Gastric cancer auxiliary diagnosis gene order and application thereof

A technology of auxiliary diagnosis and gene sequence, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve problems such as difficult to judge, and achieve the effect of reducing the toxic and side effects of treatment

Inactive Publication Date: 2008-02-20
SHANGHAI BIOCHIP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, through morphological typing, some situations that are difficult to judge may occur, and the current histopathological typing depends on the experience and knowledge of pathologists, and different pathologists may get different results

Method used

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  • Gastric cancer auxiliary diagnosis gene order and application thereof
  • Gastric cancer auxiliary diagnosis gene order and application thereof
  • Gastric cancer auxiliary diagnosis gene order and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The preparation of embodiment 1 clinical sample

[0032] 47 suspected cases of gastric cancer underwent endoscopic examination and biopsy, and pathological diagnosis was confirmed as 28 cases of gastric cancer and 19 cases of non-cancerous inflammation or other pathological diseases of the stomach. Uniform criteria for diagnosis. Lesions and contralateral non-lesional gastric epithelial tissues were collected and frozen in liquid nitrogen. This step was completed by the Ruijin Hospital Affiliated to Shanghai Jiaotong University and the Gastroenterology Department of Xinhua Hospital.

Embodiment 2

[0033] Extraction, quality detection and purification of embodiment 2 total RNA

[0034] (1) Extraction of total RNA

[0035] 47 RNA extractions were performed on the lesional gastric mucosa and distal non-lesional gastric mucosa of suspected cases of gastric cancer, using MRC's TRIzol kit, a conventional mammalian tissue RNA extraction method. Add 1ml TRIzol per 100mg of tissue, and fully break up the tissue pieces with an electric homogenizer. Add about 1 / 5 of the volume of chloroform, mix well by inverting up and down for about 1 minute, and let stand at room temperature for 5 minutes. After centrifugation at 12,000rpm for 15 minutes at 4°C, carefully remove the supernatant, transfer to a new 1.5ml centrifuge tube, add an equal volume of isopropanol, gently invert to mix, and let stand at room temperature for 5 minutes. After centrifugation at 12000 rpm at 4°C for 10 minutes, the supernatant was removed, 2 / 5 volume of 70% ethanol was added to the precipitate, and the prec...

Embodiment 3

[0040] Example 3 Gene expression profile experiment

[0041] (1) In vitro transcription

[0042] 47 Take 2 μg of total RNA extracted from each sample, and use the Agilent linear amp kit to amplify cRNA in vitro, that is, first configure 2 μg RNA, 5 μl T7 Promoter primer, and 11.5 μl of Rnase-free Water in a 1.5 ml centrifuge tube, 65 Denaturation at ℃ for 10 minutes, ice bath for 5 minutes. Then add 4μl 5×FirstStrand Buffer, 2μl 0.1M DTT, 1μl 10mM dNTP mix, 1μl MMLV RT, 0.5μl RNase OUT, keep at 40°C for 2h (hot cover 65°C), react to synthesize a large amount of cDNA first strand and second strand, and then On this basis, add 20 μl 4×Transcription Buffer, 6 μl 0.1M DTT, 16 μl NTP, 4 μl 25mM aa-UTP, 6.4 μl 50% PEG, 0.5 μl Rnase OUT, 0.6 μl Inorganic Pyrophosphatase (inorganic pyrophosphatase), 0.8 μl T7 RNA Polymerase ( Polymerase), 5.7 μl RNase-free Water and 80 μl system were incubated at 40°C for 2 hours to transcribe the cRNA chain. Purification was performed using a QIAG...

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Abstract

The invention discloses a group of gene sequence of gastric cancer auxiliary diagnosis and a gene chip and a kit aiming at the group of gene sequence. The invention also discloses the application of the group of gene sequence of gastric cancer auxiliary diagnosis. The gene sequence of gastric cancer auxiliary diagnosis applied in the invention can type gastric cancer from molecular level and helps clinical diagnosis, thus more effectively healing patients and reducing treatment side effect to the utmost extent.

Description

technical field [0001] The invention relates to the field of molecular diagnosis, in particular to a group of gene sequences for auxiliary diagnosis of gastric cancer and its application. Background technique [0002] One of the challenges in tumor treatment is how to classify tumors more accurately and choose an appropriate treatment plan on this basis. At present, tumor typing or classification is mainly based on morphological criteria, which has certain limitations. The clinical heterogeneity of tumors suggests that there may be different molecular subtypes, because tumors with the same morphology may have significant differences at the molecular level. Different gene expression characteristics, and thus different cellular and molecular biological behaviors, may produce different responses to the same treatment. As human biomedicine enters the era of molecular level, general pathological and morphological research can no longer fully meet the needs of tumor treatment. Th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11C12N15/12C40B40/06
Inventor 张庆华肖华胜韩峻松张兰晶渠翊杨燕青林利波
Owner SHANGHAI BIOCHIP
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