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Application of ganodermanondiol using endonuclear cathepsin L as target spot in pharmacy

A kind of Ganoderma lucidum ketone diol, technology for preparing medicine, applied in the field of medicine

Active Publication Date: 2018-10-16
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many years ago, there was a study using the MTT method to study the anti-colorectal cancer activity of ganoderma lucidum ketonediol on gene microsatellite stable (MSS) colon cancer cells SW480 and SW1116, but no obvious activity of ganoderma lucidum ketonediol was found

Method used

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  • Application of ganodermanondiol using endonuclear cathepsin L as target spot in pharmacy
  • Application of ganodermanondiol using endonuclear cathepsin L as target spot in pharmacy
  • Application of ganodermanondiol using endonuclear cathepsin L as target spot in pharmacy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Anti-colorectal cancer activity experiment and cytotoxicity experiment based on cell growth detection

[0030] 1.1 Subjects: colorectal cancer (HCT116 cell line, Caco2 cell line), breast cancer (MCF-7 cell line), lung cancer (HLC-1 cell line), liver cancer (HepG2 cell line), gastric cancer (HGC-27 cell line) , prostate cancer (LNCaP cell line), uterine cancer (Hela cell line) and leukemia (HL60 cell line), 4 kinds of human normal cells (normal skin fibroblast NHDF, normal umbilical cord cell HUC-F2, normal lung fibroblast TIG-1 , lung fibroblasts HF19).

[0031] 1.2 Experimental drug: Ganoderma lucidum ketonediol (purchased from Wuhan ChemFaces Co., Ltd., Hubei Province, HPLC≥98%).

[0032] 1.3 Experimental reagents and equipment: CCK-8 kit (purchased from Japan Tongren Chemical), 5-fluorouracil (5-Fu, purchased from InvivoGen, USA), super microplate reader (MD, Flexstation 3).

[0033] 1.4 Experimental method: all kinds of cells in 1×10 4 Inoculate in a 9...

Embodiment 2

[0035] Example 2: Effect experiment on colorectal cancer cell apoptosis and cell cycle:

[0036] 2.1 Experimental object: colorectal cancer HCT116 cell line.

[0037] 2.2 Experimental drugs: Ganoderma ketonediol (purchased from Wuhan ChemFaces Co., Ltd., Hubei Province, HPLC ≥ 98%), Ganoderma ketone (purchased from Wuhan ChemFaces Co., Ltd., Hubei Province, HPLC ≥ 98%), thymidine (purchased from Nanjing Senbega Biotechnology Co., Ltd.).

[0038] 2.3 Experimental reagents and equipment: Fluorescent dye PI (propidium iodide, purchased from Shenzhen Leiber Biotechnology Co., Ltd.), cell apoptosis annexin V-FITC kit (purchased from British abcam Co., Ltd.), flow cytometer SH800 ( Japan Sony).

[0039] 2.4 Experimental method: colorectal cancer cells HCT116 were treated with 3×10 5 Cells / well were inoculated in a 6-well plate, and after culturing overnight to grow into a monolayer, the culture medium containing 10% fetal bovine serum was replaced with the culture medium containi...

Embodiment 3

[0041] Embodiment 3: gene chip analysis experiment:

[0042] 3.1 Experimental object: colorectal cancer HCT116 cell line.

[0043] 3.2 Experimental drugs: Ganoderma ketonediol (purchased from Wuhan ChemFaces Co., Ltd., Hubei Province, HPLC≥98%), Ganoderma ketone (purchased from Wuhan ChemFaces Co., Ltd., Hubei Province, HPLC≥98%).

[0044] 3.3 Experimental reagents and equipment: Agilent human expression profile gene chip SurePrint G3Human GEMicroarray 8x60Kv2.0 (purchased from Agilent, USA), RNA extraction kit (purchased from InvivoGen, USA).

[0045] 3.4 Experimental method: HCT116 cells were first inoculated in a 10 cm diameter petri dish and cultured until the cell monolayer covered 90% of the surface, then 100 μM Ganoderma ketone diol (GMD) was added, and the solvent DMSO solvent control group and Ganoderma ketone (GMT) were set at the same time. ) negative control group. After culturing at 37°C for 48 hours, the cells were collected and RNA was extracted, and then huma...

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Abstract

The invention discloses application of ganodermanondiol using endonuclear cathepsin L as a target spot in pharmacy. Through in-vitro cancer cell proliferation resistance experiments and cell death andcell cycle experiments of the ganodermanondiol, it is proven that the ganodermanondiol reduces a truncated CUX1 factor (active form) by inhibiting the colorectal cancer endonuclear CL activity, thenexpression of Cyclin E2 is reduced, finally, colorectal cancer cells stay in an S phase of the cell cycle, and thus the effect of inhibiting cancer cell growth is achieved. Therefore, the ganodermanondiol can be used for preventing or curing colorectal cancer-related diseases including rectal cancer, colon cancer and related diseases, caused by rectal cancer or colon cancer, of constipation, hematochezia, abdominal pain, abdominal cramps, intestinal cancer cell infiltration and migration, and the like, and thus the ganodermanondiol is suitable for being popularized and applied.

Description

field of invention [0001] The invention belongs to the field of medicine, and in particular relates to the application of ganoderma lucidum ketonediol in anti-colorectal cancer. Background technique [0002] Colorectal cancer includes colon cancer and rectal cancer. It refers to malignant tumors that occur in the cecum, ascending colon, transverse colon, descending colon, sigmoid colon, and rectum. It is one of the most common malignant tumors of the digestive tract. The latest data from the World Health Organization shows that the death toll of colorectal cancer ranks third in the number of cancer deaths. Every year, more than 1.2 million patients are diagnosed with colorectal cancer worldwide, and more than 700,000 patients die directly or indirectly from colorectal cancer. The incidence of colorectal cancer varies significantly in different regions, and the incidence of colorectal cancer is usually higher in men than in women. In addition, the incidence of colorectal can...

Claims

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Application Information

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IPC IPC(8): A61K31/575A61P35/00A61P35/04
CPCA61K31/575A61P35/00A61P35/04
Inventor 朱钦昌刘格贺震旦
Owner SHENZHEN UNIV
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