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Orrhology detection method and use of substrate metal protease MMP11

A MMP11, matrix metal technology, applied in the direction of measuring devices, scientific instruments, instruments, etc., can solve the problem of losing the best treatment period, and achieve the effect of high sensitivity and specificity

Inactive Publication Date: 2008-12-17
BEIJING CANCER HOSPITAL PEKING UNIV CANCER HOSPITAL
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  • Claims
  • Application Information

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Problems solved by technology

Gastric cancer is one of the most common malignant tumors in my country. At present, the diagnosis of gastric cancer mainly relies on imaging and pathology. Most patients have reached the middle and late stages when they are diagnosed, and have lost the best treatment period.

Method used

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  • Orrhology detection method and use of substrate metal protease MMP11
  • Orrhology detection method and use of substrate metal protease MMP11
  • Orrhology detection method and use of substrate metal protease MMP11

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Embodiment Construction

[0016] The specific implementation manner of the present invention will be described below in conjunction with the accompanying drawings.

[0017] material method

[0018] 1. Cell Culture

[0019] Gastric cancer cell lines BGC823, MGC803, SGC7901, PAMC82, MKN45, SNU1, SNU5, SNU16, RF1, RF48 were cultured in DMEM medium with 5% fetal bovine serum, and AGS and N87 were cultured in DMEM medium with 10% fetal bovine serum. The base contains penicillin at a final concentration of 100,000units / L and streptomycin at 100mg / L, in 5% CO 2 Incubate at 37°C.

[0020] Second, RT-PCR

[0021] Take 5.0 μg total RNA, add 0.5 μl Oligo(dT), denature at 70°C for 10 minutes, then add 6.0 μl 5×First Strand Buffer, 2.0 μl 2.5mM dNTPs mix, 0.5 μl RNasin (40U / μl) and 1.0 μl MMLV ( 200U / μl), mix well and incubate at 37°C for 1 hour, and finally stop the reaction at 70°C for 15 minutes. The total volume of the PCR reaction system is 20 μl, including 10×PCRBuffer, 2.5mM dNTPs mix, Taq DNA polymeras...

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Abstract

The invention discloses a method for serologically detecting Matrix metalloproteinase 11 (MMP11) and the application thereof. The gene chip technology and the bioinformatics method are adopted to obtain stomach cancer gene expression spectra, and by taking the MMP11 gene for example, the expression characteristics of the gene in the tumor cell lines and tissues are verified at the levels of mRNA and protein. At the same time, a detection kit for the MMP11serum is established by utilizing the technique of the enzyme linked immuno sorbent assay (ELISA), and the expression level of the MMP11 protein in the serum of stomach cancer patients is detected. The level of the MMP11 protein in the serum of stomach cancer patients is obviously higher than that of a non-cancer comparison group (p is lower than 0.001), and the same tendency is also found in the breast cancer, the colon cancer / rectum cancer and the lung cancer. In the serum detection to the stomach cancer patients, the sensitivity of the MMP11 is higher than that of a tumor molecule marker such as CEA, CA199, CA72.4, CA242, etc., and MMP11 and CA199 have good correlation (p is equal to 0.017). Through the analysis of the clinic pathological data, the level of the serum MMP11 has the obvious correlation (p is equal to 0.009) with the cancerometastasis state of the stomach cancer patients. The MMP11 is possible to become a novel tumor serum marker for the diagnosis and the prognostic judgement of the tumors.

Description

technical field [0001] The present invention relates to a variety of cell and molecular biology techniques, including RT-PCR, immunohistochemistry, Western blot, cell transfection and animal experiments, and enzyme linked immunosorbent assay (enzyme linkedimmuno sorbent assay, ELISA) technology. In particular, it relates to a serological detection method and application of matrix metalloproteinase MMP11. Background technique [0002] Malignant tumors have become an important cause of human death, and about 7 million people die of cancer in the world every year. Gastric cancer is one of the most common malignant tumors in my country. At present, the diagnosis of gastric cancer mainly relies on imaging and pathological methods. Most patients have reached the middle and late stages when they are diagnosed, and have lost the best treatment period. Therefore, finding tumor molecular markers is a key issue for early diagnosis and effective prevention and treatment of tumors. Amo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/564G01N33/543
Inventor 吕有勇杨艳红邓华张青云肖白耿排力
Owner BEIJING CANCER HOSPITAL PEKING UNIV CANCER HOSPITAL
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