New application of substance in regulation and control of YAP (Yes-associated protein) and/or TEAD and/or RHAMM (Receptor for Hyaluronan Mediated Motility) expression level

A technology of expression level and substance, applied in DNA/RNA fragments, cells modified by introducing foreign genetic material, application, etc., can solve the problems of limited efficacy of radiotherapy and chemotherapy

Active Publication Date: 2014-04-02
SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional surgery, radiotherapy and chemotherapy currently used in the treatment of breast cancer, especially advanced breast cancer, have limited efficacy, and breast cancer still ranks first in the list of female cancer killers

Method used

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  • New application of substance in regulation and control of YAP (Yes-associated protein) and/or TEAD and/or RHAMM (Receptor for Hyaluronan Mediated Motility) expression level
  • New application of substance in regulation and control of YAP (Yes-associated protein) and/or TEAD and/or RHAMM (Receptor for Hyaluronan Mediated Motility) expression level
  • New application of substance in regulation and control of YAP (Yes-associated protein) and/or TEAD and/or RHAMM (Receptor for Hyaluronan Mediated Motility) expression level

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0093] Example 1, RHAMM plays an important role in the migration and invasion of breast cancer cells, and the knockdown of RHAMM mediated by RNAi can inhibit the migration and invasion of breast cancer cells

[0094] 1. ShRNA targeting RHAMM can significantly reduce the expression of RHAMM protein in MDA-MB-231 cells

[0095] First package a lentivirus (named shRHAMM1 lentivirus) expressing shRNA targeting RHAMM (named shRHAMM1), package another lentivirus (named shRHAMM2 lentivirus) expressing shRNA targeting RHAMM (named shRHAMM2), A lentivirus (named NT lentivirus) expressing random sequence shRNA (named shC) was packaged as a negative control. Then the triple-negative breast cancer cell line MDA-MB-231 was infected with shRHAMM1 lentivirus, shRHAMM2 lentivirus and NT lentivirus separately. After 12 hours, the virus was removed, and the virus-infected MDA-MB-231 was cultured normally. MB-231 cells. The results showed that the RHAMM content in the MDA-MB-231 cell line infe...

Embodiment 2

[0109] Example 2, YAP and TEAD play an important role in the expression of RHAMM, and the knockdown of RNAi-mediated YAP or TEAD inhibits the migration and invasion of breast cancer cells by reducing the protein level of RHAMM

[0110] 1. ShRNA targeting YAP and shRNA targeting TEAD can significantly reduce the expression of RHAMM protein in MDA-MB-231 cells

[0111] First, package a lentivirus (named shYAP lentivirus) expressing shRNA targeting YAP (named shYAP), package a lentivirus (named shTEAD lentivirus) expressing shRNA (named shTEAD) targeting TEAD1 / 3 / 4 virus), and a lentivirus (named NT lentivirus) that packaged a shRNA expressing a random sequence (named shC) was used as a negative control. Then the triple-negative breast cancer cell line MDA-MB-231 was infected with shYAP lentivirus, shTEAD lentivirus and NT lentivirus separately. After 12 hours, the virus was removed, and the virus-infected MDA-MB-231 was cultured normally. MB-231 cells. The results showed that t...

Embodiment 3

[0126] Example 3. High expression of YAP promotes transcription and translation of RHAMM.

[0127] Use Lipofectamine2000 transfection reagent (Invitrogen) to introduce the YAP expression plasmid V5-YAP plasmid (the YAP coding gene is inserted between the EcoRI and NotI sites of pcDNA3.1V5 / His (Invitrogen) to obtain the V5-YAP expression plasmid) into 293T cells to obtain YAP The recombinant cells with protein overexpression were named as 293T-YAP cells. Extract the total RNA of 293T cells and 293T-YAP cells, and detect the transcription level of RHAMM by fluorescence quantitative method according to the method in 1.3, that is, the change of the mRNA level of RHAMM; - Changes in protein levels of RHAMM in YAP cells. V5 antibody was purchased from GeneTeX Company.

[0128] The results showed that the mRNA level of RHAMM in 293T-YAP cells was 3.3 times that of RHAMM in 293T cells, and the protein level of RHAMM in 293T-YAP cells was 1.55 times that of RHAMM in 293T cells ( im...

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Abstract

The invention discloses a new application of substance in regulation and control of YAP and / or TEAD and / or RHAMM expression level. The new application provided by the invention is application of substance for regulating and controlling YAP expression level in preparation of product for regulating and controlling RHAMM expression level in animal cells. Experiment of the invention shows that YAP and TEAD both are new targets for interfering RHAMM expression, RHAMM expression can be reduced by reducing expression of YAP or TEAD, reduction of YAP, TEAD or RHAMM expression can restrain transfer ability and invasive ability of triple negative breast cancer cell line MDA-MB-231, different approaches can be used for interfering, regulating and controlling any point from Meva to YAP in RHAMM, or YAP / TEAD, or Mevalonate application, and thus, interference on cell migration and treatment of tumor metastasis can be realized.

Description

technical field [0001] The present invention relates to a new application of substances for regulating the expression level of YAP and / or TEAD and / or RHAMM. Background technique [0002] Breast cancer is the most common tumor among women in both developed and developing countries. Worldwide, 1.2 million women suffer from breast cancer every year, and 500,000 women die from breast cancer. In developed countries in Europe and the United States, 1 in every 8-9 people suffers from breast cancer. In developing countries, the incidence of breast cancer has increased dramatically due to increasing life expectancy, urbanization and adoption of Western lifestyles. In my country, the statistics of Beijing, Shanghai, Tianjin, Guangzhou, Shenzhen and other big cities show that breast cancer is also the most common malignant tumor among women in my country, and the incidence rate is increasing year by year. Data show that the incidence of female breast cancer in my country is increasi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/867C12N5/10
Inventor 黄来强王中原吴彦萍王海峰张扬清
Owner SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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