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Application of CLIC3 as lung adenocarcinoma diagnosis and treatment target

A lung adenocarcinoma and product technology, applied in the fields of treatment, prognosis prediction, and tumor diagnosis, can solve problems such as low diagnostic efficiency and failure to meet clinical needs

Active Publication Date: 2017-08-11
FOURTH HOSPITAL OF HEBEI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the only markers that have been used in the clinical auxiliary diagnosis of lung cancer are protein markers, such as CEA, CA125, and CYFRA21-1. Although the above protein markers have been used in the auxiliary diagnosis of lung cancer, their diagnostic efficiency is low and cannot meet clinical needs.

Method used

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  • Application of CLIC3 as lung adenocarcinoma diagnosis and treatment target
  • Application of CLIC3 as lung adenocarcinoma diagnosis and treatment target
  • Application of CLIC3 as lung adenocarcinoma diagnosis and treatment target

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1 Screening of differentially expressed genes

[0066] 1. Materials:

[0067] The surgically resected cancer tissues and paracancerous tissues of 8 patients with primary lung adenocarcinoma were used as experimental samples. All cancer tissues were pathologically confirmed as lung adenocarcinoma. All patients with primary lung adenocarcinoma had not received radiotherapy and chemotherapy before operation, and the clinical data of all cases were complete.

[0068] 2. Obtaining tissue RNA

[0069] Total RNA was extracted from tissue samples, and the concentration and purity of the extracted RNA were detected by Nanodrop2000, the integrity of RNA was detected by agarose gel electrophoresis, and the RIN value was determined by Agilent2100. The total amount of RNA required for a single library construction is 5ug, the concentration is ≥200ng / μL, and the OD260 / 280 is between 1.8 and 2.2.

[0070] 3. Remove rRNA

[0071] A part (>24%) of the long non-coding RNAs i...

Embodiment 2

[0095] Example 2 Large sample verification screened out differentially expressed genes

[0096] Based on the results of the previous high-throughput transcriptome deep sequencing, we selected the CLIC3 gene for verification according to the size of the P value.

[0097] 1. Sample collection

[0098] According to the method of Example 1, 45 cases of lung adenocarcinoma tissues and corresponding paracancerous tissues were collected.

[0099] 2. Validation at the mRNA level

[0100] 2.1 Extract tissue RNA

[0101] Step is with embodiment 1.

[0102] 2.2 Reverse transcription

[0103] Reverse transcription using Primescript 1 st strand cDNA synthesis kit kit, the operation steps are as follows:

[0104] (1) Add the following reaction liquid in the microcentrifuge tube, as shown in Table 1:

[0105] Table 1 Reaction liquid

[0106]

[0107]

[0108] (2) Incubate at 70°C for 5 minutes, then rapidly cool to 4°C;

[0109] Add the following reagents into a microcentrifu...

Embodiment 3

[0133] Example 3 CLIC3 Gene Overexpression

[0134] 1. Plasmid construction

[0135] Amplification primers were designed according to the coding sequence of the CLIC3 gene, and the design of primers is well known to those skilled in the art. The coding sequence of the full-length CLIC3 gene was amplified from the cDNA library of adult fetal brain (clontech company, catalog number: 638831), the above cDNA sequence was inserted into the eukaryotic cell expression vector pcDNA3.1, and the obtained recombinant vector pcDNA3.1 was connected -CLIC3 was used in subsequent experiments.

[0136] 2. Culture and transfection of lung adenocarcinoma cells

[0137] 2.1 Cell culture

[0138] The lung adenocarcinoma cell line A549 was cultured in RPMI1640 medium and 10% fetal bovine serum.

[0139] 2.2 Cell transfection

[0140] (1) The day before transfection, 0.5-2*10 5 Tumor cells were suspended in 500 μl of antibiotic-free medium and seeded into 24-well culture plates.

[0141] (2)...

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Abstract

The invention discloses a CLIC3 gene which is a lung adenocarcinoma diagnosis and treatment target. By detecting the content of the CLIC3 gene and expression products thereof in the lung tissue of a testee, whether the testee has lung adenocarcinoma or whether the testee is likely to develop lung adenocarcinoma can be judged, and the reactivity of a patient to drug therapy and relapse and prognosis conditions of the patient can be judged. Furthermore, by studying the proliferation index of lung adenocarcinoma cells cultured in vitro, it is proved that the CLIC3 gene and expression products thereof can serve as a drug target for treating lung adenocarcinoma.

Description

technical field [0001] The present invention relates to the fields of tumor diagnosis, treatment, and prognosis prediction. More specifically, the present invention relates to a method for tumor diagnosis and prognosis prediction by means of detecting CLIC3 abnormality; and a tumor therapeutic agent for activating CLIC3 gene or protein. Background technique [0002] Lung cancer is a major disease that seriously threatens human life and health. In 2011, the World Health Organization (WHO) International Cancer Research Agency IARC released the results of 2008 global statistical data, showing that lung cancer is the malignant tumor with the highest incidence and mortality. Every year, there are 1.608 million new cases and 1.377 million deaths worldwide, accounting for 12.7% and 18.2% of all malignant tumors. According to statistics released by the American Cancer Society, it is estimated that in 2012, the number of new lung cancer cases in the United States will reach 226,000,...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886C12Q2600/158G01N33/57423G01N33/57484G01N33/68
Inventor 田子强朱永刚吕会来
Owner FOURTH HOSPITAL OF HEBEI MEDICAL UNIV
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