Screening method of lung cancer specific transcription regulation and control element

A transcriptional regulatory element and screening method technology, applied in the field of molecular biology, can solve the problems of low resolution and difficult to determine the core region of the screening sequence, etc., and achieve the effects of enhanced expression, simple operation, and high expression
CN105506062AInactive Publication Date: 2016-04-20HUAQIAO UNIVERSITY
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
HUAQIAO UNIVERSITY
Publication Date
2016-04-20
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention provides a screening method of a lung cancer specific transcription regulation and control element. The method comprises the following steps: acquiring epigenetic information in an area about 27kb long surrounding a DKK1 gene locus in a non-small cell lung cancer A549 cell by virtue of a UCSC database, predicting and obtaining 12 candidate enhancer transcription regulation and control element fragments, and respectively inserting the 12 enhancer transcription regulation and control element fragments to the upstream of a near-end promoter of DKK1 to successfully establish luciferase recombinant plasmids; and respectively transfecting the luciferase recombinant plasmids to the non-small cell lung cancer A549 cell, a non-small cell lung cancer H460 cell, a small cell lung cancer H446 cell, an esophagus cancer Eca-109 cell, a normal liver cell Chang liver and a human embryonic kidney cell HEK293, detecting the activity of relevant candidate elements by virtue of a dual-luciferase reporter gene detection system, and screening the regulation and control element having relatively high enhancing expression function on the DKK1 promoter. The screening method has the characteristics of simple operation.
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Description

【Technical field】

[0001] The invention belongs to the field of molecular biology, relates to gene expression regulation, and specifically relates to a screening method for lung cancer-specific transcription regulation elements. 【Background technique】

[0002] DKK1 is an antagonist of the canonical wnt signaling pathway, it can bind to Lrp5 / 6 receptor protein, and produce competitive inhibitory effect on the wnt signaling pathway. In the detection of the serum concentration of DKK1, compared with other malignant tumors, benign lung diseases and healthy controls, the serum DKK1 concentration of lung cancer patients is significantly higher, which can be used as a serum marker for the diagnosis of lung cancer, but the study found that the DKK1 promoter is in The activity in lung cancer was not very high. The space- and time-specific expression regulation of eukaryotic genes is an extremely complicated process. Some DNA sequences involved in the regulation of gene transcription ...

Claims

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