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Application of tanshinone IIA in preparation of medicament for treating p53 mutational or deficient tumor

A tumor drug, tanshinone technology, applied in the field of application of tanshinone IIA in the preparation of tumor drugs for the treatment of p53 mutations or deletions, which can solve the problems of reduced tumor sensitivity

Inactive Publication Date: 2012-07-18
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that more than 50% of human tumors have P53 gene mutations or deletions, and P53 gene mutations and deletions often lead to reduced sensitivity of tumors to chemotherapy drugs

Method used

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  • Application of tanshinone IIA in preparation of medicament for treating p53 mutational or deficient tumor
  • Application of tanshinone IIA in preparation of medicament for treating p53 mutational or deficient tumor
  • Application of tanshinone IIA in preparation of medicament for treating p53 mutational or deficient tumor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Embodiment 1.MTT experiment

[0013] Experimental Materials:

[0014] Human non-small cell lung cancer cells A549 cells and H596 cells were purchased from ATCC at 37°C, 5% CO 2 Under conventional conditions, the medium was RPMI-1640 (Gibco) containing 10% calf serum (PAA). The NQO1 gene was synthesized from the whole gene, and IRES2-EGFP was fused downstream to construct a lentiviral expression vector containing the NQO1-IRES-EGFP fragment. The 293T cells were used to package the lentivirus, the virus stock solution was concentrated, and the virus titer was determined. Wild-type H596 cells were infected to construct a H596 cell line (H596-NQO1) stably expressing NQO1. RNAiMAX transfection reagent, control siRNA, and P53 siRNA were purchased from Invitrogen. P53 antibody was purchased from Millipore.

[0015] experimental method:

[0016] 1. P53 gene silencing

[0017] A549 cells were seeded in a 96-well plate at a density of 8000 cells / well. After 18-24 hours, a...

Embodiment 2

[0021] Embodiment 2.TUNEL experiment

[0022] Experimental material: with embodiment 1.

[0023] experimental method:

[0024] A549 cells or H596 cells were seeded in 6-well plates, and when the cell density reached 85% confluence, the A549 cells were administered. The experimental groups were as follows:

[0025] (1) H596-NQO1 cell grouping includes: TSA 0μM, 10μM, 40μM administration groups;

[0026] (2) A549 cell grouping: control siRNA transfection plus blank medium group

[0027] TSA 40μM administration group after control siRNA transfection

[0028] P53siRNA transfection plus blank medium group

[0029] TSA40μM administration group after P53siRNA transfection

[0030] The administration time was 48h. ApoBrdU DNA Fragmentation Assay Kit (BD) was used for TUNEL detection. The detection steps are as follows:

[0031] Fix cells → wash cells → TdT / BrdU labeled cells → wash cells → Anti-BrdU-FITC stained cells → wash cells → PI / Rnase treatment → flow cytometric analysi...

Embodiment 3

[0032] Example 3. Mitochondrial membrane potential detection

[0033] Experimental materials: A549 cells, JC-1 dye was purchased from KGI.

[0034] experimental method:

[0035] A549 cells at 1.5×10 4 Cells / well were seeded in 24-well plates, and the experimental groups were as follows

[0036] Control siRNA transfection plus blank medium group; control siRNA transfection TSA 40μM administration group;

[0037] Add blank medium group after P53siRNA transfection; TSA40μM administration group after P53siRNA transfection

[0038] After 24 hours of administration, remove the medium, wash the cells once with Hank's, add 3 μM JC-1 reagent and incubate at 37°C for 15 minutes, wash the cells twice with Hank's, and use synergy H1 multifunctional medium to determine the JC-1 polymer The red fluorescence emitted (excitation wavelength: 530nm, emission wavelength: 590nm) and the green fluorescence emitted by JC-1 monomer (excitation wavelength: 490nm, emission wavelength: 529nm) were...

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Abstract

The invention relates to the field of natural medicaments, in particular to the application of tanshinone IIA in preparation of a medicament for treating p53 mutational or deficient tumor. The tanshinone IIA can induce cellular poison and apoptosis of p53 mutational H596-NQO1 non-small cell lung cancer cells; and simultaneously, in wild A549 cells of p53, when P53 small interfere ribose nucleic acid (siRNA) silences endogenous P53 genes, the tanshinone IIA can still induce the cellular poison and apoptosis of the A549 cells, reduction in the mitochondrial membrane potential of the A549 cells induced by the tanshinone IIA is not influenced in the process of treating the P53siRNA, the ratio of BAX / Bcl-x1 is increased, cytochrome C is released, and Caspase is activated, so that an anti-tumor effect which does not depend on the p53 is generated.

Description

technical field [0001] The invention relates to the field of natural medicines, in particular to the application of tanshinone IIA in the preparation of medicines for treating p53 mutation or deletion tumors. Background technique [0002] P53 is a tumor suppressor gene. The p53 protein encoded by it can exert anti-tumor effects through various mechanisms. When DNA suffers continuous damage, it can activate DNA repair function-related proteins and arrest the cell cycle at G1 / S period to provide sufficient time for DNA repair. When the DNA damage reaches an irreparable level, the p53 protein can initiate the apoptosis process and ensure the stability of the genome. Apoptosis mediated by p53 protein is an important mechanism for antitumor drugs to exert their efficacy. Studies have shown that more than 50% of human tumors have P53 gene mutations or deletions, and P53 gene mutations and deletions often lead to decreased sensitivity of tumors to chemotherapy drugs. Therefore, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/58A61P35/00
Inventor 王广基郝海平刘芳刘慧颖吴晓兰刘淼廖珂程学芳吴梦秋
Owner CHINA PHARM UNIV
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