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Tumor radiation-sensitizing and radiation bystander effect reducing function of miRNA, implementation method, and uses

A tumor radiation and application technology, applied in the fields of biotechnology and medicine, can solve the problems that there are no research reports on miR214

Active Publication Date: 2016-06-01
ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition to the above studies, there is no research report on the role of miR214 and miR495 in tumor radiosensitization and attenuation of radiation bystander damage.

Method used

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  • Tumor radiation-sensitizing and radiation bystander effect reducing function of miRNA, implementation method, and uses
  • Tumor radiation-sensitizing and radiation bystander effect reducing function of miRNA, implementation method, and uses
  • Tumor radiation-sensitizing and radiation bystander effect reducing function of miRNA, implementation method, and uses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Construction and identification of HepG2 stable cell lines overexpressing miR214 and miR495

[0045] Stable cell lines overexpressing miR214 and miR495 were constructed using a lentiviral packaging system (purchased from SBI, USA). The components of the packaged virus were transfected into 293T cells with MegaTran1.0 transfection reagent, and the virus supernatant containing miR214 or miR495 precursor sequence (SEQ ID NO: 1 or SEQ ID NO: 2) was prepared, and then HepG2 cells were infected with the virus supernatant to obtain Stable cell lines incorporating miR214 or miR495 precursor sequences. These stable cell lines can utilize host cells to transcribe RNA precursors and cleavage to mature miR214 or miR495, so they can continuously overexpress miR214 or miR495. At the same time, an empty vector (control) was packaged as a cell line control. Stable cell lines use TRIzol (Invitrogen Company) to extract total cellular RNA, use SYBRQPCR reagent (Exiqon Company)...

Embodiment 2

[0046] Example 2 Verification of the radiosensitizing effect of miR214 and miR495 in vivo and in vitro

[0047] 2.1 In vitro radiation sensitization experiment of miR214 and miR495

[0048] HepG2 stable cells overexpressing miRNA214, miR495, and control group were used for colony formation experiments. The three types of cells were seeded in six-well plates with the same number, and each group was duplicated. After the cells were completely adhered to the wall, 60 The cells were irradiated with Co-γ rays once (dose 6Gy, dose rate 130cGy / min), then the plate was returned to the incubator for incubation, and the medium was replaced every two days. After the cells in the dish grow into cell groups, take out the plate for fixation and staining, and finally take pictures for counting. The results of clone formation experiments showed that the colony formation numbers of overexpressed miR214 group and miR495 group were significantly lower than those of the control group at the same...

Embodiment 3

[0051] Embodiment 3miR495 weakens the in vivo experiment of radiation bystander effect

[0052]At the end of the miR495 in vivo radiation sensitization experiment, the animals in the miR495 group were active and in good condition, while the animals in the control group were thin, sluggish and in poor condition. The two groups of mice were dissected to observe the morphological changes of the organs combined with HE staining. The results showed that multiple necrotic lesions were observed in the liver of the control group, and HE staining supported this phenomenon. are close to normal liver. In vivo experiments showed that overexpression of miR495 could effectively attenuate the liver damage caused by radiation side effects.

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Abstract

The invention belongs to the technical field of biomedicine, and particularly relates to a radiation-sensitizing function of miR214, a radiation-sensitizing and radiation bystander effect damage reducing function of miR495, uses of the miR214 and the miR495 in improvement of a tumor radiotherapy treatment effect, and an application method. In the invention, miRNA can be used as an auxiliary medicine for improving a tumor clinical radiotherapy effect, particularly, the miR495 can improve the tumor radiation sensitivity and can reduce the damage to a liver caused by a bystander effect, and a new idea is provided for preparation of a clinical tumor radiotherapy auxiliary medicine.

Description

technical field [0001] The invention belongs to the field of biotechnology and medicine, and specifically relates to a radiation-sensitizing effect of a small RNA molecule miR214, and a radiation-sensitizing effect of another RNA small molecule miR495, and the effect of reducing radiation side effects, as well as their implementation methods and use. Background technique [0002] MicroRNA (micoRNA, referred to as miRNA) is a kind of non-coding single-stranded RNA with a length of 18-24 nucleotides encoded by an endogenous gene. The miRNA precursor (pre-miRNA) is cleaved by Dicer enzyme (a double-stranded RNA-specific RNA endonuclease). They are involved in post-transcriptional gene expression regulation in animals and plants, and can degrade the target mRNA at the post-transcriptional level by binding to the 3'-UTR region of the target mRNA almost completely, or inhibit protein synthesis at the translational level by combining with it incompletely. The earliest report abou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K41/00A61K31/7105A61P35/00
CPCA61K31/7105A61K41/0038
Inventor 宋海峰叶棋浓付洁张蒙高新王瑜
Owner ACADEMY OF MILITARY MEDICAL SCI