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Applications of CTx to preparation of medicaments for diagnosing colorectal cancer

A technology for colorectal cancer and rectal cancer, which can be used in disease diagnosis, biological testing, biological material analysis, etc., and can solve problems such as unreported clinical value.

Active Publication Date: 2014-01-22
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in addition to being used as a standard for judging tumor bone metastasis, the clinical value of CTx in other tumor development processes and the diagnosis and prognosis of colorectal cancer has not been reported.

Method used

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  • Applications of CTx to preparation of medicaments for diagnosing colorectal cancer
  • Applications of CTx to preparation of medicaments for diagnosing colorectal cancer
  • Applications of CTx to preparation of medicaments for diagnosing colorectal cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Collection of colorectal cancer patient samples and normal samples at different stages

[0038] From May 2009 to May 2010, we collected tissue and serum samples from 91 patients with colorectal cancer (the patients had been diagnosed with colorectal cancer by histopathological examination, and according to the International Union Against Cancer (UICC) and The TNM (tumor-node-metastasis) staging method jointly developed by the American Cancer Society (AJCC) determined the pathological stage, including 21 patients with stage I, 41 patients with stage II, 22 patients with stage III, and 7 patients with stage IV. laparoscopic surgery), and collected 33 healthy human serum samples without digestive tract diseases and inflammatory diseases. All patients were followed up every 3 months in the first 2 years after surgery, followed up every 6 months in the third year after surgery, and stopped at the 41st month after surgery, and the time points of recurrence or death...

Embodiment 2

[0042] Example 2: Expression of Type I Collagen at RNA and Protein Levels in Different Stages of Colorectal Cancer Tissues

[0043] 1. Tissue RNA extraction and real-time fluorescent quantitative PCR (Realtime PCR) detection:

[0044] We extracted the total mRNA of tumor tissue and corresponding normal colorectal tissue, and detected the mRNA expression level of type I collagen respectively. The total mRNA of the tissue was extracted with the Trizol kit from Invitrogen, and 20ml of cDNA was synthesized by reverse transcription from 2 mg of RNA using the PrimeScript RT reagent Kit from Takara, and the 7500 real-time fluorescent quantitative PCR instrument from Applied Biosystems and the corresponding SYBR Green PCR were used. The Master Mix kit was used for real-time fluorescent quantitative PCR (2-△△Ct analysis method was used to analyze the expression abundance, and GAPDH was used as an internal reference gene). The primer sequences used in the PCR reaction are shown in Tabl...

Embodiment 3

[0059] Example 3: ELISA detection of type I collagen degradation index CTx and random degraded small fragment COL1 in the serum of normal and different stages of colorectal cancer patients:

[0060] 1. ELISA detection kit and method

[0061] The levels of COL1 and CTx in serum can be detected by ELISA using a commercial kit (Uscn Life Science&Technology Co., Houston, TX, USA), and serum CTx can be detected by linear EKAHDGGR peptide (fragment of α1 chain carboxy-terminal peptide of type I collagen). Competitive inhibition ELISA detection of specific monoclonal antibodies, in short, is biotin-labeled synthetic CTx peptides and unlabeled CTx in serum and pre-coated specific CTx antibodies for competitive inhibition reactions, each reaction Add avidin horseradish peroxide into the microwells and incubate. For the specific detection method, refer to the kit instructions. After adding the substrate solution, the color of the reaction mixture was negatively correlated with the conc...

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Abstract

The invention relates to the technical field of biology, and especially relates to applications of CTx to preparation of medicaments for diagnosing colorectal cancer, wherein CTx is a fragment at the carboxyl telopeptide of an extracellular matrix component type I collagen. The invention provides applications of CTx to prepare and screen medicaments for diagnosing colorectal cancer and screen medicaments for treating colorectal cancer. Colorectal cancer tissues and serum clinic specimens are selected for researching the clinic value of type I collagen on generation, development, staging prognosis and the like of colorectal cancer. Through a large number of experiments, inventors of the invention discover that the expression level change of CTx in serums of colorectal cancer patients at different stages is in positive correlation with pathology staging, and compared with conventional alimentary canal tumour oncofetal antigens, CTx is capable of better differencing the metastatic colorectal cancer patients and the non-metastatic colorectal cancer patients. Additionally, the inventors also discover that CTx is correlated with 3-year tumor-free survival of the colorectal cancer patients, and the high expression of CTx indicates poor tumor-free survival and prognosis of the colorectal cancer patients.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the use of a carboxyl terminal peptide CTx of type I collagen, an extracellular matrix component, in the preparation of drugs for diagnosing colorectal cancer. Background technique [0002] Collagen, as one of the main components of the extracellular matrix (ECM), plays an important role in maintaining the structural stability of the ECM in most connective tissues, and is associated with many fibrous diseases such as liver fibrosis (Lai, K.K.,Shang,S.,Lohia,N.,etc.Extracellular matrix dynamics in hepatocarcinogenesis: a comparative proteomics study of PDGFC transgenic and Pten null mouse models.PLoS Genet.,2011,7(6):e1002147.). Recent reports also point out that collagen is also very important for the invasion and metastasis of tumor cells (Sund, M., Kalluri, R.Tumor stroma derived biomarkers in cancer. Cancer Metastasis Rev., Jun, 2009,28(1-2): 177 -83.). At present, it has been r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6893G01N33/94G01N2800/065G01N2800/52G01N2800/56
Inventor 邹霞张延杨芳冯波郑民华
Owner SHANGHAI JIAO TONG UNIV
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