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Application of differentially expressed lncRNA to diagnosis and treatment of lung cancer

A lung cancer, expression level technology, applied in the field of biomedicine, can solve the problems of low early diagnosis rate, high late recurrence and metastasis rate, lack of sensitive and stable clinical drugs, etc., achieve high specificity and sensitivity, and high diagnostic efficiency

Inactive Publication Date: 2020-03-27
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, lung cancer still lacks effective and accurate early screening biomarkers and prevention targets, its early diagnosis rate is low but its late recurrence and metastasis rate is high, and it lacks sensitive and stable clinical drugs, so the overall survival rate of patients is low (CHEN W J, GAN T Q, QIN H, etal. Implication of downregulation and prospective pathway signaling of microRNA-375in lung squamous cell carcinoma[J]. Pathology, research and practice, 2017, 213(4):364-72.)

Method used

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  • Application of differentially expressed lncRNA to diagnosis and treatment of lung cancer
  • Application of differentially expressed lncRNA to diagnosis and treatment of lung cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 QPCR detection of the expression of AC105046.1 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 AC105046.1 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 AC105046.1. 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 pcDNA3.1(+)-1 was ligated for subsequent experiments.

[0082] 3. Transfection

[0083] The day before transfection, th...

Embodiment 3

[0090] Example 3 Effect of AC105046.1 on lung cancer cells

[0091] 1) MTT method to detect the effect of AC105046.1 on the proliferation of lung squamous cell carcinoma cells:

[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 AC105046.1 (0.376±0.043) was significantly lower than that of the blank control group...

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Abstract

The invention discloses application of differentially expressed lncRNA to diagnosis and treatment of lung cancer. Based on the down-regulation of expression of AC105046.1 in the lung cancer, the invention discloses application of a reagent for detecting AC105046.1 to preparation of a product for diagnosing the lung cancer and a corresponding product for diagnosing the lung cancer. On the basis that cell proliferation and migration can be influenced by regulating the expression level of AC105046.1, the invention discloses application of an AC105046.1 promoter to preparation of a pharmaceuticalcomposition for treating the lung cancer and a corresponding pharmaceutical composition.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of differentially expressed lncRNAs in the diagnosis and treatment of lung cancer. Background technique [0002] According to the data published in the global cancer statistics report in 2018 (SIEGEL RL, MILLER K D, JEMALA. Cancer statistics, 2018[J]. CA: a cancer journal for clinicians, 2018, 68(1):7-30.), lung cancer New cases ranked second among tumors, and death cases ranked first. At present, lung cancer still lacks effective and accurate biomarkers and prevention targets for early screening, its early diagnosis rate is low but its late recurrence and metastasis rate is high, and it lacks sensitive and stable clinical drugs, so the overall survival rate of patients is low (CHEN W J, GAN T Q, QIN H, etal. Implication of downregulation and prospective pathway signaling of microRNA-375in lung squamous cell carcinoma [J]. Pathology, research and practice, 2017, 213(4):36...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886A61K45/00A61K31/7105A61P35/00A61P11/00
CPCA61K31/7105A61K45/00A61P11/00A61P35/00C12Q1/6886C12Q2600/158C12Q2600/178
Inventor 杨承刚高舒欣魏琳
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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