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Application of Smurf1 gene, expression product and derivative or inhibitor thereof in chemotherapy of colorectal cancer

A technology of expression product and gene expression, applied in the field of medicine, can solve the problem of patients' insensitivity to chemotherapy drugs and chemotherapy effects, and achieve the effect of high sensitivity

Inactive Publication Date: 2020-06-02
INST OF LAB ANIMAL SCI CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, new or improved chemotherapy drugs for colorectal cancer have been emerging, however, some patients are not sensitive to chemotherapy drugs, resulting in poor chemotherapy effect

Method used

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  • Application of Smurf1 gene, expression product and derivative or inhibitor thereof in chemotherapy of colorectal cancer
  • Application of Smurf1 gene, expression product and derivative or inhibitor thereof in chemotherapy of colorectal cancer
  • Application of Smurf1 gene, expression product and derivative or inhibitor thereof in chemotherapy of colorectal cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1: Knockdown of Smurf1 in HCT116 cells promotes apoptosis induced by Gemcitabine and Cisplatin

[0044] Stable HCT116 cells knocked down by Smurf1 were selected and plated in 12-well plates. After 24 hours, different concentrations of Gemcitabine (0.5, 1, 2, 5, 10 μM) or Cisplatin (12.5, 25, 50, 100, 200 μM) were applied to HCT116 cells for 48 hours. Afterwards, cell apoptosis was detected by flow cytometry and the percentage of apoptotic cells was statistically analyzed.

[0045] The results showed that after Smurf1 knockdown, the number of HCT116 apoptotic cells gradually increased with the increase of the working concentration of Gemcitabine ( figure 1 A). Similarly, after Smurf1 knockdown, the number of HCT116 apoptotic cells gradually increased with the increase of the working concentration of Cisplatin ( figure 1 B). The above results indicated that knockdown of Smurf1 in HCT116 cells promoted apoptosis induced by Gemcitabine and Cisplatin.

Embodiment 2

[0046] Example 2: Knocking down Smurf1 in HCT116 cells significantly enhanced the sensitivity of cells to Gemcitabine and Cisplatin ( Figure 4 )

[0047] The stable HCT116 cells with Smurf1 knockdown were selected and plated in 96-well plates. After 24 hours, different concentrations of Gemcitabine and Cisplatin (0, 0.5, 1, 2, 5, 10, 20, 50 μM) and the combination of the two (0 , 0.1, 0.2, 0.5, 1, 2, 5, 10μM) acted on HCT116 cells for 48h, and after CCK8 incubated the cells for 2h, the OD450nm was detected by a microplate reader, and the cell viability (cellviability, %) was statistically analyzed,

[0048] The results showed that Gemcitabine ( figure 2 A), Cisplatin ( figure 2 B) alone or in combination ( figure 2 C) Both can significantly reduce the survival rate of HCT116 cells knocked down by Smurf1, indicating that knocking down Smurf1 significantly enhances the sensitivity of HCT116 cells to Gemcitabine and Cisplatin.

Embodiment 3

[0049] Example 3: Tumor-bearing mice derived from HCT116 cells confirm that knocking down Smurf1 enhances the sensitivity of tumor cells to Gemcitabine and Cisplatin

[0050] In order to further confirm the effect of Smurf1 on the sensitivity of colorectal cancer cell HCT116 to chemotherapy drugs Gemcitabine and Cisplatin, in vivo experiments were carried out. For 6-8 week-old nude mice, each nude mouse was subcutaneously injected with 8×10 5 Control (HCT116 cells without Smurf1 knockdown) and HCT116 cells with Smurf1 knockdown, when the tumor grows to 100mm 3 Gemcitabine (50mg / kg), Cisplatin (2.5mg / kg) and the combination of the two (25mg / kgGemcitabine+1.25mg / kg Cisplatin) were administered intraperitoneally, once a week, and the mice were sacrificed after 4 weeks, and the tumor size to take pictures ( image 3 A) and weighing the tumor and performing statistical analysis ( image 3 B).

[0051] The results showed that Gemcitabine, Cisplatin and the combination of the two...

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Abstract

The invention belongs to the field of medicines and discloses a novel application of Smurf1, in particular to a novel application of a Smurf1 gene, an expression product and a derivative or an inhibitor thereof in chemotherapy of colorectal cancer. The influence of high and low level of the Smurf1 gene and the expression product on chemotherapeutic drug sensitivity of colorectal cancer cells is detected through cell, tumor-bearing mice and a PDX model, a result shows that low-expression colorectal cancers of Smurf1 have higher sensitivity to chemotherapeutic drugs. Therefore, the invention provides an application of the Smurf1 gene as well as the expression product and the derivative thereof in preparation of a marker showing sensitivity of the tumor cells to the chemotherapeutic drugs. The invention also provides an application of the Smurf1 inhibitor in preparation of a sensitizer of the tumor cells to the chemotherapeutic drugs.

Description

technical field [0001] The invention belongs to the field of medicine, and specifically relates to a new application of Smurf1, in particular to a new application of Smurf1 gene, expression product and its derivative or its inhibitor. Background technique [0002] The balance of protein synthesis and degradation in eukaryotic cells plays an important role in the maintenance of cellular homeostasis. There are two protein degradation pathways in eukaryotic cells, the lysosomal pathway and the ubiquitin-proteasome pathway, which operate in different ways. The lysosomal pathway mainly degrades exogenous proteins that enter the cell through endocytosis or pinocytosis, while the ubiquitin-proteasome pathway mainly degrades intracellular proteins. Enzyme, E1), ubiquitin-conjugating enzyme (ubiquitin-conjugating enzyme, E2) and ubiquitin ligase (ubiquitin ligase, E3) under the cascade action, the ubiquitin called "death tag" is marked, and thus is protease The body recognizes and ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886G01N33/574A61K45/00A61P35/00
CPCC12Q1/6886G01N33/57484G01N33/57419G01N33/57446A61K45/00A61P35/00C12Q2600/106C12Q2600/158
Inventor 韦荣飞郭静
Owner INST OF LAB ANIMAL SCI CHINESE ACAD OF MEDICAL SCI
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