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Circ-WHSC1 serving as colorectal cancer diagnostic marker and application thereof

A technology of colorectal cancer and biomarkers, applied in the field of tumor marker medicine, can solve problems that have not been reported, and achieve the effect of inhibiting growth and metastasis

Active Publication Date: 2018-12-07
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of the diagnostic markers for colorectal cancer are linear genes. The Chinese patent application number 2017108397037 reports the application of a circular RNA as a diagnostic biomarker for colorectal cancer. and its circular RNA diagnostic markers for liver metastases have not been reported

Method used

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  • Circ-WHSC1 serving as colorectal cancer diagnostic marker and application thereof
  • Circ-WHSC1 serving as colorectal cancer diagnostic marker and application thereof
  • Circ-WHSC1 serving as colorectal cancer diagnostic marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1c

[0033] Example 1 Identification of circ-WHSC1 and its detection in clinical colorectal cancer specimens

[0034] The mouse colorectal cancer liver metastasis animal model was used to obtain in situ tumors and liver metastatic tumors, and the whole transcriptome sequence was performed. The specific methods are as follows:

[0035] Purchase 4-6 weeks of C57 mice, after anesthesia, 1×10 6 Mouse colorectal cancer cells (CMT-93) were injected under the colorectal mucosa of mice, and the mice were sacrificed 8-10 weeks later; tumors in situ and liver metastases were collected; 3 pairs of samples were sent for whole transcriptome sequencing . Such as Figure 1-4 As shown, figure 1 It is a schematic diagram of the structure of ring-shaped circ-WHSC1; figure 2 The ring in the black box is circ-WHSC1; image 3 Sanger sequencing image of circ-WHSC1 in mouse tissues. In the figure, AAAG and TGTT under the arrow indicate binding sites. The results show that circ-WHSC1 does exist in mouse tiss...

Embodiment 2c

[0043] Example 2 The level of circ-WHSC1 directly affects colorectal cancer tumor growth and metastasis

[0044] 1. Construction of stable cell lines:

[0045] The overexpression plasmid (pLCDH-circ-WHSC1) and siRNA were constructed by Guangzhou Jisai Company. After the identification of the forward transfection was correct, they were repackaged and constructed respectively to obtain the lentivirus (10 7 And 10 9 ). One day before the infection, 100,000 intestinal cancer cells were seeded in a p35 petri dish, and the next day for lentiviral infection (MOI=50), while adding 5μg / mL polybrene (polybrene) to enhance the infection efficiency. After 48 hours of infection, the medium was changed, and the culture was continued for 24 hours so that the genome of the lentivirus was fully integrated into the genome of the colon cancer cell. Passage when the cell plating rate reaches 90%, and at the same time, add 5μg / mL puromycin to the culture medium to screen for stable infected cells. The...

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Abstract

The invention discloses an application of circular RNA WHSC1 in preparation of clinical diagnosis markers in colorectal cancer tumorigenesis and hepatic metastases diseases. The invention further discloses an application of the circular RNA WHSC1 serving as a therapeutic target of the colorectal cancer drug. The invention further discloses a preparation for treating colorectal cancer. The preparation comprises a sequence for specifically interfering the circular RNA WHSC1, wherein the sequence is used for inhibiting the level of the circular RNA WHSC1. Research results show clinical significances of the circ-WHSC1 in diagnosis of colorectal cancer and metastasis and an application in preparation of drugs for treating the colorectal cancer metastasis.

Description

Technical field [0001] The present invention relates to the technical field of tumor marker medicine, in particular to circ-WHSC1 as a diagnostic marker for colorectal cancer and its application. Background technique [0002] With the changes in people's living and eating habits and work and living environment, the incidence and mortality of colorectal cancer (CRC) are increasing year by year around the world. The early onset of CRC is insidious and lacks specific clinical manifestations. Therefore, most patients have already metastasized when they go to the hospital for treatment. The liver is the most important target organ for remote metastasis of CRC. Liver metastasis in CRC patients is the most important cause of death. The 5-year survival rate after liver metastasis drops significantly. Clinically, if CRC and its liver metastasis can be detected and diagnosed early, and comprehensive treatment including radical surgery can be performed in a timely and reasonable manner, th...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 李建明孙丽娜陈良燕
Owner SUZHOU UNIV
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