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Application of leukotriene A4 hydrolase as lung cancer early marker

A hydrolytic enzyme and leukotriene technology, applied in the field of laboratory medicine, can solve the problems of no precancerous lesions, insufficient sensitivity, and no improvement in confocal microscopy, and achieve high sensitivity and high specificity

Pending Publication Date: 2021-10-26
YANSHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Recent developments in imaging-based screening techniques such as spiral computed tomography (CT), optical coherence tomography, positron emission tomography (PET), virtual bronchoscopy, autofluorescence bronchoscopy, and confocal microscopy have also not been shown to improve this signs of this condition, possibly because the morphological criteria used in imaging methods were not sensitive enough in the precancerous stages of lung cancer development and it had no morphologically identifiable precancerous lesions

Method used

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  • Application of leukotriene A4 hydrolase as lung cancer early marker
  • Application of leukotriene A4 hydrolase as lung cancer early marker
  • Application of leukotriene A4 hydrolase as lung cancer early marker

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Experimental program
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Effect test

Embodiment 1

[0027] Using the SELEX (exponential enrichment systematic evolution of ligands) Screening of aptamers obtained from the serum of lung cancer, the nucleic acid sequence of the aptamer as follows:

[0028] 5'-CTATAGCAATGGTACGGTACTTCCTAGTTGAGCATTATACACATTGA CTG GGCTGTTCTTCTGTCAAAAGTGCACGCTACTTTGCTAA-3 '(SEQ ID NO.2), a nucleic acid aptamer quantum dot labeled aptamer prepared per tube 1mL, 1mL was added taken out of each 500μL ethanol (stock solution: ethanol = 1: 2 ), 12000g, 4 ℃ centrifuged 10min, the supernatant was discarded, 1mL of ultrapure water was added to dissolve, i.e. 5nM. 95 deg.] C prior to use denaturation 5min, the ice bath was immediately 5min, so ssDNA restored three-dimensional structure.

[0029] Serum collected from the First Hospital of Qinhuangdao, in which 10 patients with lung cancer, 10 patients with pneumonia and 10 healthy people, 10 cases of colorectal cancer. Serum four groups were treated as follows:

[0030] After the mixed serum, 15000g, 4 ℃ centrifug...

Embodiment 2

[0035] Pull-Down experiments using tumor markers identified

[0036] Lung aptamers with streptavidin-coated with the biotinylated magnetic beads are incubated 37 ℃ 1h, eluting formamide, DEPC water degeneration, PBS + Mg 2+ Secondary degeneration. The aptamer bound magnetic beads with biotinylated rapid processing are well broth A549 cells and lung microvascular endothelial cells (HMVECs) the culture was incubated for 1h at 37 ℃, eluted with PBS, eluted under silver staining and protein sequencing, using air as a blank beads. Silver staining results figure 2 , The sequencing results image 3 , The sequencing of the resulting protein identification leukotriene A4 hydrolase enzyme protein sequence as shown in SEQ ID NO.1. Depend on figure 2 As can be seen, the present invention is obtained by screening a nucleic acid aptamer capable of specifically fished lung cancer proteins.

Embodiment 3

[0038] March 2019 to June 2021 were collected from the First Hospital of Qinhuangdao City Hospital, a total of 100 cases of peripheral blood of volunteers, 20 patients with lung cancer, 20 patients with pneumonia, 20 cases of healthy people, 20 patients with colorectal cancer and cervical 20 cases of cancer. No significant difference among the five groups mean age between groups of volunteers and gender, can be used for comparison between groups. Volunteers are obtaining informed consent, in which patients with lung cancer, colorectal cancer patients and cancer patients were confirmed by pathological findings organization, patients with pneumonia were confirmed by CT.

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Abstract

The invention provides application of leukotriene A4 hydrolase as a lung cancer early marker, which belongs to the technical field of laboratory medicine. The invention provides application of leukotriene A4 hydrolase as a marker in preparation of a product for evaluating lung cancer disease risk. What is found is that the expression quantity of leukotriene A4 hydrolase in lung cancer serum is higher than that of normal serum, and leukotriene A4 hydrolase can serve as a marker for lung cancer diagnosis and is used for auxiliary diagnosis and targeted therapy of lung cancer or fundamental research related to occurrence, development and progress of lung cancer diseases. The method is simple, rapid and sensitive, is suitable for auxiliary diagnosis of preclinical lung cancer and tumor bio-oriented treatment, and has wide clinical application prospects and basic application prospects.

Description

Technical field [0001] The present invention belongs to the technical field of medical testing, in particular, it relates to leukotriene A4 hydrolase enzymes applied as an early marker of lung cancer. Background technique [0002] 5-year survival rate of lung cancer is less than 15%, the main reason for the high mortality rate of lung cancer is most lung cancer patients are diagnosed in advanced treatment success rate is low. In all subtypes of lung cancer, small cell lung cancer (small cell lungcancer, SCLC) as a distinct clinical entity, its highest tendency to spread early, the shortest median survival time (7-12 months). Lung cancer patient survival depends on early detection and effective treatment, although the sputum cytology prior to use and a chest X-ray is detected early clinical trials may be used for early diagnosis of lung cancer, overall mortality but did not decline. Recently based screening techniques for imaging such as spiral computed tomography (CT), optical co...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/573
CPCG01N33/57423G01N33/573G01N33/57488G01N2333/914
Inventor 栗坤郭圆斌石明刘志伟李健
Owner YANSHAN UNIV
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