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Application of CircRNA PVT1 and peptide fragment in tumor growth prediction, metastasis prediction, prognosis evaluation and treatment

A technique for tumor metastasis and prognosis assessment, used in the fields of biotechnology and medicine

Pending Publication Date: 2020-04-24
GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there are no reports on the use of proteins translated from circular RNAs as tumor markers or studies on tumor metastasis, prognosis, and treatment at home and abroad.

Method used

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  • Application of CircRNA PVT1 and peptide fragment in tumor growth prediction, metastasis prediction, prognosis evaluation and treatment
  • Application of CircRNA PVT1 and peptide fragment in tumor growth prediction, metastasis prediction, prognosis evaluation and treatment
  • Application of CircRNA PVT1 and peptide fragment in tumor growth prediction, metastasis prediction, prognosis evaluation and treatment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0145] Embodiment 1: the preparation of a kind of detection kit

[0146] Reagent test kit:

[0147] In this embodiment, a detection kit (including reagent a, reagent b, reagent c, and reagent d) was prepared according to the following composition. The kit is suitable for detecting the expression of PVT1-104aa in biological samples by immunohistochemical methods. The kit may also include a container containing the above reagents and instructions for use.

[0148] Reagent a: blocking solution, which is 10% goat serum (purchased from Suleibao);

[0149] Reagent b: rabbit anti-human or mouse PVT1-104aa polyclonal antibody, antibody name PV-24aa, sequence CSGAQLGLRPDLLAR;

[0150] Reagent c: HRP-labeled goat anti-rabbit secondary antibody (purchased from Suleibao);

[0151] Reagent d: DAB substrate buffer and chromogenic solution (purchased from Solebol).

[0152] Detection method:

[0153] The steps of using the detection kit in this embodiment for detection are as follows: ...

Embodiment 2

[0166] Embodiment 2: the preparation of another kind of detection kit

[0167] Reagent test kit:

[0168] A detection kit (including reagent a, reagent b, reagent c, and reagent d) is prepared according to the following composition, and the kit is suitable for detecting the expression of PVT1-104aa in biological samples by Western blot method. The kit may also include a container containing the above reagents and instructions for use.

[0169] Reagent a: blocking solution, which is 5% skimmed milk powder (purchased from Suleibao Company);

[0170] Reagent b: rabbit anti-human or mouse PVT1-104aa polyclonal antibody (customized), antibody name PV-24aa, sequence CSGAQLGLRPDLLAR;

[0171] Reagent c: HRP-labeled goat anti-rabbit secondary antibody (purchased from Suleibao);

[0172] Reagent d: ECL chemiluminescence detection kit (purchased from Solebol).

[0173] Detection method:

[0174] The steps of using the detection kit in this embodiment for detection are as follows:...

Embodiment 3

[0201] Example 3: Using RT-qPCR to detect the expression of CircRNA PVT1 in human tumor tissues

[0202] Human tumor samples (breast cancer, lung cancer, pancreatic cancer, colon cancer) were selected, and the controls were paracancerous tissues of the respective tumors. TRIzol (Invitrogen, USA) was used to extract tissue total RNA, and qRT-PCR was detected on ABI StepOnePlus TM PCR system (Life Technologies) real-time quantitative PCR instrument to complete.

[0203] The primers and product sizes used are shown in the table below:

[0204]

[0205] CircRNA PVT1 (hsa_circ_0001821) relative quantification using 2- ΔΔCt Calculation by the method (GAPDH is the internal reference) refer to: Livak, KJ. ​​et al., Analysis of relative gene expression data using real-timequantitative PCR and the 2 -ΔΔCt method. Methods. 2001;25:402-408.

[0206] The result is as Figure 12 shown, from Figure 12 The results of RT-qPCR analysis found that CircRNA PVT1 was highly expressed i...

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Abstract

The invention belongs to the field of biotechnology and medicine and specifically relates to correlation and application of a CircRNA PVT1 (hsa _ circ_ 0001821) nucleic acid fragment and a CircRNA PVT1 translated peptide fragment PVT1-104aa in tumor growth prediction, metastasis prediction, prognosis evaluation and treatment. The CircRNAPVT1 and PVT1-104aa are used as drug targets for early diagnosis of tumors, treatment of tumors, alleviation or prevention of tumor metastasis and / or improvement of tumor prognosis. The tissue specificity and stability of CircRNA provided by the invention enable CircRNA to become a good biomarker, and tests prove that the CircRNA PVT1 nucleic acid fragment and the peptide fragment PVT1-104aa play a significant and stable role in tumor growth prediction, metastasis prediction, prognosis evaluation and treatment.

Description

technical field [0001] The present invention belongs to the fields of biotechnology and medicine, and specifically relates to the correlation and application of CircRNA PVT1 (hsa_circ_0001821) nucleic acid fragment and the peptide segment PVT1-104aa translated by circRNA PVT1 in tumor growth prediction, metastasis prediction, prognosis assessment and treatment. Background technique [0002] CircRNA (circular RNA) can be used as a potential biomarker molecule for medicine or clinical diagnosis. Studies have shown that in addition to linear RNA molecules, there are also a large number of closed circular RNA molecules in living organisms, which are specifically expressed in individual development, different tissues and diseases. It was identified by sequencing that these circRNA molecules are usually formed from the 2-4 exons of protein-coding genes through the complementary combination of the 5' end of the front exon and the 3' end of the following exon. Due to the circular s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886G01N33/574A61K45/00A61P35/00
CPCC12Q1/6886G01N33/57484A61K45/00A61P35/00C12Q2600/118C12Q2600/136C12Q2600/158C12Q2600/178G01N2800/52
Inventor 陈地灵郭殷锐吴清平谢意珍祁龙凯唐晓萃杨柏华
Owner GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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