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Application of 20(S)-ginsenoside-Rg3 in reversing drug resistance of glioma cells to chemotherapeutic drugs

A technology of glioma cells and ginsenosides, which can be used in drug combinations, anti-tumor drugs, medical preparations containing active ingredients, etc. Effect

Pending Publication Date: 2020-05-19
XIN HUA HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] In view of the above-mentioned shortcoming of the prior art, the object of the present invention is to provide the application of 20-(S) ginsenoside-Rg3 in reversing the drug resistance of glioma cells to chemotherapeutic drugs, for solving the problems in the prior art. The problem of tumor resistance to temozolomide

Method used

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  • Application of 20(S)-ginsenoside-Rg3 in reversing drug resistance of glioma cells to chemotherapeutic drugs
  • Application of 20(S)-ginsenoside-Rg3 in reversing drug resistance of glioma cells to chemotherapeutic drugs
  • Application of 20(S)-ginsenoside-Rg3 in reversing drug resistance of glioma cells to chemotherapeutic drugs

Examples

Experimental program
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Effect test

Embodiment 120

[0081] Example 1 Toxic effects of 20(S)-ginsenoside-Rg3 and TMZ on glioma

[0082] GBM-XX is the primary cell line derived from the surgical specimens of World Health Organization grade IV glioma patients. The resection was performed according to the protocol approved by the ethics committee of our hospital, and the patients' prior informed consent was obtained. The medium consisted of DMEM (Life Technologies / Gibco, Carlsbad, California, USA) and 10% fetal bovine serum (FBS; Life Technologies / Gibco, Carlsbad, California, USA), 100u / ml penicillin and 100 μg / ml streptomycin (Gibco, Grand Isle, NY). cells at 1 x 10 5 Cells / ml inoculated. Culture conditions: temperature 37°C, 5% CO 2 and 100% humidity.

[0083]T98G, U118 and GBM-XX cells were seeded in 96-well flat-bottom plates at 5000 cells / well, cultured in DMEM supplemented with 10% FBS, and then treated with increasing concentrations (0, 60, 120, 240, 360 and 480 μM ), 20(S) ginsenoside-Rg3, (0, 75, 150, 300, 450 and 600...

Embodiment 220

[0086] Example 2 Effect of 20(S)-ginsenoside-Rg3 on the expression of MGMT

[0087] T98G, U118 and GBM-XX were treated with 20(S)-ginsenoside-Rg3 (100 μM) for 72 hours.

[0088] mRNA assay: Total RNA was extracted from cell lines using TRIzol (TaKaRa, Dalian, China) according to the manufacturer's protocol. For mRNA analysis, RNA was reverse transcribed into cDNA using a primer-script one-step RT-PCR kit (TaKaRa, Dalian, China). The cDNA template was amplified by RT-PCR using the SYBR Premix Eraser Kit (TaKaRa, Dalian, China). GAPDH was used as an internal control. Real-time PCR was performed in triplicate. The primer sequences used are as follows: for GAPDH primers are as follows; SEQ ID NO1.: 5'-gtcaacggattggtctggtatt-3' (forward) and SEQ ID NO2.: 5'-agtctgggggcagtgat-3' (reverse); MGMT primers are as follows: SEQ ID NO3.: 5'-gttatgaatgtaggagcccttatg-3' (forward), SEQ ID NO4.: 5'-tgacaacgggaatgaagtaatg-3' (reverse). The result is as Figure 1c As shown, 20(S)-ginsenosi...

Embodiment 320

[0090] Example 3 20(S)-ginsenoside-Rg3 inhibits the expression of MGMT in glioblastoma by regulating the Wnt / β-catenin pathway

[0091] T98G, U118 and GBM-XX were treated with 20(S) ginsenoside-Rg3 (100 μM) for 72 hours, and Wnt / β-catenin pathway-related proteins were determined.

[0092] Western blot analysis Cells were washed with PBS and then lysed with RIPA lysis buffer (Solarbio, China) and protease inhibitors (Roche Applied Science, Switzerland). Protein concentration was measured using BCA protein assay kit (Beyotime Biotechnology, China). Equal amounts of proteins were subjected to 10% SDS-polyacrylamide gel electrophoresis and transferred to PVDF membranes. Membranes were subsequently blocked with 5% skim milk for 2 h and incubated with primary antibodies overnight at 4 °C. The primary antibodies used were anti-MGMT (1:500, American Abcam Company) and anti-Survivin (1:1000, American Abcam Company), anti-β-catenin, anti-CD44, anti-C-Jun, anti-C-Myc, anti-cyclinD1, an...

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Abstract

The invention provides an application of 20(S)-ginsenoside-Rg3 in reversing drug resistance of glioma cells to chemotherapeutic drugs. Researches find that 20(S)-ginsenoside-Rg3 inhibits expression ofan MGMT gene by inhibiting a Wnt / beta-catenin signal pathway and inhibiting LncRNA NKILA, significantly reverses MGMT mediated drug resistance of glioma cells to temozolomide, promotes apoptosis of tumor cells, and plays a very positive role in treatment of glioma patients.

Description

technical field [0001] The present invention relates to a new use of a chemical substance, in particular to a use of 20(S)-ginsenoside-Rg3 in preparing a drug for reversing drug resistance of glioma cells to chemotherapeutic drugs. Background technique [0002] 20(S)-ginsenoside-Rg3 (Ginsenoside Rg3), molecular formula C 42 h 72 o 13 , molecular weight 785.01g / mol, CAS number: 14197-60-5, tetracyclic triterpene saponin monomer isolated from ginseng. [0003] Glioma is the most common primary malignant brain tumor of the central nervous system in adults. Currently, surgery combined with postoperative adjuvant radiotherapy and chemotherapy is the preferred option for patients with glioma. Temozolomide (TMZ) is a first-line chemotherapy drug for patients with glioma. Despite this, the prognosis of glioma remains poor, with a median survival of only 14.6 for high-grade glioma. Temozolomide induces cell cycle arrest and apoptosis by methylating the 6th oxygen atom of guanin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/704A61K31/4188A61P35/00
CPCA61K31/704A61K31/4188A61P35/00A61K2300/00
Inventor 郑学胜陈正
Owner XIN HUA HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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