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Long non-coding RNA (Ribonucleic Acid) and application thereof

A non-coding, non-small cell lung cancer technology, applied in the application field of preparing drugs for the treatment of non-small cell lung cancer, can solve the problems of lost surgical opportunities, low survival rate, poor chemotherapy regimen for lung cancer, etc.

Inactive Publication Date: 2015-12-23
THE SECOND AFFILIATED HOSPITAL OF NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many lung cancer patients have metastasized at the time of diagnosis, and often lose the chance of surgery, and the effect of chemotherapy for lung cancer is not good, and the five-year survival rate is less than 5%. It is urgent to find new and effective methods for the treatment of lung cancer

Method used

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  • Long non-coding RNA (Ribonucleic Acid) and application thereof
  • Long non-coding RNA (Ribonucleic Acid) and application thereof
  • Long non-coding RNA (Ribonucleic Acid) and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Detection of the expression of lnc-uc003hsl.1 in tissues and cells

[0055] (1): The expression of lnc-uc003hsl.1 in normal lung tissue and lung cancer tissue:

[0056] 1. Chip preparation and analysis: Prepare normal lung tissue and lung cancer tissue samples according to the requirements of Boao Company in China, and submit them to Boao Company for chip preparation. The human long-chain non-coding RNA chip V1.0 version completed chip analysis, which contains the latest versions of many LncRNAs databases and tens of thousands of probes for LncRNAs detection. The detection principle of the chip is based on the length and structural characteristics of the lncRNA sequence, using a combination of random primers and Ologo(dT) reverse transcription to introduce the T7 promoter, and then amplifying the RNA by in vitro transcription-mediated linear amplification , RNA linear amplification method for sample labeling in microarray chip analysis. The DNA obtained by reverse tr...

Embodiment 2

[0074] Construct lnc-uc003hsl.1 lentiviral interference vector to transfect cells:

[0075] Design a specific interfering sequence against lnc-uc003hsl.1, its sequence is (5'to3'): CCAGATGGGCACTTCTAAA, see SeqNO.2, take the sequence fragment interfering with the GAPDH gene as a control, and insert it into the lentiviral vector ( The above vectors and fragments were synthesized by Invitrogen Company). The packaged lentiviral vector was used to infect the non-small cell lung cancer cell line A549 to down-regulate the expression of lnc-uc003hsl.1. After 48 hours, the stably transfected cell line was screened with G418 and named A549 / sh-lnc-uc003hsl. 1 (control is A549 / lnc-NC).

Embodiment 3

[0077] MTT method to detect cell proliferation:

[0078] 1) Cell inoculation: Digest monolayer cultured cells with 0.25% trypsin, prepare a single A549 / sh-lnc-uc003hsl.1 (control is A549 / lnc-NC) cell suspension with 1640 culture medium containing 10% fetal bovine serum 2500 cells per well were inoculated in a 96-well culture plate with a volume of 200ul per well. DDP (cisplatin, purchased from Jiangsu Hengrui Medicine Co., Ltd.) or DTX (docetaxel, purchased from Jiangsu Hengrui Medicine Co., Ltd.) were administered respectively.

[0079] 2) Culture cells: move the culture plate into CO 2In an incubator at 37°C, 5% CO 2 and relative humidity conditions for 3 days.

[0080] 3) Color development: After 6h, 24h, 48h, 72h, and 96h of culture, add 20ul of MTT solution (5mg / ml) to each well, continue to incubate at 37°C for 4h, terminate the culture, and carefully discard the culture supernatant in the well. Add 150 μl DMSO (dimethyl sulfoxide) and shake for 10 min to fully disso...

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Abstract

The invention belongs to the field of genetic engineering and particularly relates to application of long non-coding RNA (Ribonucleic Acid) uc003hsl.1 in preparation of drugs for treating non-small cell lung cancer. The influence on the apoptosis, propagation, drug susceptibility and the like of non-small cell lung cancer cells is caused through changing the expression of the long non-coding RNA uc003hsl.1, so that through lowering the expression of the long non-coding RNA uc003hsl.1, the apoptosis of the non-small cell lung cancer cells can be promoted, the propagation of the non-small cell lung cancer cells can be inhibited, and the susceptibility to chemotherapy drugs of the non-small cell lung cancer cells can be enhanced.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and particularly relates to the application of long non-coding RNA uc003hsl.1 in the preparation of drugs for treating non-small cell lung cancer. Background technique [0002] Genomic studies have shown that there are about 20,000 protein-coding genes in the human genome, accounting for about 2% of the total number of genes, and most of the remaining genes are transcribed into non-coding RNA (non-coding RNA, ncRNA). According to their length, ncRNAs can be divided into transcription initiation RNAs, Piwi protein-interacting RNAs, microRNAs, small nucleolar RNAs, and long non-coding RNAs (longnon-coding RNAs, lncRNAs). LncRNA is a class of RNA molecules whose transcripts are more than 200 nucleotides in length and do not encode proteins themselves. LncRNA was originally considered as a by-product of RNA polymerase II transcription, which is the "noise" of genome transcription and has no biolog...

Claims

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Application Information

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IPC IPC(8): C12N15/113A61K48/00A61P35/00
Inventor 王朝霞魏晨晨陈沁楠姜黎黎
Owner THE SECOND AFFILIATED HOSPITAL OF NANJING MEDICAL UNIV
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