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Application of diagnostic marker GATA4 in pancreas inflammatory cancer transformation

A diagnostic marker, pancreatitis technology, applied in the application field of the diagnostic marker GATA4 in the "inflammation and cancer transformation" of the pancreas, achieving the effect of excellent accuracy

Inactive Publication Date: 2021-04-09
SHANGHAI FIRST PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] There is no report about the application of the diagnostic marker GATA4 of the present invention in the "inflammation and cancer transformation" of the pancreas

Method used

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  • Application of diagnostic marker GATA4 in pancreas inflammatory cancer transformation
  • Application of diagnostic marker GATA4 in pancreas inflammatory cancer transformation
  • Application of diagnostic marker GATA4 in pancreas inflammatory cancer transformation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1 Pre-experiment

[0032] iTRAQ mass spectrometry screened differentially expressed genes in pancreatitis and pancreatic cancer tissue samples, suggesting that GATA4 was highly expressed in pancreatic cancer tissue.

[0033]Isotope-labeled relative and absolute quantification iTRAQ technology is a polypeptide in vitro labeling technology developed by Applied Biosystems Incorporation (ABI). This technology uses 4 or 8 isotope tags, by marking the specific amino acid sites of peptides (Peptide), and then performing tandem mass spectrometry analysis, it can flexibly compare the relative and absolute contents of proteins in various samples. In the iTRAQ experiment, the whole protein in the tissue cells was obtained through protein extraction, and the whole protein was enzymolyzed (Trypsin Digestion) into peptides under the action of trypsin, and the iTRAQ reagent labeled all the peptides obtained. iTRAQ reagent is mainly composed of 3 parts: reporter group (Repo...

Embodiment 2

[0034] Embodiment 2 clinical tissue sample experiment

[0035] This example studies the relationship between GATA4 and "inflammation-cancer transformation" in clinical tissue samples.

[0036] 1 method

[0037] 39 pancreatic cancer samples from 7 adjacent pancreatic cancer samples were taken from Shanghai First People's Hospital, and the expression of GATA4 in the clinical samples was analyzed: RT-PCR, Western-blot and immunohistochemical analysis were performed in clinical pancreatic cancer samples To further verify the expression of GATA4, the level of GATA4 and CD68 were detected by immunofluorescence;

[0038] 2 results

[0039] Using RT-PCR to analyze 7 pancreatic cancer samples and 39 pancreatic cancer samples, we found that the expression of GATA4 RNA was up-regulated compared with normal pancreatic tissue (P figure 2 ).

Embodiment 3

[0040] Example 3 Cell experiment

[0041] This example studies the relationship between GATA4 and "inflammation-cancer transformation" in cells.

[0042] (1) Correlation between the level of GATA4 and the level of CD68 (macrophage marker): Immunofluorescence detection in pancreatic cancer tissue found that the two can be co-expressed, suggesting that GATA4 may be involved in the regulation of inflammation in the development of pancreatic cancer;

[0043] (2) Promoting effect of LMCM on pancreatic cancer cells: Add different concentrations of lipopolysaccharide LPS (0, 50, 100 ng / ml) to the macrophage culture medium that induces THP-1 differentiation, observe and record the results.

[0044] (3) Expression of GATA4 in pancreatic cancer cells: RT-PCR and WB were used to detect the expression of GATA4 in various pancreatic cancer cell lines, and the results were observed and recorded.

[0045] (4) Interfering with GATA4 to observe the effects of LMCM with different LPS concentra...

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Abstract

The invention relates to application of a diagnostic marker GATA4 in pancreas inflammatory cancer transformation. According to the invention, immunofluorescence detection finds that the level of the GATA4 is positively correlated with the CD68 level, and prompts that the GATA4 may participate in inflammatory regulation in the pancreatic cancer development process. According to the experiment, lipopolysaccharide (LPS) with different concentrations is added into a macrophage culture medium LMCM for inducing THP-1 differentiation to treat pancreatic cancer cells; the result shows that up-regulation of the GATA4 level in the pancreatic cancer cells can be promoted, and proliferation, migration and invasion of the pancreatic cancer cells can be promoted; and meanwhile, overexpression of the GATA4 can obviously promote proliferation, migration and invasion ability of the pancreatic cancer cells induced by the LMCM with different LPS concentrations. This prompts that the GATA4 may be a key factor in the pancreatitis cancer transformation process. The invention provides a method for diagnosing pancreas inflammatory cancer transformation through molecular detection; the diagnosis speed is high; the cost is low; and the result is accurate and reliable.

Description

technical field [0001] The invention relates to the technical field of molecular diagnosis, specifically, the application of the diagnostic marker GATA4 in the "inflammation-cancer transformation" of the pancreas. Background technique [0002] Pancreatic ductal adenocarcinoma (PDAC) has a high degree of malignancy, difficult early diagnosis, and poor curative effect of existing treatment methods, so its mortality rate is high and its prognosis is extremely poor. Its five-year survival rate is only 5%. Although the introduction and application of new diagnostic and treatment methods have brought the morbidity and mortality of other cancers under control, the morbidity and mortality of pancreatic cancer continue to rise. In 2012, PDAC ranked first in the mortality rate of malignant tumors in the United States. 4, and by 2030 it is predicted to rise to the 2nd place. It is of great scientific significance and clinical value to study the initiating link of pancreatic cancer in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6883G01N33/574G01N33/68A61P35/04A61P35/00A61P1/18A61K45/00
CPCC12Q1/6886C12Q1/6883G01N33/57438G01N33/57484G01N33/6893A61K45/00A61P1/18A61P35/00A61P35/04C12Q2600/118C12Q2600/158G01N2800/067
Inventor 杨丽娟蒋魏亮陈聪颖黄丽沈杰
Owner SHANGHAI FIRST PEOPLES HOSPITAL
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