Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Use of neuritis auxin in preparing antineoplastic medicine

An anti-tumor drug and neurite technology, applied in the field of genetic engineering and medicine, can solve the problems of unstable physiological habits of cells, complex and changeable genetic background of tumor cells, etc.

Inactive Publication Date: 2008-11-19
FUDAN UNIV
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although both are rapidly proliferating cell populations, the genetic background of tumor cells is complex and changeable, the gene expression profiles vary widely, and the physiological habits of cells are quite unstable, while endothelial cells are relatively stable in all aspects

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Use of neuritis auxin in preparing antineoplastic medicine
  • Use of neuritis auxin in preparing antineoplastic medicine
  • Use of neuritis auxin in preparing antineoplastic medicine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1. Cell supernatant collection

[0059] 1) Take the cells cultured to the logarithmic growth phase, trypsinize, collect in a centrifuge tube and count, and centrifuge. Plant the required number of cells in a 60mm petri dish.

[0060] 2) After 24 hours, change to serum-free medium

[0061] 3) After 24, collect the serum-free medium, centrifuge at 3,000 rpm for 10 minutes to collect the supernatant

[0062] 4) Take 200 microliters of cell culture supernatant and add 2X loading buffer, denature at 98°C for 5 minutes. After denaturation, the sample and the cell sample are loaded together for electrophoresis detection.

[0063] The results showed that a variety of tumor cell lines can secrete neurites, including Huh-7, QGY, SK-Hep1, HepG 2 The secretion of cell lines is relatively high, while that of Focus, YY and other cell lines is relatively low.

Embodiment 2

[0064] Example 2. Expression of NRN1 full-length protein in Pichia pastoris

[0065] 2.1 Conversion

[0066] 1) Extract a large amount of PIC9K plasmid containing NRN-HIS, digest the plasmid with Sac I single enzyme, recover and purify.

[0067] 2) Inoculate GS115 in YPD medium, shake culture in 30 degree incubator to OD600 value 1.3-1.5.

[0068] 3) Centrifuge at 1,500 rpm for 5 minutes, discard the supernatant, add 30 ml of ice-cold water to resuspend,

[0069] 4) Centrifuge at 1,500 rpm for 5 min, discard the supernatant, and add 10 ml ice-cold water to resuspend

[0070] 5) Centrifuge at 1,500 rpm for 5 minutes, discard the supernatant, and add 2.5 ml of ice-cold sorbitol to resuspend

[0071] 6) Centrifuge at 1,500 rpm for 5 minutes, discard the supernatant, add 0.5 ml ice-cold sorbitol to resuspend, and the OD600 value is 50-60

[0072] 7) Take 80 microliters of yeast competent cells and add 5-20 micrograms of linearized DNA (dissolved in 5-10 microliters of TE)

[0073] 8) ...

Embodiment 3

[0110] Example 3. Cell Proliferation Experiment

[0111] The cell proliferation was measured by MTS method.

[0112] After the cells were digested and resuspended, they were planted in 96-wells (3×103 cells / well), and left to stand for 5 hours.

[0113] After the cells adhere to the wall, remove the medium, wash twice with PBS, and then add serum-free medium to culture for 24 hours.

[0114] Add neurite protein solution (the protein is diluted with DMEM serum-free) with final concentrations of 1 nM, 10 nM, and 100 nM to the culture wells, and the final culture medium volume per well is 100 μl. Only DMEM serum-free culture wells were used as blank control, wells containing PBS and filtrate were used as negative controls, and 1% FBS was used as positive control.

[0115] After culturing for 48 hours, the medium was removed, and 100 μl DMEM serum-free medium was added.

[0116] Add 20μl of MTS / PMS mixture to each well and continue to incubate for 3 hours to develop color.

[0117] Sh...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention belongs to the field of a biological medicine. The invention provides the new use of human protein neurite growth substance, namely, the application of the human protein neurite growth substance in preparing antineoplastic drugs. The human protein neurite growth substance shows obvious up-regulated expression in such malignant tumors as lung cancer, liver cancer, breast cancer, alimentary tract adenocarcinoma and so on, and can promote the tumor formation of mice. Accordingly, reducing the expression of neurite growth substance means inhibiting the growth of tumor. The human protein neurite growth substance provided by the invention can be used for early diagnosis of tumors, and can be used as a medicine for medicinal target screening and inhibiting the growth of tumor. The monoclonal antibody of the protein neurite growth substance can be used for preventing tumor blood vessels from growing, thus inhibiting the development of tumors. The invention further provides a kit and a biochip used for screening antineoplastic drugs.

Description

Technical field [0001] The present invention relates to the fields of genetic engineering and medicine. Specifically, the present invention relates to the application of human protein neurite outgrowth in the preparation of anti-tumor drugs. Background technique [0002] Neuritin (NRN1) is a neurotrophic factor discovered in 1997 with a full length of 142aa and a molecular weight of 15.3KD. It is mainly expressed in satellite cells of the brain, liver, and skeletal muscle, and a small amount is also expressed in the heart and lung. Phosphoinositide anchoring point (GPI), anchored on the cell membrane, can be separated from the membrane by the action of phospholipase C (PLC) and distributed in body fluids in a soluble form. Acts as a cytokine. Neurite growth factor has a strong role in promoting neurite growth, synaptic connection and maturation. The recombinantly expressed neurite growth factor protein stimulates primary cultured hippocampal neurons and cerebral cortex cells, whi...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): A61K38/18G01N33/68G01N33/15A61P35/00
Inventor 余龙秦波仙玲玲韩丁丁杨磊
Owner FUDAN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com