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Breast carcinoma transfer related gene chip establishment and its establishing method

A technology of gene chip and establishment method, applied in the field of gene chip preparation, to achieve good social and economic benefits

Inactive Publication Date: 2007-08-01
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is currently a lack of effective technical means for screening and identifying drugs that already have anti-breast cancer effects or whether newly developed drugs have the effect of inhibiting tumor metastasis

Method used

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  • Breast carcinoma transfer related gene chip establishment and its establishing method
  • Breast carcinoma transfer related gene chip establishment and its establishing method
  • Breast carcinoma transfer related gene chip establishment and its establishing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Selection of breast cancer metastasis-related genes in the gene chip of embodiment 1

[0023] Select the genes used in the gene chip of breast cancer metastasis-related genes: The selected genes include: ①Comparing the gene expression profiles of breast cancer MCF-7 cells and LM-MCF-7 cell lines by gene chip technology, the genes with differential expression Candidate genes were used to design gene chips; ②According to literature reports, genes related to breast cancer metastasis were selected. There are 218 genes in total, mainly related to oncogenes, growth factors and their receptors, metastasis accessory proteases, protease inhibitors, signal transduction molecules, metastasis suppressors, metastasis promoters, cell-cell and cell-matrix molecules , and internal reference genes such as ACTB, GAPD, LDHA and RPL9, etc.

Embodiment 2

[0024] The design of embodiment 2 gene chip probes

[0025] Design of gene chip probes: After determining the name of the target gene, search for the corresponding gene sequence on NCBI through the search number in GenBank, and find the coding region (ORF) of the gene. The hybridization region when designing the probe is located in the ORF. The basic principles of probe design: ①The nucleotide length of the probe is 40-60nt; ②Avoid the formation of hairpin structure inside the probe (continuous complementary sequence shall not be greater than 4nt); ③Avoid pairing between probes (continuous paired base ④Ensure that the Tm values ​​of the probes are basically similar; ⑤Apply the primer 5 primer design software to design the probes, and select the probes with higher scoring rates from the designed probes and send them back to the NCBI database for further analysis. BLAST alignment to ensure the specificity of the probes, so that they are only complementary to the gene without non...

Embodiment 3

[0026] Example 3 Preparation of gene chips related to breast cancer metastasis

[0027] Preparation steps of gene chip related to breast cancer metastasis:

[0028] ① According to the designed probe sequence, synthesize 218 60mer oligonucleotide (oligo) probes. When preparing the gene chip, positive control HEX, negative control 50% DMSO, external reference system Y1-Y8 and housekeeping genes (ACTB, GAPD, LDHA, RPL9) etc. were added respectively.

[0029] ②Dissolve the oligo probe in sterile water to a final concentration of 40 μM, and take 5 μl to the 384-well plate in which the sample is applied in a certain order.

[0030] ③ Apply the chip spotter to spot the synthesized probes on a sheet with an area of ​​75×25mm 2 and on amino-modified glass slides. The pitch of each gene point is 200 μm, and the diameter of the point is about 150 μm. The spotting is repeated 3 times. Each dot array has 26 rows and 27 columns, and each chip has four repeated gene dot arrays.

[0031] ④T...

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Abstract

The invention discloses the establishment of gene chip relative to breast cancer transition and the establishment method. The gene chip comprises 218 genes, in which includes breast cancer transmission relative gene got by comparaing human breast cancer MCF-7 with human breast cancer system LM-MCF-7 (be awarded with patent for invention in China, ZL 03130264.5) with high transmission tendency, and breast cancer transmission relative gene reported by papers. Synthesizes oligonucleotide (oligo) with 218 genes, and spots molecular probe on plate modified by amino. Said prepared breast cancer transmission relative gene chip is mainly used for sifting anti- breast cancer transmission medicine, and states the molecular principle of medicine action, and can aslo be used for early- phase diagnosis for breast cancer transmission, assisting individual treatment program and prognosis judgement.

Description

technical field [0001] The invention relates to the design of gene chip probes for breast cancer metastasis-related genes, the preparation of gene chips, and the like. The gene chip is mainly used for the screening of anti-breast cancer metastasis drugs, early diagnosis of breast cancer metastasis, auxiliary individualized treatment plan and prognosis judgment, etc., belonging to the field of biotechnology. Background technique [0002] The occurrence and development of breast cancer is a complex process involving multiple factors, multiple stages and multiple genes, and metastasis is the main cause of death of patients. If drugs against breast cancer metastasis can be obtained, it means that breast cancer metastasis can be treated more effectively. However, currently there is a lack of effective technical means for screening and identifying drugs that already have anti-breast cancer effects or whether newly developed drugs have the effect of inhibiting tumor metastasis. T...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 叶丽虹张晓东王长晔
Owner NANKAI UNIV
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