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Application of long-chain non-coding RNA in preparing preparation for inhibiting tumor cell migration

A long-chain non-coding, tumor cell technology, applied in anti-tumor drugs, medical preparations containing active ingredients, gene therapy, etc.

Active Publication Date: 2018-07-03
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] So far, there are no reports about the role of long non-coding RNA molecules in the development of tumors, especially how

Method used

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  • Application of long-chain non-coding RNA in preparing preparation for inhibiting tumor cell migration
  • Application of long-chain non-coding RNA in preparing preparation for inhibiting tumor cell migration
  • Application of long-chain non-coding RNA in preparing preparation for inhibiting tumor cell migration

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0168] Preparation of competent bacteria:

[0169] (1) Inoculate the Top 10 strain on an LB plate without ampicillin and culture for 8h-12h.

[0170] (2) Pick a single clone to an LB liquid medium without ampicillin, and culture it overnight at 37°C with shaking at 220 r / min.

[0171] (3) Inoculate the above-mentioned culture broth in 200 mL of fresh LB liquid medium at a ratio of 1:100, and cultivate to the logarithmic growth phase (OD value about 0.6) at 37° C. and 220 r / min with shaking.

[0172] (4) Dispense the cultured bacteria liquid into 50 mL centrifuge tubes, and ice bath for 30 minutes. Centrifuge at 2400 rpm for 10 min, and discard the supernatant.

[0173] (5) Add 10mL 0.1mol / L pre-cooled CaCl2 to suspend bacteria. 2400rmp, discard the supernatant.

[0174] (6) Add 10mL of 0.1mol / L pre-cooled CaCl2 to suspend the bacteria, place on ice for 30min, 2400rpm, discard the supernatant.

[0175] (7) Add 1 mL of 0.1 mol / L pre-cooled CaCl2 to suspend the bacteria, divide them into 1...

Example Embodiment

[0331] Example 1 Long non-coding RNA BC009639 inhibits lung cancer cell migration

[0332] 1.1 Lung cancer cell long non-coding RNA expression chip data

[0333] A pair of lung cancer cell lines 95C and 95D with different metastatic potentials were selected as the comparative study objects. Among them, the metastatic ability of 95D was higher than that of 95C. The long non-coding RNA chip was used to obtain the long non-coding RNA expression of each of the pair of cell lines. Data, in the obtained data, by means of bioinformatics analysis, screening the relatively high expression abundance in this pair of cells, and a high degree of expression difference long-chain non-coding RNA, of which the long-chain non-coding RNA BC009639 has a relatively high expression level, and more importantly, its expression level difference between 95C and 95D is very significant, and it is selected as a further research object;

[0334] 1.2 RT-PCR verifies that BC009639 is expressed in 95C and D

[0335...

Example Embodiment

[0346] Example 2 Function experiment of LncRNA BC009639 affecting MiR-96 and MiR-155

[0347] lncRNA BC009639 sequence analysis

[0348] Analyzing the sequence of lncRNA BC009639, BLAST results showed that in the human gene database, lncRNA BC009639 highly matched an mRNA sequence named IMPAD1. The complete sequence of lncRNA BC009639 showed the characteristics of overlap with part of the IMPAD1 mRNA sequence, and their matching regions Mainly in the 3'UTR region of IMPAD1 mRNA sequence;

[0349] Correlation between IMPAD1 and lncRNA BC009639

[0350] Based on the important role of IMPAD1 gene for physiological development, further study its expression level in various tumor cells, and detect its expression level in lung cancer, liver cancer, colorectal cancer and other different tumor cells by real-time PCR. The results show that it has a relatively high expression level in various types of tumor cells. Based on the correlation between IMPAD1 and lncRNABC009639 in the base sequence,...

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Abstract

The invention belongs to the technical field of gene, and relates to an application of long-chain non-coding RNA BC009639 in preparing a preparation for inhibiting tumor cell migration. In the application, a long-chain non-coding RNA BC0009639 possibly closely related to tumor migration is screened and verified from a pair of lung cancer cell lines with different migration potency by using a genechip, proved by scratch experiment and Transwell experiment, the long-chain non-coding RNA BC0009639 can inhibit migration of tumor cells, and proved by MTT experiment, the long-chain non-coding RNA BC0009639 can inhibit the multiplication capacity of the cells; the experiment results verify that the long-chain non-coding RNA BC009639 can inhibit the migration of tumor cells and inhibit the multiplication capacity of the cells; the sequence of the long-chain non-coding RNA BC0009639 is closely related to 3'UTR of an IMPAD1 mRNA sequence, and the expression is affected by gambogic acid and is in negative correlation with the survival rate of tumor cells. The invention provides basic theory foundation and research data for exploring the action mechanism of the long-chain non-coding RNA BC0009639 to an organism and correlation and directed treatment of clinical diseases.

Description

technical field [0001] The invention belongs to the field of gene technology, and specifically relates to long-chain non-coding RNA BC009639 and its use in the preparation of preparations for inhibiting tumor cell metastasis. The long-chain non-coding RNA BC009639 can inhibit tumor cell migration and cell proliferation. It can inhibit the metastasis of tumor cells; the sequence of the long-chain non-coding RNA BC0009639 is closely related to the 3'UTR of the IMPAD1 mRNA sequence, its expression is affected by gambogic acid, and it is negatively correlated with the survival rate of tumor cells. Background technique [0002] It is reported in the prior art that a tumor is a tissue formed by abnormal proliferation and differentiation of local tissue cells in the human body after losing their normal ability to regulate their growth. Once a tumor is formed in the body, its proliferation is not subject to the physiological regulation of the body and destroys the physiological func...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K31/7105A61P35/00A61P35/04
CPCA61K31/7105
Inventor 吴兴中陈忠义
Owner FUDAN UNIV
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