Non-small cell lung cancer molecular marker and its application

A non-small cell lung cancer, isotope labeling technology, applied in the biological field, can solve the problems of low early diagnosis rate and unclear reasons for recurrence

Active Publication Date: 2011-11-16
CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

An important reason is that the early diagnosis rate is low, less than 2%, and more than 80% of patients are already in advanced stage when they see a doctor.
The cause of recurrence is also very unclear. Cur...

Method used

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  • Non-small cell lung cancer molecular marker and its application
  • Non-small cell lung cancer molecular marker and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1. Treatment of blood samples from patients with non-small cell lung cancer and normal control blood samples and preparation of peptides

[0049] The urea used in this example, 3-[(3-cholamidopropyl)-diethylammonium]-1-propanesulfonic acid (CHAPS), sodium dodecylsulfonate (SDS), dithiothreose Alcohol (DTT) and 4-hydroxyethylpiperazineethanesulfonic acid (HEPES) were purchased from Bio-Rad.

[0050] Blood from patients newly diagnosed with adenocarcinoma (lung adenocarcinoma, AD) or squamous cell carcinoma (squamous cellcarcinoma, SCC) (collected before treatment), and normal controls were collected. The specific information of the samples is shown in Table 1, Table 2 and Table 3 respectively. Among them, the 20 pairs of samples in Table 1 were paired according to factors such as gender, age, smoking status, and TNM stage. The MRM data of these samples were used for the prognosis analysis of lung squamous cell carcinoma. Table 2 shows the information of another...

Embodiment 2

[0065] Example 2, online integrated column chromatography fractionation, liquid chromatography tandem mass spectrometry analysis, protein identification and data analysis

[0066] The method of online integrated column chromatography fractionation and liquid chromatography tandem mass spectrometry is as follows: 11 pH gradients are used for fractionation on a strong cation exchange column (Column Technology Company). Then further fractionation on the reverse phase column (ColumnTechnology company), the mobile phase is 0.1% formic acid (v / v) solution (solution A) and 0.1% formic acid / acetonitrile (v / v) solution (solution B), within 115 minutes the solution B increased from 2% to 40%, detected on LTQ mass spectrometer (Thermo Finnigan).

[0067] The mass spectrometry data was searched by SEQUEST software in the International Protein Index (IPI) database (version number 3.22), and then used the Buildsummary software of our laboratory to search the positive and negative libraries ...

Embodiment 3

[0071] Example 3, Peptide Screening, Synthesis and Optimization of Detection Conditions for Transition and Quantitative Performance Analysis

[0072] According to the results of Example 2, combined with the screening criteria of peptides, the candidate peptides and Transition corresponding to the three proteins were determined. Then use the optimizer software of Agilent to optimize the experimental conditions of Transition, and at the same time select Transition with better signal intensity for protein quantification. The results are shown in Table 4. The corresponding coefficient of variation was calculated by performing 5 experimental repetitions on the mixed peptide samples, and the results are shown in Table 4.

[0073] Isotope-labeled Heavy Peptides 13 C-labeled leucine ([13C6]Leucine) and alanine ([13C3]Alanine), purchased from Sigma-Aldrich) and unlabeled light-labeled peptides were synthesized by Gil Biochemical (Shanghai) Co., Ltd., purity >95%.

[0074] Then dete...

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Abstract

The invention relates to a non-small cell lung cancer molecular marker and its application, specifically to an application of human leucine-rich alpha2 glycoprotein or its segment, human C4 binding protein or its segment, human serum amyloid protein or its segment, or the combination thereof in the preparation of a preparation for detecting whether an object has the non-small cell lung cancer or performing a prognosis treatment for those who suffer from the non-small cell lung cancer.

Description

technical field [0001] The invention belongs to the field of biotechnology. Specifically, the present invention relates to the three proteins of human leucine-rich α2 glycoprotein, human C4 binding protein and human serum amyloid protein used alone or in combination as molecular markers of non-small cell lung cancer and as non-small cell lung cancer Application of prognostic indicators. Background technique [0002] Lung cancer is one of the most common malignant tumors, and its incidence rate ranks first among all malignant tumors. In my country, lung cancer has become the first cause of death from malignant tumors, accounting for 22.7% of all malignant tumor deaths, and the morbidity and mortality are still rising. Between 2000 and 2005, the number of lung cancer patients in my country increased by 120,000. If effective control measures are not taken in time, it is estimated that the number of patients will reach 1 million by 2025, making China the world's largest lung c...

Claims

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Application Information

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IPC IPC(8): G01N33/68
Inventor 曾嵘刘延盛陈海泉罗晓阳徐孟杰
Owner CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI
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