Non-small cell lung cancer (NSCLC) marker and its application

A technology of non-small cell lung cancer and markers, applied in the direction of drug combination, genetic material composition, biochemical equipment and methods, etc., can solve the problems of no literature reports

Inactive Publication Date: 2012-01-18
THE AFFILIATED HOSPITAL OF SOUTHWEST MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the expression of STAT3 in the tissues and blood of lung cancer patients, especially non-small cell lung cancer patients, and its feasibility as a new diagnostic marker for lung cancer, as well as its related applications, have not been reported in the literature.

Method used

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  • Non-small cell lung cancer (NSCLC) marker and its application
  • Non-small cell lung cancer (NSCLC) marker and its application
  • Non-small cell lung cancer (NSCLC) marker and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0105] Example 1: Expression of STAT3 protein in human lung adenocarcinoma cell A549 and normal human embryonic lung cell MRC-5

[0106] The results of immunohistochemical staining showed that the positive expression was brownish-yellow or brown granular and deposited on the positive site. The positive expression signal of STAT3 protein in human lung adenocarcinoma cell A549 is mainly located in the cytoplasm, and a few are located in the nucleus, showing obvious brownish yellow or brown granular staining (see attached figure 1 ). In the normal human embryonic lung cell MRC-5, it is also weakly expressed in the cytoplasm, showing light yellow coloration (see attached figure 2 ). The t-test was performed between the MOD values ​​expressed in human lung adenocarcinoma cell A549 and normal human embryonic lung cell MRC-5, P=0.006 (P<0.01), and the difference was statistically significant, suggesting that STAT3 protein was in The expression level in human lung adenocarcinoma c...

Embodiment 2

[0110] Example 2: Correlation between the expression of STAT3 protein and CEA protein and CA125 protein in human lung adenocarcinoma cell A549

[0111] 2.1 Expression of CEA protein in human lung adenocarcinoma cell A549 and normal human embryonic lung cell MRC-5

[0112] The positive expression signal of CEA protein in human lung adenocarcinoma cell A549 is mainly located in the cytoplasm and (or) the cell membrane, showing brownish-yellow or yellowish-brown granular staining (see attached image 3 ); normal human embryonic lung cell MRC-5 was also expressed to a weaker degree in the cytoplasm, showing light yellow coloration (see attached Figure 4 ). The MOD value of this protein expressed in human lung adenocarcinoma cell A549 and normal human embryonic lung cell MRC-5 was tested by t test, P=0.000 (P Figure 7 ,table 3).

[0113] Table 3 CEA protein expression in A549 and MRC-5 cells ( ±s)

[0114]

[0115] Note: a P<0.01 vs MRC-5

[0116] 2.2 Expression of CA12...

Embodiment 3

[0127] Example 3: Expression of STAT3 mRNA in human lung adenocarcinoma cell A549 and normal human embryonic lung cell MRC-5 In this experiment, the SYBR Green I fluorescent dye method was selected, and the melting curve analysis step was added to the experimental design, which was analyzed by Bio-Rad IQ5 Software analysis. Melting curves showed that each gene had a single product peak (such as Figure 8-1 , 8-2, 8-3 and 8-4), no non-specific products and primer-dimers were generated, indicating that the specificity of the primers was good, and the experimental results could accurately reflect the initial concentration of the sample.

[0128] To establish a standard curve: take a certain amount of cDNA PCR product template, and carry out 10 3 ~10 9 Dilute to obtain 7 concentration gradients, and then take 2 μl of dH 2 O as zero tube. At the same time, real-time PCR amplification was performed to establish a standard curve. The Ct value of the standard PCR product dilute...

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Abstract

Belonging to the field of biotechnology, the invention discloses a non-small cell lung cancer (NSCLC) marker, which is STAT3, and also can include CEA, CA125 and CYFRA21-1. The invention confirms the high expression of STAT3 in peripheral blood and serum, and discloses application of STAT3 in preparation of NSCLC diagnostic reagents by the inventor. Specifically, by making use of a genetic recombination technology and targeting at the gene coding region of STAT3, an eukaryotic expression vector PSUPER-STAT3 able to transcribe in vivo and generate small interference RNA (siRNA) is constructed successfully, and is transfected into an eukaryotic cell, thus laying a foundation for a further experimental study on lung cancer RNAi (RNA interference) and antitumor gene therapy. Utilization of the RNAi technology can effectively inhibit the gene expression of STAT3, induce cell apoptosis, and establish the base for molecular mechanism research and gene therapy of lung cancer, thus providing application of the marker provided in the invention in preparing related targeted medicines.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a tumor marker and its application. Background technique [0002] Lung cancer is a common malignant tumor of the respiratory system and is considered the number one killer of cancer. At present, the 5-year survival rate of lung cancer in my country is only 10%, and only 15% of patients have limited lesions when they are diagnosed, and most of them have lost the chance of surgery when they are diagnosed, so the prognosis is extremely poor and the mortality rate is extremely high. [0003] The high mortality rate of lung cancer is mainly due to the inability to achieve early diagnosis. Therefore, early diagnosis of lung cancer is the most effective measure to timely standardize treatment and improve its prognosis. At present, imaging and fiberoptic bronchoscopy are the main means of diagnosing lung cancer. Compared with the two, the detection of tumor markers has the advantages of sim...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/574A61K48/00A61P35/00
Inventor 王鸿程宋本艳蓝秀王超辛彩霞王传辉
Owner THE AFFILIATED HOSPITAL OF SOUTHWEST MEDICAL UNIV
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