MicroRNA biological markers for early lung cancer diagnosis and application thereof

A biomarker, early diagnosis technology, applied in recombinant DNA technology, DNA/RNA fragment, microbial determination/examination, etc., can solve the problems of low sensitivity and specificity, patient exposure, etc., and achieve the effect of high diagnostic value

Active Publication Date: 2014-01-01
SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, CT scans may expose patients to harmful radiation, leading to more tumors
Several protein markers, such as cytokeratin 19 fragment (CYFRA21-1), carcinoembryonic antigen (CEA), neuron-specific enolase (NSE), etc., are used for non-surgical diagnosis of lung cancer, but the low sensitivity and specificity

Method used

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  • MicroRNA biological markers for early lung cancer diagnosis and application thereof
  • MicroRNA biological markers for early lung cancer diagnosis and application thereof
  • MicroRNA biological markers for early lung cancer diagnosis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Initial selection of markers in a small number of serum samples

[0048] We first detected 16 miRNAs (has-miR-21, has-miR-155, has-miR-205, has-miR-20a, hsa- miR-200b, hsa-miR-375, hsa-miR-486-5p, hsa-miR-186, hsa-miR-186-3p, hsa-miR-34c, hsa-miR-34c-3p, hsa-miR- 126, hsa-miR-34a, hsa-miR-125a-5p, hsa-miR-193a-5p and hsa-miR-25) expression. The results showed that hsa-miR-125a-5p, hsa-miR-193a-5p, hsa-miR-25 and hsa-miR-126 were down-regulated and hsa-miR-34a was up-regulated in lung cancer serum compared with normal controls expression (p36) to be accurately quantified. has-miR-21, has-miR-155, has-miR-205, has-miR-20a, hsa-miR-375, hsa-miR-486-5p, hsa-miR-186-3p and hsa-miR- There was no significant difference in the expression of 186 in serum samples of early stage lung cancer and normal serum samples (p>0.05) (Figure 2).

Embodiment 2

[0050] Further validation of miRNA markers

[0051] In order to verify the diagnostic value of the five miRNAs (hsa-miR-125a-5p, hsa-miR-193a-5p, hsa-miR-25, hsa-miR-126 and hsa-miR-34a) screened out initially, 106 The expression levels of these five miRNAs were further verified in the serum samples of 2 cases. The 48 samples from the primary screening step were also included in the statistical analysis of the validation step (the 106 serum samples here included 52 serum samples from stage I / II lung cancer patients and 54 serum samples from normal controls).

[0052] The principle of quantitative PCR detection of miRNA by probe method is as follows: figure 1 As shown, the primers and probes used in the quantitative PCR of each miRNA are as follows:

[0053] Table 1. miRNA sequences, reverse transcription primers, quantitative PCR pre-primers, universal post-primers and probe sequences

[0054]

[0055]

[0056] Note: The underlined sequence is the same sequence as the...

Embodiment 3

[0061] Receiver operating characteristic (ROC) curve analysis

[0062]A ROC curve was constructed to compare the diagnostic ability of five serum miRNAs in distinguishing early lung cancer patients from healthy controls. The area under the curve (AUC) of the five miRNAs ROC is as follows: hsa-miR-125a-5p, 0.840 (95% confidence interval, 0.760-0.920); hsa-miR-193a-5p, 0.819 (95% confidence interval, 0.736 -0.902); hsa-miR-25, 0.814 (95% confidence interval, 0.734-0.894); hsa-miR-126, 0.843 (95% confidence interval, 0.765-0.920); hsa-miR-34a, 0.738 (95% Confidence interval, 0.644-0.832) (Fig. 4A-4E). Under the optimal cutoff value, the sensitivity and specificity of miRNAs were as follows: hsa-miR-125a-5p, 83.3% and 82.7%, respectively; hsa-miR-193a, 64.8% and 94.2%; hsa-miR-25, 90.7% and 61.5%; hsa-miR-126, 70.4% and 82.5%; hsa-miR-34a, 67.3% and 74.1%. The combined AUC of these five miRNAs could reach 0.966, and the sensitivity and specificity were 92.3% and 90.7%, respecti...

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Abstract

The invention relates to the field of biological detection, particularly to a group of microRNA biological markers for early lung cancer diagnosis and an application thereof. The microRNA biological marker comprises hsa-miR-125a-5p, hsa-miR-193a-5p, hsa-miR-25, hsa-miR-126 and hsa-miR-34a. The serum miRNA biological markers comprising the 5 miRNAs can not only be particularly applied to early lung cancer detection, but also be applied to advanced lung cancer detection.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a group of microRNA biomarkers for early diagnosis of lung cancer and applications thereof. Background technique [0002] Lung cancer is the leading cause of cancer mortality worldwide, and its 5-year survival rate has improved only marginally over the past 3 years. Before reaching the advanced stage, lung cancer has no obvious clinical symptoms. More than 75% of lung cancer patients are diagnosed with locally advanced or metastatic lung cancer, and the late diagnosis of three quarters of lung cancer patients is the main reason for the low survival rate. The 5-year survival rate of patients with early-stage non-small cell lung cancer (NSCLC) is approximately 50%-70%, compared with 5%-15% and less than 2% for patients with stage III and stage IV NSCLC, respectively. Early diagnosis of lung cancer patients can effectively reduce their mortality. Chest x-rays and sputum cytolo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 毛红菊王萍张宏莲杨大伟洪群英金庆辉白春学赵建龙
Owner SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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