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Radiotherapy-targeted sensitizer and application thereof

A technology of radiotherapy and sensitizers, which is applied in gene therapy, antineoplastic drugs, DNA/RNA fragments, etc., can solve the problems of tumor cell radiotherapy resistance, trouble radiotherapy effect, and no reports, so as to improve radiotherapy sensitivity and radiotherapy Effect of increased sensitivity, good economic and social benefits

Active Publication Date: 2014-11-12
HANGZHOU NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the radioresistance of tumor cells is still a problem that seriously plagues the radiotherapy effect.
At present, the study on the regulation of miRNAs related to radiotherapy resistance in laryngeal cancer cells, especially laryngeal cancer stem cells, has not been reported at home and abroad.

Method used

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  • Radiotherapy-targeted sensitizer and application thereof
  • Radiotherapy-targeted sensitizer and application thereof
  • Radiotherapy-targeted sensitizer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Screening of radiotherapy difference miRNAs

[0035] 1.1 Culture of Hep-2 stem cells

[0036] Prepare serum-free medium (SFM): add 5 μl each of EGF and bFGF to 5 ml of DMEM / F12 culture medium at a volume ratio of 1000:1.

[0037] Human laryngeal carcinoma epithelial cells Hep-2 in the logarithmic growth phase were resuspended in SFM medium to make 10 cells 3 cell suspension per ml, at 25cm 2 Routine culture in cell culture flasks, i.e. vertically place the culture flasks in an environment containing 5% CO 2 In a cell culture incubator at 37°C, shake several times a day. Observe the formation of microspheres under a microscope. In the ultra-clean bench, change half of the medium every 2 days, that is, discard half of the SFM medium, add an equal amount of fresh SFM medium, and pass on every 6 days. The number of cells is 1:3. Add the culture medium in a 1.5ml centrifuge tube, collect the microspheres by centrifugation at 4°C, 1000g, 4 minutes, digest with t...

Embodiment 2

[0062] Example 2Hep-2 cell transfection and transfection rate determination

[0063] 2.1 Transfection of miRNA in Hep-2 cells

[0064] 2.1.1 Radiotherapy targeted sensitization group preparation: Taking 24-well cell culture plate as an example, (1) the 7 kinds of oligonucleotides synthesized in Example 1 were prepared in a mass ratio of 1:1:1:1:1:1 :1 After mixing, samples with a total mass of 0.33 μg, 0.67 μg, and 1 μg were prepared, dissolved in 2.1 μl, 4.2 μl, and 6.3 μl of DEPC water in turn, and then 22.9 μl, 20.8 μl, and 18.7 μl of RPMI1640 culture solution were added , make a final volume of 25 μl RNA dilution, and let it stand at room temperature for 5 minutes; (2) Mix 2 μl of Entranstr-R with 23 μl of RPMI1640 culture medium to make a final volume of 25 μl of Entranstr-R dilution, and let it stand at room temperature Set aside for 5 minutes; mix equal volumes of the RNA diluent prepared in (1) and (2) and the Entranstr-R diluent to make 50 μl sensitizer transfection ...

Embodiment 3

[0074] Example 3 Analysis of biological characteristics of Hep-2 cells after radiotherapy

[0075] 3.1 Radiotherapy of Hep-2 cells after transfection

[0076] With 6 groups of Hep-2 cells in Example 2 (group 1 is transfection 50nM sensitizer group, group 2 is transfection 100nM sensitizer group, group 3 is transfection 150nM sensitizer group, group 4 is transfection The nonsense miNA group, group 5 was the transfection PBS group, and group 6 was the non-transfection blank group) received radiotherapy immediately, the conditions were 6MV-X-ray vertical irradiation, the total dose was 400cGy, the dose rate was 200cGy / min, and the single dose The irradiation field was 200cGy, and the irradiation field was 20cm×20cm. During irradiation, the surface of the culture plate was covered with 1.5cm tissue-equivalent filler, and the source-to-skin distance (SSD) was 100cm. Irradiate once at intervals of 48 hours, and irradiate twice in total.

[0077] 3.2 Hep-2 cell proliferation experi...

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Abstract

The invention discloses a radiotherapy-targeted sensitizer, comprising the following seven miRNAs: an hsa-miR-138-2-3p stimulant, an hsa-miR-208b-3p stimulant, an hsa-miR-197-3p inhibitor, an hsa-miR-20b-3p inhibitor, an hsa-let-7a-5p inhibitor, an hsa-miR-16-1-3p inhibitor and an hsa-miR-21-5p inhibitor. The invention provides a novel radiotherapy-targeted sensitizer, which is capable of enhancing the radiosensitivity and is small in toxic and side effects, is especially applied to laryngeal cancer patients at various stages, and has an important application prospect. The novel radiotherapy-targeted sensitizer disclosed by the invention is expected to play a great role in improvement of the radiosensitivity and further increase of the curative effect, and good economic and social benefits are generated.

Description

(1) Technical field [0001] The present invention relates to a radiation therapy target sensitizer, in particular to a miRNAs composition and application thereof with a radiation therapy target sensitization effect. (2) Background technology [0002] Cancer stem cells are one of the few cells in tumors with self-renewal, unlimited proliferation and multi-directional differentiation potential, and inherent radiation resistance. It is generally believed that cancer stem cells are the root of tumor development and radiation resistance. In recent years, people have successfully isolated and identified nearly ten kinds of tumor stem cells; the more recognized "marker" cells are side population cells (tumor stem cell-like cells) and CD133 cells detected by flow cytometry. + CD44 + tumor cells. [0003] At present, some studies have gone deep into the regulation of key genes of tumor stem cells. One of the most eye-catching is the study of microRNA (microRNA, miRNAs) in tumor ste...

Claims

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Application Information

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IPC IPC(8): C12N15/113A61K48/00A61P35/00
Inventor 黄常新朱影
Owner HANGZHOU NORMAL UNIVERSITY
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