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Application of long-chain non-coding RNA in preparation of non-small cell lung cancer treatment drugs

A non-small cell lung cancer, long-chain non-coding technology, applied in the field of genetic engineering, can solve problems such as low survival rate, lost chance of surgery, poor chemotherapy effect of lung cancer, etc.

Active Publication Date: 2013-09-25
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 opportunity for surgery, and the chemotherapy regimen for lung cancer is not effective, and the five-year survival rate is less than 5%. There is an urgent need to find new and effective methods for the treatment of lung cancer

Method used

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  • Application of long-chain non-coding RNA in preparation of non-small cell lung cancer treatment drugs
  • Application of long-chain non-coding RNA in preparation of non-small cell lung cancer treatment drugs
  • Application of long-chain non-coding RNA in preparation of non-small cell lung cancer treatment drugs

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] The inventor provided samples of normal lung tissue and lung cancer tissue, which were submitted to Boao Company for chip preparation. The R human long-chain non-coding RNA chip V1.0 version completed the chip analysis. The chip contains the latest versions of many LncRNAs databases, and currently covers about 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 Oligo(dT) reverse transcription to introduce the T7 promoter, and then amplifying the RNA by linear amplification mediated by in vitro transcription. RNA linear amplification method is used for sample labeling in microarray chip analysis. The linear amplification product cRNA is reverse transcribed to obtain DNA, and then the DNA is fluorescently labeled with KlenowFragent enzyme, and finally the fluorescently labeled DNA product is hybridized with the Oligo p...

Embodiment 2

[0063] Example 2 Detection of the expression of lnc-uc002llc.1 in tissues and cells

[0064] (1): The expression of lnc-uc002llc.1 in normal lung tissue and lung cancer tissue:

[0065] 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. Jingxin○R human long non-coding RNA chip V1.0 version completed the chip analyze.

[0066] 2. Chip results: The experimental data comes from Boao company, the results are as follows figure 1 And shown in Table 1.

[0067] Table 1 list of significant differences

[0068]

[0069] 3. Analysis of results: The signal of lnc-uc002llc.1 in lung cancer tissue (Sample B) is 339.8561, the signal in normal lung tissue (Sample A) is 9.2367, and the Ratio (Sample B / Sample A) is 36.7941, indicating lnc-uc002llc .1 is highly expressed in lung cancer tissue and low in normal lung tissue, with a differenc...

Embodiment 3

[0076] Embodiment 3MTT method detects cell proliferation:

[0077] 1) Cell inoculation: Digest monolayer cultured cells with 0.25% trypsin, and prepare a single A549 / sh-lnc-uc002llc.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.

[0078] 2) Culture cells: move the culture plate into CO 2 In an incubator at 37°C, 5% CO2 and relative humidity conditions for 3 days.

[0079] 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 t...

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Abstract

The invention belongs to the genetic engineering field, and especially relates to an application of long-chain non-coding RNA in the preparation of non-small cell lung cancer treatment drugs. The change of the lnc-uc002llc.1 expression has influences on the apoptosis, proliferation, the drug susceptibility and the like of non-small cell lung cancer cells, so the reduction of the lnc-uc002llc.1 expression can realize the apoptosis promotion, proliferation inhibition and chemotherapy drug susceptibility enhancement of the non-small cell lung cancer cells.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and particularly relates to the application of lnc-uc002llc.1 in the preparation of medicines for treating non-small cell lung cancer. Background technique [0002] RNA is one of the most important material bases in organisms, and together with DNA and proteins, it constitutes the framework of life. But for a long time, RNA was thought to be just a "transition" between DNA and protein, from which it gets its sequence and then converts the genetic information into protein. However, a series of studies have shown that these small RNAs actually manipulate many cellular functions, and they can turn off or regulate gene expression by reacting against DNA through the binding of complementary sequences. A recently discovered class of non-protein-coding RNAs, long non-coding RNAs (Long non-coding RNAs, lncRNAs) are a class of RNA molecules with transcripts longer than 200 nt, they do not encode protei...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61P35/00
Inventor 王朝霞刘志利刘晶
Owner THE SECOND AFFILIATED HOSPITAL OF NANJING MEDICAL UNIV
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