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Application and kit of gene ikzf3 in preparation of lung cancer diagnostic kit

A diagnostic kit and gene technology, applied in the fields of molecular biology and oncology, can solve the problems of low specificity and imperfection

Active Publication Date: 2016-08-31
TIANJIN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, tumor markers that have been used clinically include alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), etc., but generally speaking, they are still not perfect and the specificity is not high. There is still an urgent need for more and better, Discovery / invention and clinical application of tumor markers and detection kits with tumor type specificity

Method used

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  • Application and kit of gene ikzf3 in preparation of lung cancer diagnostic kit
  • Application and kit of gene ikzf3 in preparation of lung cancer diagnostic kit
  • Application and kit of gene ikzf3 in preparation of lung cancer diagnostic kit

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Experimental program
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Effect test

Embodiment 1

[0035] The preparation of embodiment 1 DNA standard substance

[0036] Total RNA was extracted from the human small cell lung cancer cell line H209, and total cDNA was obtained by RT. The 1008bp GAPDH gene ORF was amplified by PCR using primers 7 and 8, and the 1530bp IKZF3 gene ORF was amplified by PCR using primers 9 and 10, respectively. After being digested with EcoRI, it was connected into the pUC19 plasmid that was also digested with EcoRI, and transformed into competent cells E.coli DH5α, and the clones were selected and sequenced to confirm their correctness, and pUC19-GAPDH plasmid and pUC19-IKZF3 were constructed plasmid. Amplify and extract the two plasmids, measure the concentration, mix the two evenly, and dilute to obtain DNA standards with five concentration gradients. The five concentration gradients are 10pg / ml, 100pg / ml, 1ng / ml, and 10ng / ml respectively. and 100ng / ml, said concentration is equivalent to the concentration of gene GAPDH molecule (1008bp) and g...

Embodiment 2

[0038] Example 2 IKZF3 is specifically expressed in various types of lung cancer tissues, but not expressed in paracancerous tissues by immunohistochemistry

[0039] We took 40 paracancerous tissues of lung cancer patients, 40 lung squamous cell carcinoma tissues, 40 lung adenocarcinoma tissues and 40 small cell lung cancer tissues from our own lung cancer patient tissue sample bank, and detected these tissues by immunohistochemistry The expression of IKZF3 in . It was found that the expression of IKZF3 was not detected in the paracancerous tissues, but the expression of IKZF3 was detected in squamous cell carcinoma, adenocarcinoma and small cell lung cancer. We semi-quantitatively analyzed the expression of IKZF3 in lung paracancerous tissues, squamous cell carcinoma tissues, adenocarcinoma tissues, and small cell lung cancer tissues through the staining depth of cells and the number of positively stained cells. Rated on a scale of 0-8.0. The results found that IKZF3 was no...

Embodiment 3

[0041] Example 3 Research on the relationship between the relative expression of IKZF3 and GAPDH in lung cancer tissues and the prognosis of lung cancer patients

[0042] We screened 200 cases of lung cancer tissues with clear case follow-up records from our own lung cancer patient tissue sample bank, and detected the relative expression of IKZF3 relative to GAPDH in these lung cancer tissues by using the diagnostic kit provided by the present invention and its use method, Set the molecular concentration value of GAPDH in each group of samples as x, and the molecular concentration value of IKZF3 as y, and calculate the ratio of y / x to be the relative expression of IKZF3 gene. We analyzed the relationship between y / x and patient survival time. Correlation. Statistical analysis of the data showed that the relative expression of IKZF3 in lung cancer tissues was strictly negatively correlated with the survival time of patients (P Figure 4 .

[0043] Specific steps are as follows:...

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Abstract

The invention provides an application of a gene IKZF3 in preparing a lung cancer diagnosis kit and the kit. The kit is composed of five parts, including a Real-Time PCR (Polymerase Chain Reaction) primer, a DNA (Desoxvribose Nucleic Acid) standard substance, an RNA (Ribose Nucleic Acid) extraction system, an RNA reverse transcription system and a Real-Time PCR system. The kit is used for analyzing the tumor tissue of a clinical lung cancer patient, detecting the relative expression quantity of the target gene IKZF3 relative to a reference gene GAPDH (reduced glyceraldehyde-phosphate dehydrogenase) in a sample and thus judging the prognosis of the patient.

Description

technical field [0001] The invention belongs to the fields of molecular biology and oncology, and relates to nucleic acid detection reagents and their application in tumor prognosis. Background technique [0002] In recent years, the incidence of malignant tumors has increased year by year, and it has become one of the main factors leading to clinical death of patients. Tumor markers (Tumor Marker TM) have great practical value in tumor screening, diagnosis, prognosis, evaluation of treatment efficacy, and follow-up of high-risk groups. It is one of the key issues in current oncology research. At present, tumor markers that have been used clinically include alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), etc., but generally speaking, they are still not perfect and the specificity is not high. There is still an urgent need for more and better, Discovery / invention and clinical application of tumor markers and detection kits with tumor type specificity. [0003] IKZF...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/118C12Q2600/158
Inventor 刘喆马振毅李西川徐昭
Owner TIANJIN MEDICAL UNIV
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