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Application of circ-0000423 to preparation or screening of colon cancer drugs

A colon cancer, colorectal cancer technology, applied in the field of biomedicine

Active Publication Date: 2018-10-02
JIANGSU CANCER HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the exploration of colorectal cancer-related circRNAs is just in its infancy, and there is no mechanism involved; therefore, it is necessary to explore the molecular mechanism regulating the invasion and metastasis of colorectal cancer from the perspective of circRNAs

Method used

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  • Application of circ-0000423 to preparation or screening of colon cancer drugs
  • Application of circ-0000423 to preparation or screening of colon cancer drugs
  • Application of circ-0000423 to preparation or screening of colon cancer drugs

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Experimental program
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Effect test

Embodiment 1

[0030] (1) The gene chip was completed by Beijing Kangcheng Company; the human circRNAs / mRNA 4×180K Microarray co-expression chip V.2.0 platform customized by Agilent of the United States was used, covering 4425 circRNAs and 32197 mRNAs; for the original data, Lowess (locally weighted scatterplot smoothing) and other methods to normalize the signal value, then carry out differential gene screening. SAM (Significance Analysis of Microarrays) software was used to analyze the differential genes, and the FDR was controlled within 5%. Using methods such as multiple method and multiple hypothesis testing, the circRNAs and mRNAs with differential expression under two or more conditions were calculated and screened respectively.

[0031] (2) GO functional annotation Gene ontology (Gene ontology, GO) functional annotation is one of the most widely used methods for gene functional annotation. The GO database (www.geneontology.org / ) was created by the Gene Ontology consortium. The GO da...

Embodiment 2

[0036] According to the tissue microarray, 4 transcripts with expression differences of more than 5 times, high abundance and good uniformity were further screened out of 41 circRNAs, of which 3 were up-regulated and 1 was down-regulated (Table 1) ( image 3 A-D).

[0037] Table 1. According to the chip results, 3 up-regulated and 1 down-regulated circRNAs were repeatedly screened

[0038]

[0039] Subsequently, we performed tissue PCR verification on the above four differentially expressed circRNAs in 20 pairs of colorectal cancers:

[0040] Extract total RNA from colon cancer tissue, synthesize cDNA according to the instructions of the TAKARA036A kit, and perform fluorescent quantitative PCR reaction (see Table 2 for primers): according to the principle of experiment repetition and control, set up three replicate wells for each group of samples each time, and set up a negative control compared with blank. Take the 10 μL system as an example according to the instructions:...

Embodiment 3

[0047] The expression of circ-0000423 in colon cancer cell lines HT29 and HCT116 was verified by designing specific back-to-back primers for circ-0000423 junction and performing RT-PCR (see Table 2 for primers of circ-000423). At the same time, in view of the special structure of circRNA and its insensitivity to RNase, RT-PCR successfully identified the existence of circ-0000423 in colorectal cancer cell lines after RNase R treatment ( Figure 4 ); at the same time, searching UCSC shows that: circ-0000423 is located on the PPP1R12A gene of the long arm of chromosome 12, formed by the circularization of exons 1 and 2 of PPP1R12A, with a length of 1138bp, and the sequence is shown in SEQ ID NO.1 ( Figure 5 ).

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Abstract

The invention discloses the application of circ-0000423 to preparation or screening of colon cancer drugs. High expression of circ-0000423 is closely related to staging and differentiated degree degrees and prognosis of colon cancer. The higher the expression degree is, the later the illness is, and the worse the differentiation and prognosis are, which indicates that the circ-0000423 is closely related to malignant activity of colorectal cancer. The circ-0000423 is capable of adsorbing cancer suppressor genes miR-519 to promote invasion and metastasis of the colorectal cancer by a mechanism ceRNA. The circ-0000423 can serve as a therapeutic target for preparing or screening medicines for treating colon cancer or inhibiting invasion and metastasis of the colon cancer. The circ-0000423 canserve as a detection target in preparation of a diagnostic reagent for the colorectal cancer. The circ-0000423 can serve as a detection target in preparation of a reagent for predicting staging and differentiated degree degrees or prognosis of the colorectal cancer.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of circ-0000423 in the preparation or screening of colon cancer drugs. Background technique [0002] Colorectal cancer is the third most common cancer in men worldwide, the second in women, and the fourth deadliest cancer; in 2012, there were 1.4 million new cases of colorectal cancer and 694,000 deaths; the original incidence rate was low of East Asia are rapidly increasing [1-3] . At present, the treatment of colorectal cancer is still dominated by surgery, radiotherapy and chemotherapy, and targeted drugs, but the 5-year survival rate has not been significantly improved; the main cause of treatment failure is cancer invasion and metastasis [4,5] . It is extremely urgent to actively explore the molecular mechanism of invasion and metastasis of colorectal cancer and find new targets for diagnosis and treatment to improve the curative effect and prognosis. [0003] Wit...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886A61K48/00A61K31/713A61P35/00
CPCA61K31/713A61P35/00C12Q1/6886C12Q2600/106C12Q2600/118C12Q2600/178
Inventor 杨柳马大伟
Owner JIANGSU CANCER HOSPITAL
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