Application of PITPNM3 gene in preparing medicine for inhibiting breast cancer invasion and transfer

A breast cancer, drug technology, applied in application, gene therapy, drug combination and other directions, can solve problems such as hindering the development of breast cancer migration methods

Inactive Publication Date: 2011-08-24
SUN YAT SEN MEMORIAL HOSPITAL SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the target receptor of CCL18 has not been identified, the further development of targeting CCL18 and its downstream signaling pathways to inhibit breast cancer migration has been hindered.

Method used

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  • Application of PITPNM3 gene in preparing medicine for inhibiting breast cancer invasion and transfer
  • Application of PITPNM3 gene in preparing medicine for inhibiting breast cancer invasion and transfer
  • Application of PITPNM3 gene in preparing medicine for inhibiting breast cancer invasion and transfer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1: The experiment of CCL18 promoting the invasion and migration of breast cancer

[0053] To investigate the function of CCL18, we designed a scratch assay to evaluate the effect of secreting and expressing CCL18 macrophages on the invasion and migration of breast cancer. Specifically, the experimental method is as follows: culture breast cancer cells at a density of 90%, use a pipette tip to draw a straight line on the cell monolayer, put the nest of selectively activated macrophages that secrete CCL18 into the culture system, add DMEMF12 Under the culture conditions of 0.4% BSA, B2720ul / ml, and EGF insulin, they were co-cultured for 12 hrs. The pictures of 0h and 12h were taken under a phase-contrast microscope, and the distances on both sides of the scratches at 0h and 12h on the pictures were analyzed by Carnoy software (Biovolution), and the percentage of the cell migration distance and the non-migration distance was calculated. It was proved that breast c...

Embodiment 2

[0054] Example 2: Experiment to prove that PITPNM3 is a transmembrane receptor activated by CCL18

[0055] In order to identify whether there is CCL18 receptor on the breast cancer cell membrane, MDA-MB-231 cells were incubated with CCL18 at 4°C for 3 h, fixed, and stained with Alexa488-labeled CCL18 antibody. Immunofluorescence staining demonstrated that CCL18 was present on the membrane of breast cancer cells ( image 3 A), suggesting that there are special protein molecules on the surface of breast cancer cells that can bind to CCL18 and mediate the biological effects of CCL18. In addition, adding a neutralizing antibody to CCL18 during the incubation of CCL18 with cells can prevent CCL18 from binding to breast cancer cell surface proteins ( image 3 B).

[0056] In order to identify unknown functional receptors of CCL18, we first treated breast cancer MDA-MB-231 cells with CCL18 protein, then extracted cell membrane proteins, and co-immunoprecipitated the membrane protei...

Embodiment 3

[0060] Example 3 PITPNM3 mediates CCL18 to promote breast cancer invasion and migration

[0061] In order to further explore the CCL18-PITPNM3 interaction in breast cancer cells, we demonstrated that silencing the expression of PITPNM3 with siRNA would affect the effect of CCL18 on breast cancer cells. The siRNA sequences are: PITPNM3 siRNA-1: sense strand 5′-GGGAGAAGUGGCUUCGUAATT-3′ antisense strand: 5′-UUACGAAGCCACUUCUCCCGG-3′; PITPNM3 siRNA-2: sense strand 5′-CGCGCAUGAUCCUGCGCAATT-3′ antisense strand: 5′- UUGCGCAGGAUCAUGCGCGAG-3'. Silencing the expression of PITPNM3 in breast cancer cells can significantly inhibit the role of CCL18 in promoting the migration of breast cancer cells to the level of breast cancer cell migration without the effect of CCL18 (P Figure 8 ). Transfection of breast cancer cells without rCCL18 effect with PITPNM3-siRNA did not affect the cell invasion and migration function (P>0.05, Figure 8 ). These results indicate that the biological function ...

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Abstract

The invention provides application of PITPNM3 gene in preparing medicines for inhibiting breast cancer invasion and transfer. CCL18 promotes breast cancer invasion and transfer, and the PITPNM3 is identified as a CCL18 receptor and is a new target for inhibiting breast cancer invasion and transfer. The medicine for inhibiting breast cancer invasion and transfer takes a medicine for inhibiting the PITPNM3 gene in a targeted way as an active ingredient. The medicine for inhibiting the PITPNM3 gene in the targeted way takes siRNA expressed by PITPNM3 or / and a G protein coupling receptor inhibitor of pertussis toxin as active ingredients to inhibit the biological effect caused by combination of the CCL18 and the PITPNM3. The invention provides the CCL18 to promote the breast cancer invasion and transfer through the PITPNM3 for the first time, the PITPNM3 is the receptor of the CCL18, and the PITPNM3 receptor can be inhibited in the targeted way so as to inhibit the breast cancer invasion and transfer.

Description

technical field [0001] The invention relates to the application of PITPNM3 gene in the preparation of drugs for inhibiting the invasion and metastasis of breast cancer. Background technique [0002] Breast cancer metastasis is the root cause of treatment failure for patients, but the exact mechanism of tumor metastasis has not been systematically elucidated, and there is no effective method to inhibit breast cancer metastasis. Research on the mechanism of promoting breast cancer metastasis and establish molecular targets to inhibit breast cancer metastasis , is an effective way to improve the prognosis of breast cancer. There are a variety of metastasis-related stromal cells in the tumor microenvironment, and a large number of clinical studies have proved that tumor-associated macrophages (TAM) can promote the invasion and migration of various tumors, including breast cancer. A large number of studies have proved that TAM promotes the invasion and metastasis of breast cance...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K38/16C12N15/12A61P35/00A61P35/02
Inventor 宋尔卫陈静琦姚雪彪
Owner SUN YAT SEN MEMORIAL HOSPITAL SUN YAT SEN UNIV
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