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LRPPRC (leucine rich pentatricopeptide repeat containing) specific nucleic acid aptamer and application thereof

A nucleic acid aptamer and nucleic acid aptamer technology, applied in the field of medicine, can solve the problems of unclear LRPPRC target effect and treatment mechanism, and achieve the effects of enhanced killing effect, enhanced therapeutic effect, and good tissue permeability

Active Publication Date: 2018-08-17
INST OF CHEM CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether LRPPRC can be used as a target for tumor treatment, the effect of tumor treatment and the mechanism of treatment are still unclear.

Method used

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  • LRPPRC (leucine rich pentatricopeptide repeat containing) specific nucleic acid aptamer and application thereof
  • LRPPRC (leucine rich pentatricopeptide repeat containing) specific nucleic acid aptamer and application thereof
  • LRPPRC (leucine rich pentatricopeptide repeat containing) specific nucleic acid aptamer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1, the synthesis of nucleic acid aptamer and its pretreatment

[0042]Nucleic acid aptamer R14 without any modification: 5'-GGTGGGTGGGTTGGGTGG-3' (SEQ ID NO: 1, wherein G and T represent unmodified guanine and thymine, respectively). Synthesized by AB 3400 DNA synthesizer, the synthesized DNA was purified by HPLC, vacuum-dried, 80% acetic acid to remove DMT, ethanol precipitation and vacuum-dried, adjusted to pH 7.0 with Tris-HCl (pH 9.0), filtered and sterilized, and finally used without Bacteria were diluted to 500 μM in D-PBS and stored at -20°C for later use.

Embodiment 2

[0043] Example 2, the growth of nucleic acid aptamer R14 on A549 and A973 cells in vitro

[0044] After A549 cell-grade A973 cells were treated with trypsin-EDTA and centrifuged, they were planted in a 96-well plate of xCELLigence RTCA MP system label-free cell function analyzer at a density of 2000 cells / well and cultured for 24 hours. The fresh medium containing LRPPRC-specific nucleic acid aptamer R14 was replaced with a concentration gradient of 5 μM, 10 μM, 20 μM, 40 μM, or negative control sequence RTT (40 μM), and cultured for 24 h. For each treatment, 6 groups of parallel experiments were set up. Cultivate and test, draw cell growth rate curve, see results figure 1 . Experiments show that the aptamer R14 can significantly inhibit the growth of lung cancer A549 and A973 in vitro, and the effect is concentration-gradient dependent.

Embodiment 3

[0045] Example 3. Influence of nucleic acid aptamer R14 combined with SRC kinase inhibitor on the proliferation of A549 and A973 cells in vitro

[0046] After A549 cells and A973 cells were treated with trypsin-EDTA and centrifuged, they were planted in a 96-well plate of xCELLigence RTCA MP system label-free cell function analyzer at a density of 2000 cells / well and cultured for 24 hours. Replace with fresh medium containing LRPPRC-specific nucleic acid aptamer R14 or an irrelevant control sequence at a concentration of 5 μM, and culture for 24 hours. Add the corresponding SRC kinase inhibitor, continue to culture and detect, set 6 groups of parallel experiments for each group of treatments, see the results figure 2 . Experiments have shown that 5 μM aptamer R14 can significantly increase the sensitivity of lung cancer A549 and A973 to SRC kinase in vitro, and the combination of the two can effectively kill tumor cells.

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Abstract

The invention relates to a nucleic acid aptamer R14 which can specifically recognizing LRPPRC (leucine rich pentatricopeptide repeat containing), and the sequence of the nucleic acid aptamer R14 is shown in SEQ ID NO: 1. The nucleic acid aptamer R14 is capable of inhibiting tumor cell proliferation by blocking the normal tissue function of the LRPPRC and interfering the cell cycle progression. Theinvention further provides application of the nucleic acid aptamer R14 specifically recognizing the LRPPRC and SRC kinase inhibitor in combined treatment of tumour. The nucleic acid aptamer R14 is capable of enhancing the sensitivity of tumour to the SRC kinase inhibitor, and the SRC kinase inhibitor can be used for eliminating SRC kinase activity abnormal activation caused by the nucleic acid aptamer R14.

Description

technical field [0001] The invention belongs to the field of medicine, and relates to a nucleic acid aptamer drug for treating tumors, in particular to a nucleic acid aptamer drug specifically recognizing LRPPRC and its use in treating tumors, more specifically LRPPRC-specific nucleic acid aptamers and SRC kinase inhibitors application of agents in the treatment of tumors. Background technique [0002] Cancer has developed into the number one killer of human health, and the number of deaths from cancer worldwide is about 7 million every year. Chemotherapy of cancer has been well developed with the development of cell biology and the understanding of cancer pathogenesis. At present, there are about 90 kinds of chemical drugs developed for killing tumor cells and treating cancer. However, these drugs lack specificity and often bring fatal side effects to patients in clinical practice. Therefore, it is necessary to develop targeted drugs that specifically kill tumors or inhi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/115A61K31/7088A61K45/06A61P35/00
CPCA61K45/06A61P35/00A61K31/7088C12N15/115C12N2310/315C12N2310/3183C12N2310/3181C12N2310/16
Inventor 方晓红周卫徐丽赵立波
Owner INST OF CHEM CHINESE ACAD OF SCI
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