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Tumor marker AQUAPORIN2 protein and application thereof

A technology of AQUAPORIN2 and tumor markers, applied in the field of tumor detection and molecular targeted therapy

Active Publication Date: 2021-03-23
NANJING ANJI BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Aiming at the problem that existing head and neck squamous cell carcinoma and other malignant tumors lack closely related biomarkers, the present invention provides a tumor marker AQP2 protein, and successfully applies it in tumor detection and treatment, by combining bioinformatics means, Clinical tumor samples and biological function experiments have discovered new biomarkers that are closely related to the occurrence, development and metastasis of head and neck squamous cell carcinoma, kidney cancer and prostate cancer

Method used

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  • Tumor marker AQUAPORIN2 protein and application thereof
  • Tumor marker AQUAPORIN2 protein and application thereof
  • Tumor marker AQUAPORIN2 protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Microarray analysis of expression profile of AQP2 in different human tumor tissues and paracancerous tissues

[0048]The Tumor Genome Atlas (TCGA) project is a project jointly launched by the National Cancer Institute (NCI) and the National Human Genome Research Institute (NHGRI) in 2006. It uses large-scale sequencing-based genome analysis technology to conduct large-scale research on 36 cancers. In experiments, TCGA Genome Analysis Center (GCC) compares tumors and normal tissues to find gene mutations, amplifications or deletions associated with each cancer or subtype. To help understand the molecular mechanism of cancer and improve people's scientific understanding of the molecular basis of cancer pathogenesis.

[0049] Using the TCGA standard method to download the full gene expression profile data and clinical information of 36 tumors and their paracancerous tissues, using R language (version 3.1.1) software to filter out tumor types that do not contain AQP2 expres...

Embodiment 2

[0056] In this example, the expression of AQP2 in tumor cells and normal epithelial cells was detected by fluorescent quantitative PCR method.

[0057] 1. Materials

[0058] Head and neck squamous cell carcinoma cells SCC4 and human normal oral epithelial cells HIOEC, kidney cancer cells 786-O and human renal epithelial cells HEK293T, prostate cancer cells DU145 and human normal prostate epithelial cells RWPE-1; the above cells were purchased from ATCC cell bank in the United States.

[0059] 2. Method

[0060] 2.1 Extraction of total RNA from tumor cells and normal epithelial cells

[0061] The above six kinds of cells were respectively stored at 37°C and 5% CO 2 cultured in an incubator, cultured to a density of 90%, digested with trypsin and collected, resuspended the cells in the culture medium and counted them under a microscope, and adjusted the cell concentration to 5×10 5 cells / mL, then inoculate the cell suspension with adjusted concentration into 6-well plates, 2 ...

Embodiment 3

[0073] In this example, the expression of AQP2 protein in tumor cells and normal epithelial cells was detected by immunoblotting.

[0074] Digest and collect the six kinds of cells in Example 2 when the growth density reaches 90%, resuspend the cells in culture medium for expansion culture, then collect the cells at 80% confluence, discard the supernatant after centrifugation, and rinse with PBS Twice, discard the supernatant. Add RIPA lysate and lyse on ice for 20 min. The supernatant was collected by centrifugation at 12000 g for 10 min. Add 1X SDS sample buffer, mix well by pipetting and boil for denaturation for 5min. Total proteins were separated by 10% SDS-PAGE gel and then transferred to PVDF membrane. 5% BSA was blocked at room temperature for 2 hours, incubated with AQP2 antibody (abcam) overnight at 4°C, and washed 3 times with TBST. The secondary antibody was incubated at room temperature for 1 h, and washed 3 times with TBST. ECL ultra-sensitive chemiluminesce...

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Abstract

The invention discloses a tumor marker AQUAPORIN2 protein and application thereof, and belongs to the field of tumor detection and molecular targeted therapy. The invention comprises the application of the AQUAPORIN2 protein in the following functions: (1) serving as a new tumor marker for early diagnosis and prognosis effect judgment of malignant tumors including head and neck squamous carcinoma,kidney cancer and prostate cancer; (2) inhibiting tumor cell proliferation; (3) inhibiting tumor cell migration; and (4) inhibiting growth of an animal transplantation tumor model. The targeted therapy with the AQUAPORIN2 protein as a biomarker provides a new idea for treatment of malignant tumors such as head and neck squamous carcinoma, kidney cancer and prostate cancer.

Description

technical field [0001] The invention belongs to the field of tumor detection and molecular targeted therapy, and more specifically relates to a transmembrane protein AQUAPORIN 2 (abbreviated as AQP2 in the text) and its application. Background technique [0002] Tumor is the most serious disease that endangers human health. Studies have found that the occurrence of tumors is a complex process of gradual accumulation of gene mutations. The development of modern medical technology and molecular biology has brought tumor treatment into an era of individualization and greatly increased the remission rate of tumor treatment. Therefore, finding specific targets for early diagnosis, treatment and prognosis of tumors is the key bottleneck restricting the clinical efficacy of tumors. [0003] Head and neck cancers include neck tumors (thyroid tumors, etc.), otolaryngology tumors (larynx cancer, nasopharyngeal cancer, paranasal sinus cancer, etc.) % of head and neck tumors are squam...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/08A61K45/00A61P35/00A61P35/04C12Q1/6886G01N33/574
CPCA61K38/08A61K45/00A61P35/00A61P35/04C12Q1/6886G01N33/57438G01N33/57434G01N33/574C12Q2600/158G01N2800/60G01N2800/52G01N33/57492A61K38/1709C07K14/705C12Q2600/106
Inventor 徐寒梅李梦玮
Owner NANJING ANJI BIOLOGICAL TECH CO LTD
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