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Long chain noncoding RNA-HOTAIR molecular marker for colorectal cancer and application of long chain noncoding RNA-HOTAIR molecular marker

A long-chain non-coding technology for colorectal cancer, which can be used in medical preparations containing active ingredients, drug combinations, and microbial determination/examination, etc.

Inactive Publication Date: 2018-11-02
汕头大学医学院附属肿瘤医院
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the unclear diagnosis caused by the level of technology, it is likely that some patients with early colorectal cancer will lose the best treatment opportunities

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 The correlation between HOTAIR and p21

[0020] Tissue specimens were collected from 120 fresh colorectal cancer cases resected by surgery and 120 normal morphological cases adjacent to the cancer, and HOTAIR levels were quantified. Firstly, the HOTAIR siRNA expression vector was constructed and identified in vitro. RT-qPCR was used to detect the expression levels of LncRNA HOTAIR and p21 mRNA in 8 CRC cell lines, CRC tissues and normal tissues adjacent to cancer. The expression levels of LncRNA HOTAIR and p21 and patients were analyzed. Correlation of clinicopathological characteristics (including lymph node metastasis, tumor metastasis, Dukes staging, distant metastasis, histological type and degree of differentiation), Transwell and Scratch experiments detect the migration ability, invasion ability and flow rate of each group of cells transfected with HOTAIR siRNA. Cell apoptosis was detected by cytometry after transfection of HOTAIR siRNA in each group. Cell ...

Embodiment 2

[0022] Example 2 Study the regulatory mechanism of LncRNA-HOTAIR mediated p21 on the invasion and metastasis of colorectal cancer

[0023] Methods: Real-time fluorescence quantitative PCR was used to detect the expression levels of HOTAIR and p21 mRNA in 8 colon cancer cell lines and 120 patients with colorectal cancer in the primary tumor tissues and adjacent tissues, and analyze the relationship between the expression of HOTAIR and p21 and the clinicopathological characteristics of the patients relationship. In addition, M5 cells were divided into blank control group, HOTAIR NC group and HOTAIR siRNA group. Compared with normal human colorectal epithelial cells HIEC, the MTT method detects the cell proliferation of each group; flow cytometry detects the cell cycle and apoptosis rate of each group; Transwell and scratch experiments detect the invasion and migration ability of each group of cells; fluorescence quantitative PCR and Western blot method was used to detect the chang...

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PUM

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Abstract

The invention discloses a long chain noncoding RNA marker for auxiliary diagnosis of colorectal cancer (CRC). Long chain noncoding RNA is HOTAIR; auxiliary diagnosis or prediction of patient conditiondevelopment of the CRC can be realized by detecting expression quantity of the long chain noncoding RNA; differentiation, infiltration depth, proliferation and metastasis conditions of the CRC can bepredicted by monitoring expression conditions of p21 and a value of judging prognosis is realized. Expression level of LncRNA HOTAIR gene can be detected by RT-qPCR, real-time quantitative PCR, immunodetection, in-situ hybridization, chip or a high-throughput sequencing platform; meanwhile, the invention also discloses silencing HOTAIR which can obviously inhibit the proliferation, migration andinvasion abilities of CRC cells, reduce the expression of PCNA, Ki67, cyclin E and CDK2 and medicate expression increase of p21. The invention provides a molecular marker for the CRC and provides theoretical basis for mechanism research and clinical individualized therapy of the CRC; a brand-new treatment strategy can be provided for clinically treating the CRC by monitoring the expression of theLncRNA HOTAIR in the CRC.

Description

Technical field [0001] The present invention relates to the field of biomedicine, in particular to long-chain non-coding RNA markers used to assist in the diagnosis of colorectal cancer and predict colorectal cancer through targeting-mediated up-regulation of p21 expression. Background technique [0002] Colorectal cancer (colorectal cancer, CRC) is a common malignant tumor of the digestive tract. The annual mortality rate of malignant tumors worldwide accounts for about 10%. Although the incidence of colorectal cancer in China is lower than that in Western countries, its incidence is still rising rapidly. Its morbidity and mortality rate ranks 3rd among all malignant tumors in my country, second only to lung cancer and gastric cancer, and its mortality rate ranks 5th, which has seriously affected people's health and quality of life. There are no obvious clinical symptoms in the early stage of colorectal cancer, and there is a lack of specific and sensitive detection methods. Mo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886A61K45/00A61P35/00
CPCA61K45/00A61P35/00C12Q1/6886C12Q2600/178
Inventor 张述耀李从铸林志雄佘玉奇郑洁婷陈钟琳陈晨方翎
Owner 汕头大学医学院附属肿瘤医院
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