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PICT-1 protein truncated mutant and application thereof

A technology of PICT-1 and mutants, applied in PICT-1 protein truncated mutants and its application fields, can solve the problems of tumor suppressor function and molecular mechanism that are not clear

Inactive Publication Date: 2015-12-23
SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, its tumor suppressor function and molecular mechanism are still far from clear.

Method used

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  • PICT-1 protein truncated mutant and application thereof
  • PICT-1 protein truncated mutant and application thereof
  • PICT-1 protein truncated mutant and application thereof

Examples

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Embodiment 1

[0033] GLTSCR2 is considered a tumor suppressor gene. In 2004, Okahara et al. used yeast two-hybrid technology to screen the human brain cDNA library to obtain a positive clone using the C-terminus of PTEN as a bait protein. They named it PICT-1 and found that its coding gene was GLTSCR2. Subsequent studies have shown that PICT-1 can interact with the important tumor suppressor gene PTEN, promote the phosphorylation and stability of PTEN in the cytoplasm, and then inhibit the occurrence and growth of tumors. However, recent studies have found that PICT-1 is a nucleolus-localized protein.

[0034] Based on the above, the inventors believe that in addition to the PTEN-dependent anticancer effect in the cytoplasm, it probably also plays an important role in the nucleolus.

[0035] This example shows the inventors' experimental process based on the above knowledge and assumptions, design and verification of the following findings: overexpression of PICT-1 can induce autophagy in ...

Embodiment 2

[0061]As described in Example 1, the ability of PICT-1 protein to induce autophagy is related to its nucleolar localization. The nucleolus is the most prominent structure in the interphase nucleus of eukaryotic cells. The nucleolus consists of three major parts: the fibrous center (FC), the dense fibrous component (DFC), and the granular component (GC). The nucleolus is the site of rDNA gene storage, rRNA synthesis and processing, and ribosomal subunit assembly. Ribosome biogenesis includes RNA transcription, processing and assembly of ribosomal subunits, which is a vector process. Transcription mainly occurs in the FC region, then rRNA is processed and modified in the DFC region, and assembled into ribosome-sized subunits in the GC region. In order to study whether PICT-1 protein can regulate the transcription of rDNA and the process of ribosome, and then affect the occurrence of autophagy. The inventors investigated the subcellular localization of PICT-1 in the nucleolus,...

Embodiment 3

[0092] It has been reported that the inhibition of AKT / mTOR / p70S6K cell signaling is closely related to the occurrence of autophagy. To investigate whether autophagy induced by PICT-1 overexpression is related to AKT / mTOR / p70S6K cell signaling. We investigated the phosphorylation levels of AKT, mTOR and p70S6K proteins. Proceed as follows:

[0093] The day before, with 1.0 x 10 6 Seed cells at a density of 6 wells.

[0094] 2. On the second day, according to the transfection method in step 2 in Example 1, pFLAG-CMV2, pFLAG-CMV2-PICT-1, pFLAG-CMV2-PICT-1(1-346), pFLAG-CMV2-PICT- 1(181-346) or pFLAG-CMV2-PICT-1(181-479) were transfected into U251 cells.

[0095] 3. Collect the cells at an appropriate time point after transfection, and use β-actin as an internal reference for Western blot detection.

[0096] The result is as Figure 4 Shown: the result is as Figure 4 As shown in A and 4B, overexpression of PICT-1 inhibited the phosphorylation levels of AKT, mTOR and p70S6...

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Abstract

The invention discloses a PICT-1 (protein interacting with carboxyl terminus 1) protein truncated mutant which has a sequence represented by sites from 181 to 478 in SEQ ID NO: 1. Due to the overexpression of the truncated mutant, the UBF activity can be adjusted, rDNA transcription can be suppressed, and cell autophage can be caused; the PICT-1 protein truncated mutant can be used for preparing an anti-tumor medicament, particularly a medicament for treating glioma. The invention further discloses a nucleic acid for encoding the truncated mutant and application of the nucleic acid, an expression vector of the nucleic acid for encoding the truncated mutant, a host cell comprising the expression vector, and a method and a device for treating the glioma.

Description

technical field [0001] The present invention relates to the field of biomedicine, specifically, the present invention relates to PICT-1 protein truncated mutants and applications thereof, more specifically, the present invention relates to a PICT-1 protein truncated mutant, PICT-1 protein truncated mutant Use of body, truncated mutant encoding nucleic acid, use of truncated mutant encoded nucleic acid, expression vector of truncated mutant encoded nucleic acid, a host cell comprising the expression vector, a method and device for treating glioma . Background technique [0002] Glioma originates from glial cells in the brain and is the most common primary brain tumor in adults, especially young adults, accounting for 40-50% of primary intracranial tumors. [0003] In human glioma, a 150kb fragment (19q13.32) on the long arm of chromosome 19 often has an allelic deletion. This shows that there may be tumor suppressor genes on this chromosome, and the deletion of these genes ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C12N15/12C12N15/85C12N15/66C12N5/10A61K38/17A61P35/00
CPCA61K38/00C07K14/47
Inventor 黄来强陈红波
Owner SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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