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ShRNA for promoting apoptosis of imatinib-resistant chronic myeloid leukemia cell K562/G01 and application thereof

A technology for leukemia cells and chronic nephritis, applied in the field of new molecular targets, can solve the problems of unclear therapeutic value of imatinib-resistant CML and achieve the effect of promoting apoptosis

Active Publication Date: 2021-10-22
THE SECOND AFFILIATED HOSPITAL TO NANCHANG UNIV
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  • Description
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Problems solved by technology

But the therapeutic value of DUSP21 in imatinib-resistant CML is still unclear

Method used

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  • ShRNA for promoting apoptosis of imatinib-resistant chronic myeloid leukemia cell K562/G01 and application thereof
  • ShRNA for promoting apoptosis of imatinib-resistant chronic myeloid leukemia cell K562/G01 and application thereof
  • ShRNA for promoting apoptosis of imatinib-resistant chronic myeloid leukemia cell K562/G01 and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] 1. Differential analysis of gene expression between imatinib-sensitive and drug-resistant cells and verification of target genes

[0023] Total RNA was extracted from K562 cells sensitive to imatinib and K562 / G01 cells resistant to imatinib, and Affymetrix gene expression profile chip was used to analyze the gene expression of the two cells (see attached figure 1 ), RT-PCR verified the expression of target gene DUSP21 in different cells (see attached figure 2 ).

[0024] 2. shRNA sequence information

[0025] According to the principle of RNA interference sequence design, use DUSP21 gene as a template to design 19-21nt RNA interference target sequence, design shRNA interference sequence for the target sequence, and add appropriate restriction endonuclease sites at both ends.

[0026] Table 1. shRNA sequence information

[0027]

[0028] AgeI restriction site: ACCGGT; EcoRI restriction site: GAATTC

[0029] 3. Vector construction and lentiviral packaging

[0030...

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Abstract

The invention discloses a novel molecular target DUSP21 for promoting apoptosis of imatinib-resistant chronic myeloid leukemia cells (K562 / G01), and correspondingly provides shRNA (short hairpin ribonucleic acid) for inhibiting target molecules and application of the shRNA. The shRNA is psc60342 or psc60343, is a targeted DUSP21 gene and interferes with the expression of DUSP21, can inhibit the expression of DUSP21 in K562 / G01 and promote the apoptosis of K562 / G01 cells, can be applied to preparation of drugs for inhibiting the proliferation of human imatinib-resistant chronic myeloid leukemia cells and promoting the apoptosis, and a new way is provided for the development of new drugs for treating imatinib-resistant chronic myeloid leukemia.

Description

technical field [0001] The present invention relates to a new molecular target for promoting apoptosis of imatinib-resistant chronic myeloid leukemia cells—DUSP21, and accordingly provides a shRNA for inhibiting the target molecule and its application. It can be applied to the preparation of drugs for inhibiting the proliferation and apoptosis of human imatinib-resistant chronic myeloid leukemia cells, and provides a new way for the development of new drugs for treating imatinib-resistant chronic myeloid leukemia. Background technique [0002] Chronic myeloid leukemia (CML) is a common hematological malignancy, and its cytogenetic marker is t(9;22)(q34;q11) to form the Philadelphia chromosome (Ph), resulting in fusion BCR / ABL and encodes an oncoprotein BCR-ABL1 with strong tyrosine kinase activity. Imatinib (Imatinib mesylate, IM), as a tyrosine kinase inhibitor (TKI), can effectively reduce the level of BCR-ABL1 and improve the survival rate of patients, and is recommende...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113A61K31/713A61P35/02
CPCC12N15/1137A61P35/02C12N2310/14C12Y301/03016C12Y301/03048C12N2310/531
Inventor 黄波章海斌王小中邹叶青李书琪刘静
Owner THE SECOND AFFILIATED HOSPITAL TO NANCHANG UNIV
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