the sticky end of the linearized shuttle vector pENTRTM / U6 (Addgene); the ligation product is transformed into Ecoli .DH5a competent cells...

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Abstract

The invention discloses an application of PGC-1[alpha] specific RNA interfering adenovirus in reducing leukemia cell drug resistance. The PGC-1[alpha] specific RNA interfering adenovirus provided by the invention can enhance the sensitivity of acute myeloid leukemia cells to conventional chemotherapeutic drugs (an apoptotic cell proportion is increased from a range of 22.7-27.7% to a range from 40.3-50.5%), so that the survival time of a disease animal model can be obviously prolonged; taking a leukemia mouse model, which is prepared by virtue of OCI-AML3 cells, as an example, the average survival time of a leukemia mouse, under the circumstance of accepting conventional chemotherapy, is 37.9 days; and on the basis of the conventional chemotherapy, and in combination with PGC-1[alpha] specific RNA interfering adenovirus treatment, the average survival time of the mouse exceeds 45 days, which is prolonged by more than 20%.

Description

(1) Technical field [0001] The invention relates to the application of a PGC-1α-specific RNA interference adenovirus, in particular to the application in the preparation of medicines for treating acute myeloid leukemia. (2) Background technology [0002] The tumor microenvironment is the internal environment in which the tumor occurs and develops, and is composed of tumor cells themselves, interstitial cells, microvessels, interstitial fluid, and a small amount of infiltrating cells. The current research on tumor microenvironment and tumor malignant transformation mainly focuses on the following three aspects: (1) the bidirectional and dynamic relationship between tumor cells and their microenvironment; (2) how tumor cells interact with various types of cells in surrounding tissues Promote tumor invasion and metastasis; (3) How the tumor microenvironment mediates drug resistance of tumor cells. The above three aspects are both different and related. Looking back at the rese...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/713A61K35/761A61P35/02C12N15/861
CPCA61K31/713A61K35/761C12N15/86C12N2710/10343
Inventor 陈烨孟文芳胡晓佳
Owner ZHEJIANG UNIV
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