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Application of gastrin in inhibiting gastrointestinal tumor tumors

A tumor cell and gastrin technology, applied in the fields of pharmacology, cell biology, and molecular biology, can solve the problem of gastrin inhibiting tumor cells

Active Publication Date: 2010-11-24
SHANGHAI JIAOTONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the direct use of gastrin to inhibit tumor cells in clinical practice.

Method used

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  • Application of gastrin in inhibiting gastrointestinal tumor tumors
  • Application of gastrin in inhibiting gastrointestinal tumor tumors
  • Application of gastrin in inhibiting gastrointestinal tumor tumors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1. Gastrin inhibits the expression of gastric cancer cell lines AE1 and p16

[0024] 1. Treatment of gastric cancer cells SGC7901 with different concentrations of gastrin .

[0025] SGC7901 cells (gastric cancer cells involved in the present invention are all obtained from the Shanghai Institute of Biological Sciences, Chinese Academy of Sciences) in DMEM medium (Gibco BRL, Gaithersburg, ML) containing 10% fetal bovine serum (Gibco BRL, Gaithersburg, ML) containing 5% CO 2 -95%O 2 cultured in an incubator at 37°C. The SGC7901 cells were treated with 0.0001, 0.01, 0.1 and 1 μM gastrin for 72 hours and 0.1 μM gastrin for 6 hours, 8 hours, 12 hours and 24 hours.

[0026] 2. Detection of the expression of AE1 and p16: western-blot technology

[0027] 1) collecting cells;

[0028] 2) Wash twice with cold PBS, and discard as much residual PBS as possible;

[0029] 3) Resuspend the cells with an appropriate volume of pre-cooled normal lysis solution (SDS) (pr...

Embodiment 2

[0055] Example 2. Correlation between gastrin and expression of AE1 and p16

[0056] 1. Gastric cancer tissue samples (taken from Ruijin Hospital and Renji Hospital Affiliated to Shanghai Jiaotong University) : 4% paraformaldehyde fixed, paraffin embedded sections;

[0057] 2. Routine dewaxing of slices ;

[0058] 3. Detection of expression of gastrin, AE1 and p16 :Immunohistochemistry

[0059] 1) Wash with distilled water for 3min / 2 times;

[0060] 2) In order to reduce non-specific background staining caused by endogenous peroxidase, incubate the slices in Hydrogen Peroxide Block for 10-15 minutes;

[0061] 3) Wash with PBS buffer for 5min / 2 times;

[0062] 4) Drop primary antibody: anti-gastrin polyclonal antibody (MAIXIN-Bio, Fuzhou, China), anti-AE1 monoclonal antibody (Sigma, St.Louis, MO, USA) and anti-p16 polyclonal antibody (SantaCruz Biotechnology) , incubate the working solution at 37°C for 1-2 hours;

[0063] 5) Wash with PBS buffer for 5min / 2 times;

...

Embodiment 3

[0069] Example 3. MTT assay detects the inhibitory effect of gastrin on the viability of gastric cancer cells

[0070] 1. MTT experimental method

[0071] An equal amount of SGC7901 cells in the logarithmic phase was planted in two 60mm culture dishes (one dish for the control group and one dish for the experimental group), and the cell seeding density was about 10%. After 12 hours, the cells adhered to the wall, and the cells in the control group and the experimental group were changed once. Drugs were added (gastrin concentration: 0.1umol / L or 0.1nmol / L) when the experimental group changed the medium, which was marked as the 0th day. The cells of the control group and the experimental group were treated in the same way at the same time every day. On the fourth day, when the cell density of the control group and the experimental group reached about 80% (not piled up), it was subcultured, digested with 1ml 0.25% trypsin, and blown into a single cell suspension with 1.5ml cu...

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Abstract

The invention discloses a novel application of gastrin in inhibiting gastrointestinal tumor tumors. Gastric cancer target gene anion exchange protein 1 is interacted with tumor suppressor protein p16 so as to induce gastric cancer, however, 17 peptide gastrin (H-pGlu-Gly-Pro-Trp-Leu-Glu-Glu-Glu-Glu-Glu-Ala-Tyr-Gly-Trp-Met-Asp-Phe-NH2) can effectively inhibit expression of gastric cancer target gene AE1 and p16 and breaks through the traditional view that gastrin induces tumors, and clinical pathologies show that the gastrin has obvious negative correlation with AE1 and p16. The invention alsodiscloses that the gastrin with the concentration of 10-10mol / L-10-70mol / L has obvious inhibition effect on gastric cancer cell activity after a certain time, thus prompting that the gastrin has curing effect on gastric antrum cancer and providing a new direction for curing gastric cancer based on the test results.

Description

technical field [0001] The invention relates to the fields of molecular biology, cell biology and pharmacology, and more specifically, relates to a new application of gastrin. Background technique [0002] Gastric cancer ranks fourth among all malignant tumors in new cases each year, and ranks second among all causes of death due to tumors. There are approximately 1,000,000 gastric cancer cases and more than 850,000 deaths per year, and early diagnosis of gastric cancer Difficult, the 5-year survival rate is only 20%. The pathogenesis of gastric cancer has made progress in many aspects in recent years, including Helicobacter pylori infection, environmental factors, gene mutation and so on. Gastrin is a polypeptide hormone that is mainly secreted by G cells in the gastrointestinal tract and specifically binds to gastrin receptors. A large number of studies have shown that gastrin not only stimulates gastric acid secretion and has nutritional effects on gastrointestinal mucos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/22A61P35/00A61P1/00
Inventor 傅国辉
Owner SHANGHAI JIAOTONG UNIV SCHOOL OF MEDICINE
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