Long-chain non-coding RNA-APOC1P1-3 gene and application thereof for preparing target markers

A non-coding and gene technology, applied to the long-chain non-coding RNA-APOC1P1-3 gene and its use in the preparation of target markers, can solve the problems such as no regulatory effect yet, and improve the level of diagnosis and treatment Effect

Active Publication Date: 2016-12-14
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Existing studies have found that long non-coding RNA (Long non-coding RNA, lncRNA) is closely related to the occurrence and development of various major human diseases, but so far there has been no related lncRNA and its regulatory role in breast cancer, as well as LncRNA-APOC1P1-3 The role of , especially its relationship with tubulin has been reported

Method used

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  • Long-chain non-coding RNA-APOC1P1-3 gene and application thereof for preparing target markers
  • Long-chain non-coding RNA-APOC1P1-3 gene and application thereof for preparing target markers
  • Long-chain non-coding RNA-APOC1P1-3 gene and application thereof for preparing target markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Expression profile of differential lncRNA in breast cancer tissue

[0042] Five isolated fresh human breast cancer tissue specimens and paired paracancerous tissue specimens (from the Department of Breast Surgery, Cancer Hospital Affiliated to Fudan University, with informed consent from the patient) were used for microarray analysis; the following steps were followed:

[0043] (1) Use TRIzol reagent to extract total RNA from tissue samples, and use RNasey Mini Kit to purify the extracted RNA;

[0044] (2) Use QuickAmp Labeling Kit, One-Color (Agilent p / n 5190-0442) to synthesize and label double-stranded cDNA;

[0045] (3) After the labeled double-stranded cDNA is purified, it is hybridized to the Human 8x60K LncRNA expression array information chip of Arraystar;

[0046] (4) After hybridization and rinsing, scan and analyze by Agilent Microarray Scanner (Agilent p / n G2565BA) scanner;

[0047] (5) The original data analysis was completed by Agilent Feature ...

Embodiment 2

[0048] Example 2, RNAPull Down

[0049] (1) Plasmid digestion

[0050] Reaction ingredients and conditions:

[0051]

[0052] (2) Linear DNA identification

[0053] Use ordinary agarose gel electrophoresis (TBE preparation) to detect whether the plasmid DNA is cut into linearity, 1 μl DNA and 5 μl 6x sample buffer are thoroughly mixed, add 1.5% agarose gel to the sample hole, electrophoresis at 60V for 25min, and observe the bands by ultraviolet light , Tianneng gel imaging system takes photos and analyzes;

[0054] (3) Linear DNA purification

[0055] 1) Add an equal volume of Solution I to the enzyme digestion reaction and mix well (if the DNA sample is 20 μl, add 20 μl of Solution I); 2) Add it to the DNA purification column and place it at room temperature for 1 min; 3) Centrifuge at 12000rmp for 1 min and discard Collect the liquid in the tube; 4) Add 700 μl solution II to the DNA purification column, and let it stand at room temperature for 1 min; 5) Centrifuge at...

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Abstract

The invention relates to the field of bio-medicines and relates to a long-chain non-coding RNA expression profile and an APOC1P1-3 gene of breast cancer, and the application thereof for preparing target markers. A test proves that the APOC1P1-3 gene can be combined with tubulin protein, can inhibit breast cancer cell apoptosis and improve breast cancer cell proliferation and further has effects on generation and development of the breast cancer. The invention also provides a new target for diagnosis and therapy of the breast cancer. The long-chain non-coding RNA-APOC1P1-3 gene is used for preparing the target markers, and especially, can be used for preparing the new target for the diagnosis and therapy of the breast cancer. The invention has important clinical significance on improvement of diagnosis level of the breast cancer.

Description

technical field [0001] The present invention relates to the field of biomedicine, in particular to breast cancer long-chain non-coding RNA expression profile and APOC1P1-3 gene, in particular to long-chain non-coding RNA-APOC1P1-3 gene and its use in preparing target markers, Especially the long-chain non-coding RNA-APOC1P1-3 gene and its use in preparing new target markers for diagnosis and treatment of breast cancer. Background technique [0002] Breast cancer is one of the most common malignant tumors in Chinese women, and has become the number one killer that seriously threatens the health and life of Chinese women in recent years. Studies have shown that postoperative breast cancer is prone to metastasis, which is closely related to the growth, invasion and metastasis of breast cancer cells to a large extent. Therefore, professionals in the industry believe that the study of molecular markers and gene therapy targets for breast cancer diagnosis is of great importance I...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 周平廖晓红吕雷于建刚杨佳音陆齐李录英
Owner FUDAN UNIV
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