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Biomarker and application thereof in lung cancer treatment

A lung cancer and drug technology, applied in the field of biomedicine, to achieve the effect of high diagnostic efficiency, high specificity and sensitivity

Inactive Publication Date: 2020-04-21
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although the conventional methods widely used in clinical practice have certain sensitivity and accuracy for the detection of early malignant tumors, the detection of lncRNA expression levels can complement these conventional methods, and it has shown that it may become a new type of high-sensitivity biological tumor. The potential of biomarkers

Method used

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  • Biomarker and application thereof in lung cancer treatment
  • Biomarker and application thereof in lung cancer treatment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 QPCR detection of the expression of AC079630.1 in lung cancer

[0054] 1. Collect samples

[0055] 46 pairs of lung squamous cell carcinoma tissues and their corresponding paracancerous tissue samples were collected.

[0056] Inclusion criteria:

[0057] 1) The pathological diagnosis is lung squamous cell carcinoma, 2) The RNA expression level and clinicopathological information of the sample are complete.

[0058] Exclusion criteria:

[0059] 1) Suffering from other malignant tumors other than squamous cell carcinoma of the lung, 2) The case has received radiotherapy, chemotherapy and targeted drug therapy before specimen collection.

[0060] 2. RNA sample preparation and quantitative analysis

[0061] Use Trizol reagent to extract RNA from sample tissue, the steps are as follows:

[0062] Add 1mL Trizol to a glass homogenate bottle in an ultra-clean bench, weigh 50-100mg of tissue into the glass homogenate bottle, adjust the rotation speed to about 1500...

Embodiment 2

[0077] Example 2 Overexpression of AC079630.1 gene

[0078] 1. Cell culture

[0079] The lung squamous cell line H2170 cells were cultured in DMEM medium supplemented with 10% fetal bovine serum and P / S. For adherent growth of cells, place in conditions of 5% CO 2 , Cultivated in a constant temperature incubator with a humidity of 37°C.

[0080] 2. Construction of gene overexpression vector

[0081] Specific PCR amplification primers were synthesized according to the sequence of AC079630.1, and two restriction enzyme sites, KpnI and XhoI, were added to the 5' end primer and the 3' end primer, respectively. The cDNA extracted and reverse-transcribed from a lung squamous cell carcinoma patient was used as an amplification template, and the above cDNA sequence was double-digested with restriction endonucleases KpnI and XhoI and then inserted into the eukaryotic cell expression vector pcDNA3 that was double-digested with KpnI and XhoI .1(+), the obtained recombinant vector pcD...

Embodiment 3

[0090] Example 3 Effect of AC079630.1 on lung cancer cells

[0091] 1) The effect of AC079630.1 on the proliferation of lung squamous cell carcinoma cells was detected by MTT method:

[0092] Wash the transfected cell lines with PBS, remove dead cells, digest with trypsin, prepare a single cell suspension, dilute the cell suspension, and adjust the density to 5×10 4 cells / ml, and then inoculate in 96-well culture plate, add 100 μl of cell suspension to each well, put the 96-well culture plate in 37°C, 5% CO 2 Continue culturing in a cell culture box for 48 hours, add 1×MTT to each well, continue culturing for 4 hours, discard the supernatant, add DMSO to each inner well, shake for 10 minutes, and measure each well at a wavelength of 450nm with a microplate reader. OD value of the well.

[0093] MTT results showed that the cell growth and proliferation rate of the experimental group overexpressing AC079630.1 (0.321±0.033) was significantly lower than that of the blank control...

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Abstract

The invention relates to a biomarker and an application thereof in lung cancer treatment. According to the invention, the lung cancer can be judged by detecting the expression level of AC079630.1, theAUC value is 0.926, high diagnosis efficiency is achieved, the cell experiment overexpression of AC079630.1 remarkably inhibits the proliferation activity and migration capacity of cells, and the AC079630.1 can be used as a target to be applied to lung cancer treatment.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a biomarker and its application in lung cancer. Background technique [0002] Lung cancer is a public health issue that people are very concerned about in today's world. According to Cancer statistics, the cancer data released in 2018 (SIEGEL RL, MILLER K D, JEMAL A. Cancer statistics, 2018[J].CA: a cancer journal for clinicians, 2018 ,68(1):7-30), the estimated new cases of lung cancer in 2018 ranked second among tumors, and the estimated death cases ranked first, which is a major public health problem. The majority of lung cancers are non-small cell lung cancer (NSCLC), accounting for about 85% of lung cancers, which contains two main histological subtypes: lung squamous cell carcinoma (LUSC, accounting for 25-30%) and lung adenocarcinoma. Lung adenocarcinoma, LUAD, accounting for 40% At present, lung cancer still lacks effective and accurate biomarkers for early screening and prevent...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/158
Inventor 杨承刚高舒欣魏琳
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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