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Tissue chip and cellline chip for observation of cell signal conducting path

A signal transduction and cell line technology, applied in the field of tissue and cell chips for high-throughput detection and analysis of signal transduction pathways in different cells, can solve the problem of inability to directly detect changes in the expression of one or more genes in situ.

Inactive Publication Date: 2005-03-16
NEWORGEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing gene chips only immobilize DNA on the substrate to form a microarray, and cannot directly detect the expression changes of one or more genes in a certain cell in situ

Method used

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  • Tissue chip and cellline chip for observation of cell signal conducting path
  • Tissue chip and cellline chip for observation of cell signal conducting path
  • Tissue chip and cellline chip for observation of cell signal conducting path

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Phosphorylation status detection of Akt gene in normal liver tissue, liver cancer tissue, paracancerous liver tissue and liver cancer cell lines

[0073] 1. Materials and Methods

[0074] Cancer tissue-cell line chip: The tissue chip used in this study consists of 54 pairs of hepatocellular carcinoma and paracancerous tissues, normal liver tissues of 9 cases of accidental death, and subcutaneous tumor tissues of nude mice with 3 hepatoma cell lines Hep3B, SMMC7721, and BEL7402 Equal composition (see Table 1 and figure 1 ).

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Embodiment 2

[0110] Phosphorylation status detection of Smad1 gene in normal liver tissue, liver cancer tissue, paracancerous liver tissue and liver cancer cell lines

[0111] According to the method described in Example 1, immunohistochemical analysis was performed with antibodies against Smad1 phosphorylated protein and Smad1 total protein.

[0112] The results showed that the phosphorylation of Smad1 in the cytoplasm of cancer cells was weakly positive, the positive rate was 31.37% (16 / 51), and it was negative in the nucleus; while the cytoplasm of paracancerous liver and normal liver tissue were all positive in a moderate degree , the positive rate was 100% (57 / 57; 9 / 9); SMMC7721, BEL7402 and Hep3B showed moderate positive reaction in the cytoplasm.

[0113] The total protein of Smad1 in the cytoplasm of cancer cells was weakly positive, the positive rate was 31.37% (16 / 51), and the nucleus was also positive to varying degrees; while the weakly positive rate in the cytoplasm of adjacen...

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Abstract

The present invention provides tissue and cell chip for detecting and analyzing cell signal conducting path in various normal and disease tissues in high flux. The chip includes one substrate and one paraffin slice on one main surface of said substrate. The slice in 1-20 micron thick, and has 4-1000 sample applying holes, each for one kind of tissue or cell, and over 2 kinds of tissue and / or cell. The present invention provides the preparation process and use of the said chip, and may be used, especially, for detecting the difference in gene expression state and / or phosphorylation state of signal conducting path between different cells.

Description

technical field [0001] The invention belongs to the field of biotechnology. More specifically, it relates to a tissue and cell (line) chip for high-throughput detection and analysis of signal transduction pathways in different cells (such as various normal and disease tissue cells). Background technique [0002] Human genomics research is currently a hot spot in the world. With the progress of research, it is urgent to discover the changes of signal transduction pathways in various malignant tumor tissues and cells, and to determine new human genes and genes with known sequences, as well as the roles of various genes in various cellular pathways. [0003] In addition to large-scale sequencing of human chromosomal DNA and expressed sequence sequencing (EST) proteome methods, there is also a lack of a high-throughput method for detecting changes in cell signaling pathways. The existing gene chips only fix the DNA on the substrate to form a microarray, which cannot directly d...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/02
Inventor 顾健人李锦军
Owner NEWORGEN