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Micro RNA biomarker for predicting early non-metastatic colorectal cancer prognosis and detection method

A colorectal cancer, non-metastatic technology, applied in the field of biomedicine, can solve the problems of unreported miRNA and miRNA differences

Active Publication Date: 2015-06-03
上海兰卫医学检验所股份有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the above results are based on the whole tissue, and the previous research of the inventors of the present application found that there are differences in the expression profiles of miRNAs between different types of cells (Wang, S. (2010) BMC Genomics, 11: p.163) , so far, no miRNAs associated with the prognosis of early non-metastatic colorectal cancer have been reported

Method used

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  • Micro RNA biomarker for predicting early non-metastatic colorectal cancer prognosis and detection method
  • Micro RNA biomarker for predicting early non-metastatic colorectal cancer prognosis and detection method
  • Micro RNA biomarker for predicting early non-metastatic colorectal cancer prognosis and detection method

Examples

Experimental program
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Embodiment 1

[0151] Example 1: Patient Data

[0152] In the discovery experimental group, a total of 8 cases of large intestine paraffin tissue specimens archived in Huashan Hospital Affiliated to Fudan University in 2005 with five-year postoperative survival data were used. The patient's prognosis data came from the Shanghai Center for Disease Control. Among them, there were 4 cases of five-year survival after operation and four cases of death due to recurrence or metastasis of colorectal cancer within five years after operation. All participating patients had informed consent to the scientific research. All procedures for collecting specimens were approved by the Institutional Review Board of Shanghai Medical College.

[0153] In the verification experiment, a total of 56 colorectal cancer patients' paraffin tissue samples were used. The paraffin tissue samples of large intestine archived in Huashan Hospital Affiliated to Fudan University in 2005 were used. The patient's prognosis dat...

Embodiment 2

[0158] Embodiment 2: sample preparation

[0159] In discovery experiments, laser capture microdissection was optionally performed on each cancer sample to specifically isolate tumor cell populations (approximately 200,000 cells). Briefly, a clear transfer film is applied to the surface of a tissue section or sample. Under a microscope, thin tissue sections mounted on glass slides are viewed and cell populations are identified for isolation. When the selected cell is in the center of the observed field of view, a near IR laser diode integral with the microscope optics is activated. A pulsed laser beam activates spots on the transfer membrane, causing the membrane to fuse with the underlying selected cells. The transferred membrane with bound cells is then stripped from the thin tissue section (for review see e.g. Emmert-Buck, M.R. et al. (1996). Science 274, 998-1001; Espina, V. et al. (2007) Expert Rev. Mol. Diagn. 7, 647-657). Cryostat sections were prepared and a laser...

Embodiment 3

[0162] Example 3: chip data

[0163] In experiments, qualitative analysis of (differentially) expressed miRNAs in specific samples can optionally be performed using the Agilent miRNA microarray platform (Agilent Technologies, Santa Clara, CA, USA) according to the manufacturer's instructions. The chip contains 723 human microRNAs selected from Sanger database v.10.1. The amount of total RNA required for each sample was 100ng, and the Cy3 staining marker was incorporated. Chip scanning is through XDR Scan (PMT100, PMT5). The specific operation steps of labeling and hybridization are detailed in Agilent miRNA chip platform system. Raw data obtained for single-color (CY3) hybridization were normalized by applying the Quantile method and using GeneSpring GX10 software known in the art (Agilent Technologies, Santa Clara, CA, USA).

[0164] Unpaired t-test test was used to distinguish the statistical significance of differential expression of prognostic microRNAs in early non-m...

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Abstract

The invention belongs to the technical field of biomedicine and relates to a micro RNA biomarker for predicting early non-metastatic colorectal cancer prognosis and a detection method. The early non-metastatic colorectal cancer refers to Dukes' A and Dukes' B glandular cancers. Particularly, the invention relates to a diagnostic kit for identifying molecular markers of one or more mammalian target cells for prognosis of different early non-metastatic colorectal cancers. The kit comprises multiple nucleic acid molecules, a miRNA sequence is encoded by each nucleic acid molecule, one or more in the multiple nucleic acid molecules have differential expressions in target cells and one or more control cells, and one or more nucleic acid molecules with differential expressions represent nucleic acid expression characteristics together, wherein the nucleic acid expression characteristics refer to indications for identifying prognosis of different early non-metastatic colorectal cancers. The invention further relates to a corresponding method for preventing or treating the disease by using the biomarker and detection method and a pharmaceutical composition.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and relates to a biomarker and a detection method, in particular to a microRNA (microRNA) biomarker and a detection method for reliably predicting early non-metastatic colorectal cancer in colorectal cancer surgically resected tissues method. Among them, early non-metastatic colorectal cancer means Dukes'A and Dukes'B adenocarcinoma. Background technique [0002] The prior art discloses that cancer originates in the epidermis and develops through a multistep transformation: from normal cells to dysplasia and finally to malignant transformation to neoplastic cells, which invade surrounding tissues and have the capacity to metastasize. Colorectal cancer (also known as colorectal cancer (CRC)) is a major cancer type that undergoes this neoplastic development. [0003] Colorectal cancer includes tumors that grow in the colon, rectum, and appendix. Studies have shown that colorectal cancer is t...

Claims

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Application Information

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IPC IPC(8): C12Q1/68A61K48/00A61P35/00
CPCA61K31/7088C12Q1/6886C12Q2600/118C12Q2600/158C12Q2600/178
Inventor 王漱阳朱虹光周春仙吴莹
Owner 上海兰卫医学检验所股份有限公司
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