Marker for diagnosing and treating lung adenocarcinoma

A lung adenocarcinoma and drug technology, applied in the field of biomedicine, can solve the problems of unclear molecular biological mechanism of lung adenocarcinoma, difficult to distinguish, poor prognosis of lung adenocarcinoma, etc.

Active Publication Date: 2016-06-01
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing tests are difficult to distinguish lung adenocarcinoma from other types of lung tumors and pleural mesothelioma
[0004] Lung adenocarcinoma is poorly sensitive to radiotherapy and chemotherapy, and most patients are already in the advanced stage of the disease when they see a doctor, and surgical treatment is limited, so the prognosis of lung adenocarcinoma is poor
Although a large number of previous studies have attempted to describe and explain the specific causes of lung adenocarcinoma, the molecular biological mechanism of lung adenocarcinoma is still unclear.

Method used

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  • Marker for diagnosing and treating lung adenocarcinoma
  • Marker for diagnosing and treating lung adenocarcinoma
  • Marker for diagnosing and treating lung adenocarcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1 Screening for Gene Markers Related to Lung Adenocarcinoma

[0062] 1. Sample collection

[0063] Paracancerous tissue and lung adenocarcinoma tissue samples were collected from 8 cases of lung adenocarcinoma. The above-mentioned samples are surgical resection specimens of patients with lung adenocarcinoma, and all the above-mentioned samples were obtained with the consent of the organizational ethics committee.

[0064] 2. RNA sample preparation (using miRNAkit for operation)

[0065] The tissues obtained above were shredded, put into liquid nitrogen and ground into powder, and RNA was extracted and isolated according to the instructions in the kit. details as follows:

[0066] 1) Isolation of RNA:

[0067] A. Add in tissue homogenate or cells Reagent II 1ml;

[0068] B. Stand at room temperature for 3 minutes, add 0.2ml chloroform and shake vigorously for 15 seconds;

[0069] C. Place on ice for 10 minutes;

[0070] D. Centrifuge at 12000g for 15mi...

Embodiment 2

[0086] Example 2 QPCR sequencing to verify the differential expression of the PLD5 gene

[0087] 1. Large-sample QPCR verification of differential expression of PLD5 gene. According to the sample collection method in Example 1, 50 cases of lung adenocarcinoma paracancerous tissues and 50 cases of lung adenocarcinoma tissues were selected.

[0088] 2. The RNA extraction steps are as described in Example 1.

[0089] 3. Reverse transcription:

[0090] 1) Reaction system:

[0091] Reagent

volume

MgCl 2

2μl

10×RT Buffer

1μl

RNase-free water

3.75μl

dNTP mix

1μl

RNase inhibitor

0.25μl

AMV reverse transcriptase

0.5μl

Oligo-dT Adapter Primer

0.5μl

Experimental sample

1μl

[0092] 2) Reverse transcription reaction conditions

[0093] According to the reverse transcription reaction conditions in RNAPCRKit (AMV) Ver.3.0.

[0094] 42°C~55°C for 60 minutes, 99°C for 2 minutes, ...

Embodiment 3

[0112] Effect of embodiment 3siRNA on PLD5 gene expression

[0113] 1. Cell culture

[0114] Human lung adenocarcinoma cell line A549 was incubated in RPMI1640 medium containing 10% fetal bovine serum and 1% P / S at 37°C, 5% CO 2 , Cultivated in an incubator with a relative humidity of 90%. Change the medium once every 2-3 days, and use 0.25% EDTA-containing trypsin for routine digestion and passage.

[0115] 2. siRNA design

[0116] siRNA sequence against PLD5:

[0117] Negative control siRNA sequence (siRNA-NC):

[0118] The sense strand is 5'-UUCUCCGAACGUGUCACGU-3' (SEQ ID NO.7),

[0119] The antisense strand is 5'-ACGUGACACGUUCGGAGAA-3' (SEQ ID NO.8);

[0120] siRNA1-PLD5:

[0121] The sense strand is 5'-UGAAAUGGUGCAUUUUCUGAA-3' (SEQ ID NO.9),

[0122] The antisense strand is 5'-CAGAAAAUGCACCAUUUCACU-3' (SEQ ID NO.10);

[0123] siRNA2-PLD5:

[0124] The sense strand is 5'-AGUGAAAUGGUGCAUUUUCUG-3' (SEQ ID NO.11),

[0125] The antisense strand is 5'-GAAAAUGCACCAUUU...

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Abstract

The invention discloses a marker for diagnosing and treating lung adenocarcinoma. The PLD5 gene is highly expressed in lung adenocarcinoma tissues, the inhibition of the expression of the PLD5 gene can promote the apoptosis of lung adenocarcinoma cells. The expression level of the PLD5 gene in the lung tissues can be detected to sensitively and specifically diagnose lung adenocarcinoma. The invention also provides a treatment means for lung adenocarcinoma.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a marker for diagnosis and treatment of lung adenocarcinoma, and the marker is PLD5. Background technique [0002] Lung cancer is a disease that seriously endangers human health and life, and its morbidity and mortality have leapt to the top of cancer worldwide. In China, lung cancer is the malignant tumor with the fastest increasing morbidity and mortality rate, and has replaced liver cancer as the primary cause of malignant tumor death. Adenocarcinoma is the main histological type of lung cancer, accounting for more than 50% of all cases, and it is one of the focuses of lung cancer prevention and research. Like other malignant tumors, early detection, early diagnosis, and early treatment are the key to reducing the mortality rate of lung adenocarcinoma. Lung adenocarcinoma mostly originates from the epithelial cells of the small bronchial mucosa, and most adenocarcinomas are located ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886C12Q2600/158G01N33/57423G01N33/57484G01N33/68G01N2333/47
Inventor 杨承刚常鹏宋宏涛
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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