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A diagnostic marker for early metastasis of lung cancer and its kit and application

A diagnostic kit and technology for lung cancer, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve problems such as false positives, lncRNA clinical significance, function and mechanism not clarified, and achieve specificity Strong, fast and convenient detection, high reliability

Active Publication Date: 2021-09-21
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, spiral CT is the most commonly used method to detect lung cancer clinically, but there is a high rate of false positives; more than 50% of lung cancer patients are diagnosed at an advanced stage, and the median survival time of advanced lung cancer patients is less than 18 months
In addition, tumor-specific lncRNAs can be found in the detection of precancerous lesions and lesion tissues, blood, urine, and saliva of tumor patients, suggesting that they can be used as a potential tumor diagnosis and prognostic factor. The specific clinical significance, function, and mechanism of development have not yet been elucidated

Method used

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  • A diagnostic marker for early metastasis of lung cancer and its kit and application
  • A diagnostic marker for early metastasis of lung cancer and its kit and application
  • A diagnostic marker for early metastasis of lung cancer and its kit and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1 Verification of LncRNA ENSG00000270607, a characteristic lncRNA of lung cancer metastasis

[0057] The team of the present inventors found through a lot of previous research that LncRNAENSG00000270607 shown in SEQ ID NO: 1 is related to the diagnosis of lung cancer metastasis and can be used as a blood molecular diagnostic marker for early metastasis of lung cancer. The serum samples from 154 outpatients or inpatients with non-small cell lung cancer in Sun Yat-Sen University Cancer Hospital from June 2017 to September 2017 were used to detect LncRNA ENSG00000270607 by probe method PCR, so as to verify the characteristic LncRNA ENSG00000270607 of lung cancer metastasis . Specifically include the following steps:

[0058] 1. Extraction of total RNA from plasma samples (TRIZOL LS method) and determination of RNA concentration

[0059] (1) Sample homogenization: take 0.25mL sample and add 0.75ml TRIzol LS reagent, blow up and down with a pipette several times to...

Embodiment 2

[0104] Example 2 In vitro invasion test of small chamber to detect the effect of LncRNA ENSG00000270607 on the invasion ability of non-small cell lung cancer cells in vitro

[0105] (1) Pretreatment of samples

[0106] First, take out the Matrigel (BD Company) stored at -80°C, put it in an ice box and embed it with ice, and then put the ice box in a chromatographic cabinet at 4°C overnight to thaw the frozen Matrigel; take the thawed Matrigel with the basal medium (i.e. medium without fetal bovine serum) at V Matrigel / V 基础培养基 =1 / 9 The ratio was fully diluted and the mixture was filled into EP tubes with a volume of 1.5 mL and placed on ice.

[0107] (2) Small chamber in vitro invasion test detection

[0108] Place a small chamber (24Well Transwell Permeable Supports) in a 24-well plate, take 50 μL of the above-mentioned diluted mixture, spread it evenly on the bottom of the upper layer of the small chamber, and pay attention not to generate air bubbles; keep it in an incub...

Embodiment 3

[0111] Example 3 Diagnostic value of LncRNA ENSG00000270607 in patients with lung cancer metastasis

[0112] A diagnostic kit for early metastasis of lung cancer, including specific amplification primer pairs and probes for detecting LncRNA ENSG00000270607 markers, and primer pairs and probes for amplifying internal reference genes, including upstream primer F and downstream primer R and probes. The amplification primer pair and probe of the LncRNA ENSG00000270607 and the primer pair and probe of the internal reference gene are as follows respectively:

[0113] LNCRNA ENSG00000270607:

[0114] LNCRNA ENSG00000270607-F (SEQ ID NO: 2):

[0115] 5'-TGGGCAAGTTATTTAAGC-3';

[0116] LNCRNA ENSG00000270607-R (SEQ ID NO: 3):

[0117] 5'-CAACCATCTCATTTAATCCA-3';

[0118] LNCRNA ENSG00000270607 fluorescent probe (SEQ ID NO: 4):

[0119] 5'-FAM-AATGTCGTCAAAGCCTCTCACCT-BHQ1-3'.

[0120] Internal reference ACTB:

[0121] ACTB-F (SEQ ID NO:5): 5'-TCACCCACACTGTGCCCATCTACGA-3';

[01...

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Abstract

The invention belongs to the technical field of disease diagnosis and prevention, and specifically discloses an LncRNA ENSG00000270607 marker for early metastasis diagnosis of lung cancer, a kit and application thereof. The sequence of the LncRNA ENSG00000270607 is shown in SEQ ID NO: 1, and it can be used as a molecular marker to distinguish whether lung cancer patients transfer. The kit provided by the invention is fast and convenient, has high accuracy and sensitivity, and has strong specificity. The diagnosis and prediction of early metastasis of lung cancer can be widely used in the diagnosis and treatment of early metastasis of lung cancer in the medical industry.

Description

technical field [0001] The present invention relates to the technical field of disease diagnosis and prevention, and more specifically, relates to a lung cancer early metastasis diagnostic marker LncRNA ENSG00000270607 and its kit and application. Background technique [0002] Lung cancer is a malignant tumor with the highest morbidity and mortality worldwide. According to the "World Cancer Report" recently released by the World Health Organization (WHO), there were 1.8 million new cases of lung cancer in the world in 2012, accounting for 13% of the incidence of cancer, and lung cancer-related deaths accounted for nearly all cancer-related deaths. one third. In the past 30 years, the mortality rate of lung cancer in my country has increased by 46.5%, and the incidence rate has increased by an average of 26.9% per year. On average, nearly 400,000 patient deaths per year are related to lung cancer. At present, the clinical treatment effect of non-small cell lung cancer is st...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 黎孟枫管洪宇吴姗姗刘蕾向涛朱勋吴珏珩刘湜桦朱婷刘帮洞于暕辰张乐
Owner SUN YAT SEN UNIV
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