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Application of NOD1 to preparation of products for inhibiting tumor SRC signal path

A signal pathway and product technology, applied in the direction of antineoplastic drugs, medical preparations containing active ingredients, pharmaceutical formulations, etc.

Inactive Publication Date: 2019-03-12
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports on the application of NOD1 in tumor therapy.

Method used

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  • Application of NOD1 to preparation of products for inhibiting tumor SRC signal path
  • Application of NOD1 to preparation of products for inhibiting tumor SRC signal path
  • Application of NOD1 to preparation of products for inhibiting tumor SRC signal path

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1 Effect of NOD1 expression on liver cancer

[0057] (1) SMMC7721 and BEL7402 liver cancer cells were treated with 3x10 5 Inoculate in a 6-well plate at a density of / ml, add 2ml of cell suspension to each well, and culture overnight. When the cell density was 60%-80%, the liver cancer cells were transfected with Si-NOD1, and the Si-NC transfection group was used as a control, and a blank control group was also set up. Si-NOD1 sequence: Sense: 5'-CCAGCUCGUUCAGAGCAAATT-3', as shown in SEQ ID NO.2; Anti-sense: 5'-UUUGCUCUGAACGAGCUGGTT-3', as shown in SEQ ID NO.3. Si-NC sequence: Sense: 5'-UUCUCCGAACGUGUCACGUTT, as shown in SEQ ID NO.4; Anti-sense: ACGUGACACGUUCGGAGAATT, as shown in SEQ ID NO.5.

[0058] Successfully interfered with the expression of NOD1 in the liver cancer cell lines BEL7402 and SMMC7721 ( figure 1 A), CCK-8 method detects that the proliferation activity of liver cancer cells is significantly improved ( figure 1 B); After inhibiting the expre...

Embodiment 2

[0062] Example 2 NOD1 inhibits MAPK signaling pathway

[0063] After interfering with NOD1 expression in liver cancer cell lines BEL7402 and SMMC7721, it was detected that the expression of cell cycle negative regulator protein P21 was significantly inhibited, while the expression levels of cell cycle activation proteins p-CyclinD, p-CyclinE and p-CDC2 were significantly up-regulated; Conversely, in the liver cancer cell lines HUH7 and HepG2, exogenous overexpression of NOD1 can significantly increase the expression of P21, while the expression levels of cell cycle activation proteins p-CylinD and p-CyclinE are significantly down-regulated; the above results further clarified that NOD1 can Negatively regulate the cell cycle of liver cancer cells. The further study of the present invention found that interfering with the expression of NOD1 in liver cancer cell lines can significantly activate the MAPK signaling pathway, on the contrary, overexpressing NOD1 can significantly inh...

Embodiment 3

[0064] Example 3 Screening of molecular targets for NOD1 to exert tumor suppressor effect in liver cancer cells

[0065] Co-immunoprecipitation method detected the endogenous and exogenous combination of NOD1 and SRC ( Figure 4 A-B). Immunofluorescence staining proved that NOD1 and SRC co-localized in the cytoplasm ( Figure 4 C) In vitro protein translation and co-immunoprecipitation methods prove that there is a direct binding effect between NOD1 and SRC ( Figure 4 D). In the further study, the present invention constructed the truncated mutants of NOD1 and SRC respectively, and proved that NOD1 combined with the SH3 domain of SRC through its CARD domain through co-immunoprecipitation method.

[0066] In the present invention, by interfering with the expression of NOD1 in liver cancer cells BEL7402, it is detected that inhibiting the expression of NOD1 can significantly increase the expression level of p-SRC without affecting the expression of SRC ( Figure 5 A); Conve...

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Abstract

The invention belongs to the field of biological medicines, and particularly relates to application of NOD1 to preparation of products for inhibiting MAPK signal path. The products have the at least one of the following functions from A1) to A7): A1) combination with SRC; A2) inhibition of expression of p-SRC; A3) inhibition of MAPK signal path; A4) inhibition of expression of p-ERK; A5) inhibition of expression of p-CyclinD; A6) inhibition of expression of p-CyclinE; and A7) promotion of expression of p21. The invention firstly discovers NOD1 can inhibit p-SRC through target binding of SRC and further inhibit the MAPK signal path. The discovery provides a new target spot for screening drugs for treating diseases related to abnormal MAPK signal path.

Description

technical field [0001] The invention belongs to the field of biomedicine, and specifically relates to the application of NOD1 in the preparation of products for inhibiting tumor SRC signaling pathways. Background technique [0002] Hepatocellular carcinoma (HCC) is the fifth most common cancer in the world, and its death rate ranks third. About 700,000 people die from HCC every year worldwide. As a large HCC country, our country accounts for about 40% of the world's liver cancer patients, and HCC is the second fatal tumor in our country. [0003] At present, the treatment for HCC is mainly based on surgical resection of tumor lesions. Although surgery prolongs the life of some patients, HCC still presents a high postoperative recurrence rate and metastasis rate, and is accompanied by a poor prognosis. The high recurrence rate of liver cancer metastasis is the main reason for the high mortality rate of HCC. At present, the main clinical measures for the recurrence and metas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K31/7088A61P35/00A61P1/16
CPCA61K31/7088A61K45/00A61P1/16A61P35/00
Inventor 韩丽辉马小敏邱驭旻朱礼慧李涛赵云雪林月轲马大鹏秦振志孙偲瑜沈雪城
Owner SHANDONG UNIV
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