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Application of ferroptosis inducer RSL3 in preparation of medicine for inhibiting glioblastoma

A glioblastoma and inducer technology, applied in the field of biomedicine, can solve problems such as GBM cell description, and achieve an effective treatment plan, strong inhibitory effect, and the effect of reducing invasiveness

Pending Publication Date: 2022-05-20
THE FIRST AFFILIATED HOSPITAL OF ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is worth noting that previous reports mainly focused on class I drugs (erastin), whereas the effects of class II drugs (RSL3) on GBM cells have not been described at the transcriptome level
In addition, biomarkers associated with ferroptosis and temozolomide resistance remain undiscovered

Method used

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  • Application of ferroptosis inducer RSL3 in preparation of medicine for inhibiting glioblastoma
  • Application of ferroptosis inducer RSL3 in preparation of medicine for inhibiting glioblastoma
  • Application of ferroptosis inducer RSL3 in preparation of medicine for inhibiting glioblastoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Cell Lines and Culture Conditions

[0022] 293T, LN18, LN229 cell lines were purchased from ATCC (Manassas, VA, USA). GBM1 cells are self-established primary human glioblastoma cell lines. Construction IDH1 mutant and wild-type glioblastoma cells were cultured in DMEM (Gibco) medium; all mediums were supplemented with 10% fetal bovine serum (Gibco) and 1% penicillin / streptomycin. All cells were kept at 37°C, 5% CO 2 cultured in a humidified incubator. Treat the cells with RSL3 (1 μM, Selleck, #S8155) or camptothecin (1 μM, CST, #13637), and observe the ferroptosis and apoptosis of the cells as figure 2 In D). All cells were routinely tested for mycoplasma contamination using MycoAlertPlus (Lonza), and cells used in experiments were negative for mycoplasma.

Embodiment 2

[0023] Example 2 Establishment of IDH1 mutant and wild-type glioblastoma cells

[0024]IDH1 mutant and wild-type glioblastoma cells were constructed, and the lentivirus expressing IDH1 wild-type or mutant was packaged in transfected psik-IDH1-flag (Addgene #66802) or psik-IDH1-r132h-flag (Addgene # 66803) into 293T cells into which the packaging plasmid was introduced. After selection by hygromycin and DOX, the cells were collected, and wild-type IDH1 and R132H mutant IDH1 were detected by western blot (wb).

Embodiment 3

[0025] Example 3 Cell Proliferation Analysis

[0026] Cell proliferation / growth was assessed using the CCK8 assay (HY-K0301, MCE) following the manufacturer's instructions. That is, cells were seeded at a density of 2000-10000 cells / 100 mL in a 96-well plate. After 24h the cells were treated with RSL3 and / or Temozolomide for 24h, 48h and 72h. Add CCK8 dye solution at designated time points, incubate at 37°C for 3-4h, and detect absorbance at 450nm.

[0027] The result is as figure 1 As shown, among them, LN18 and LN229 cell lines showed normal cell proliferation in the control group and the group added with the same amount of solvent DMSO; but in the drug treatment group, that is, the RSL3 administration group, the drug RSL3 inhibited cell proliferation over time. , and gradually kill the cells. It can be seen that compared with the control group, the cell proliferation of the RSL3 drug treatment group was significantly inhibited.

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Abstract

The invention provides an application of a ferroptosis inducer RSL3 in preparation of a medicine for inhibiting glioblastoma, and also provides a medicine for combined application of the ferroptosis inducer RSL3 and temozolomide. The invention provides a more effective treatment scheme for glioblastoma, and also provides a solution capable of remarkably inhibiting glioblastoma under the condition of temozolomide resistance.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of ferroptosis inducer RSL3 in the preparation of drugs for inhibiting glioblastoma. Background technique [0002] Gliomas are the most common primary brain tumors and can generally be classified into 4 histopathological grades (I-IV). Grade IV glioma, also known as glioblastoma multiform (GBM), is the most malignant form of glioma. The average survival time of GBM patients is about 15 months, and the 5-year survival rate is less than 5%. . Surgery, radiotherapy, and chemotherapy with alkylating agents such as Temozolomide are the main treatment methods. Due to the highly aggressive growth and treatment resistance, the treatment effect of glioblastoma is generally poor. Clinically, most glioblastoma patients (about 90%) are diagnosed as IDH1 / 2 wild-type, and generally have no history of glioma, and most of them are primary. In addition, about 10% of gliobla...

Claims

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Application Information

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IPC IPC(8): A61K31/437A61P35/00A61K31/4188
CPCA61K31/437A61P35/00A61K31/4188A61K2300/00
Inventor 杨飞城王岩覃艳汪川毛敏盖曲倞姚晓雪何江
Owner THE FIRST AFFILIATED HOSPITAL OF ARMY MEDICAL UNIV
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