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Application of lincRNA in the preparation of drugs for regulating methotrexate tumor treatment drug resistance

A technology for regulating methotrexate and tumor treatment, which can be used in anti-tumor drugs, gene therapy, drug combinations, etc., and can solve problems such as methotrexate resistance that are not well resolved

Active Publication Date: 2016-06-15
GUANGZHOU INST OF ADVANCED TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The prior art has not yet solved the problem of methotrexate drug resistance in clinical application
Moreover, there is no report in the prior art that lincRNA can regulate the drug resistance of methotrexate tumor therapy, and there is no report that non-coding RNAH19 can regulate drug resistance

Method used

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  • Application of lincRNA in the preparation of drugs for regulating methotrexate tumor treatment drug resistance
  • Application of lincRNA in the preparation of drugs for regulating methotrexate tumor treatment drug resistance
  • Application of lincRNA in the preparation of drugs for regulating methotrexate tumor treatment drug resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1MTX handles the impact on HT-29 cell morphology

[0029] use 10 -5 Colon cancer cells HT-29 were treated with MTX for 3 days, and the HT-29 cells without MTX treatment were used as the control to observe the effect of MTX on the morphology of HT-29 cells. The result is as figure 1 As shown, the colon cancer cells treated with MTX were in the form of stretched sheets or spindles ( figure 1 B), while untreated HT-29 cells are normally round ( figure 1 A).

[0030] The expression of embodiment 2H19 enhances the tolerance of MTX in HT-29 cells

[0031] HT-29 cells were divided into 2×10 4 Density seeded in 12-well plate, then 10 -6 μM and 10 -5 After treatment with MTX at μM concentration for 3 days, the cells were collected, RNA was extracted, and cDNA was obtained after reverse transcription. Then real-time fluorescent quantitative PCR technology was used to detect the expression abundance of H19 in HT-29 cells treated with two different MTX concentrat...

Embodiment 4H19

[0034] Example 4H19 overexpression enhances the tolerance of MTX in colon cancer HT-29 cells

[0035] The H19 overexpression plasmid was used to overexpress H19 in HT-29 cells. HT-29 cells according to 2×10 4 After the density was seeded in a 12-well plate, the H19 plasmid was transfected into HT-29 cells, and then 10 -5 The μM concentration of MTX was treated for 3 days, and the cell viability was detected by the method of cell proliferation (MTT method). The result is as Figure 5 As shown, H19 overexpression increased the tolerance of MTX in HT-29 cells.

[0036] In summary, the results of Examples 3 and 4 indicate that the expression of H19 regulates the tolerance of MTX in HT-29 cells.

Embodiment 5H19

[0037] The impact of embodiment 5H19 on miRNAs expression and ABCC2 / ABCG2 expression

[0038] Some miRNAs were predicted by bioinformatics to target both H19 and ABCC2 / ABCG2. The relationship between each miRNAs and specific target genes is shown in Table 1.

[0039] Table 1 Relationship between miRNAs and specific target genes

[0040]

[0041]

[0042] HT-29 cells were divided into 2×10 4 After the density was seeded in a 12-well plate, the H19 plasmid was transfected into HT-29 cells, and then 10 -5 After treatment with MTX at μM concentration for 3 days, the cells were collected, RNA was extracted, and cDNA was obtained after reverse transcription. Firstly, the expression of drug resistance genes ABCC2 and ABCG2 was detected. The result is as Figure 6 As shown, H19 overexpression promoted the high expression of these two drug resistance genes. Then adopt real-time fluorescent quantitative PCR technique to detect wherein miR-186, miR-212, miR-132, miR-19a, miR-...

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Abstract

The invention discloses the application of lincRNA in the preparation of drugs for regulating the drug resistance of methotrexate tumor treatment, and the coding sequence of the lincRNA is shown in SEQ ID No.1. Non-coding RNA H19 (its sequence is shown in SEQ ID No.1) acts as a miRNA sponge or miRNA competitor, which can bind to certain miRNAs, and these miRNAs can bind to the 3'-UTR of ABCC2 / ABCG2 genes, thereby inhibiting ABCC2 / The expression of ABCG2 gene can improve the tolerance of methotrexate in tumor cells; non-coding RNA H19 may be used as a diagnostic marker or molecular target of methotrexate in tumor treatment to improve the effect of tumor treatment.

Description

technical field [0001] The present invention belongs to the field of biomedicine. More specifically, the present invention relates to the application of a lincRNA (large intergenic non-coding RNA, intergenic long-chain non-coding RNA), in particular to the application of lincRNA in the preparation and regulation of methotrexate (Methotrexate, MTX) tumors Application of drugs in the treatment of drug resistance. Background technique [0002] Methotrexate is an anti-folate antineoplastic drug, which mainly hinders the synthesis of tumor cells by inhibiting dihydrofolate reductase, thereby inhibiting the growth and reproduction of tumor cells. As the earliest cytotoxic drug used in tumor treatment, methotrexate is effective against a variety of tumors including breast cancer, osteosarcoma, head and neck cancer, leukemia, and colon cancer. After long-term use, tumor cells will develop resistance to methotrexate, which hinders its clinical application. [0003] In order to bett...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61P35/00A61P35/02
Inventor 付卫明张锦芳梁伟铖
Owner GUANGZHOU INST OF ADVANCED TECH CHINESE ACAD OF SCI
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