Application of DNAH14 gene in tumor diagnosis and treatment

A gene and gene expression technology, applied in the application field of DNAH14 gene in the diagnosis and treatment of lung adenocarcinoma, can solve the problems of lack of specificity, low sensitivity, drug resistance, etc.

Active Publication Date: 2017-05-10
QINGDAO MEDINTELL BIOMEDICAL CO LTD
View PDF2 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In recent years, with the in-depth study of the molecular mechanism of lung cancer pathogenesis, preliminary results have been achieved in molecular targeted therapy, but there are still some problems such as lack of specificity, low sensitivity, and

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application of DNAH14 gene in tumor diagnosis and treatment
  • Application of DNAH14 gene in tumor diagnosis and treatment
  • Application of DNAH14 gene in tumor diagnosis and treatment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0089] Example 1 Screening for Gene Markers Related to Lung Adenocarcinoma

[0090] 1. Sample collection

[0091] Paracancerous tissue and lung adenocarcinoma tissue samples were collected from 8 cases of lung adenocarcinoma. The tumor tissue specimens of lung adenocarcinoma were collected from the main tumor area, which was located at the junction of the middle and outer 1 / 3 of the tumor mass and normal tissue, excluding the obvious necrosis and calcification of the tumor center and the normal lung tissue around the tumor; the normal lung tissue specimens adjacent to the tumor were obtained from tumor There is no obvious change in the part above 5cm from the edge. All the above specimens were obtained with the consent of the organizational ethics committee.

[0092] 2. RNA sample preparation (using miRNA kit for operation)

[0093] Introduce liquid nitrogen into the mortar, take the tissue obtained above and put it into the mortar, cut it into pieces in liquid nitrogen a...

Embodiment 2

[0115] Example 2 QPCR sequencing to verify the differential expression of DNAH14 gene

[0116] 1. Large-sample QPCR verification of differential expression of DNAH14 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.

[0117] 2. The RNA extraction steps are as described in Example 1.

[0118] 3. Reverse transcription

[0119] 1) Reaction system:

[0120] RNA template 1 μl, random primer 1 μl, double distilled water was added to 12 μl, mixed, centrifuged at low speed, 65 ° C for 5 min, and then cooled on ice.

[0121] Continue to add the following components to the 12 μl reaction solution:

[0122] 5× reaction buffer 4μl, RNase inhibitor (20U / μl) 1μl, 10mM dNTP mixture 2μl, AMV reverse transcriptase (200U / μl) 1μl; mix well and centrifuge;

[0123] 2) Reverse transcription reaction conditions

[0124] 5 minutes at 25°C, 60 minutes at 42°C, and 5 minut...

Embodiment 3

[0141] Example 3 Silencing of DNAH14 Gene

[0142] 1. Cell culture

[0143] 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.

[0144] The cells in the culture flask were digested with trypsin and seeded in a 6-well plate to ensure that the number of cells was 2-8×10 5 cells / well, add cell culture medium. Overnight, observe the cell density the next day, and transfection can be performed when the cell density is above 70%.

[0145] 2. Design of siRNA

[0146] Negative control siRNA sequence (siRNA-NC):

[0147] The sense strand is 5'-UUCUCCGAACGUGUCACGU-3' (SEQ ID NO.7)

[0148] The antisense strand is 5'-ACGUGACACGUUCGGAGAA-3' (SEQ ID NO.8)

[0149] siRNA-1:

[0150] The sense strand is 5'-UCAUAUCUUAAAAGUCUUGGU-...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses an application of a gene related to a lung cancer. Tumor is lung adenocarcinoma, a differential expression of DNAH14 is first discovered to be related to the occurrence and development of the lung adenocarcinoma, significant increase of the DNAH14 in a lung adenocarcinoma patient is verified by virtue of QPCR, and an in-vitro experiment proves that an expression of silent DNAH14 genes can effectively inhibit the proliferation of the lung adenocarcinoma cells, reduce the migration and attack rate of the lung adenocarcinoma cells, prompt that the DNAH14 gene can be used for the identification and diagnosis of the lung adenocarcinoma cells to evaluate the disease risk of the lung adenocarcinoma of relevant people, and also prompt that the DNAH14 gene can be used as a medicine target to treat the lung adenocarcinoma and metastasis thereof.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of DNAH14 gene in the diagnosis and treatment of tumors, in particular to the application of DNAH14 gene in the diagnosis and treatment of lung adenocarcinoma. Background technique [0002] Lung cancer is one of the most common malignant tumors of the respiratory system, with a high degree of malignancy and poor prognosis. According to epidemiological surveys, the incidence of lung cancer in men is higher than that in women, and it is related to factors such as smoking, geography, environment and race. In developed countries such as Europe, North America, and Australia, the incidence of lung cancer has decreased year by year due to the decline in smoking rates in recent years, while the incidence of lung cancer in some Asian countries such as China and countries such as Africa has increased year by year. The incidence of lung cancer is related to many factors. Research da...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12Q1/68G01N33/574A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886C12Q2600/136C12Q2600/158G01N33/57423G01N33/57484
Inventor 边洋李曙光
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap