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GFP-labeled PNC breast cancer cell line with high metastatic and applications thereof

A technology of breast cancer cells and GFP-PTB, which is applied to cells modified by the introduction of foreign genetic material, recombinant DNA technology, measurement/testing of microorganisms, etc., can solve problems such as unclear mechanism of action, and achieve simple and easy screening steps The effect of quick operation and method

Inactive Publication Date: 2014-08-13
ZHEJIANG UNIV
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Problems solved by technology

However, as a complex component of traditional Chinese medicine, its specific mechanism of action is still unclear. At present, with the deepening of research on the anti-tumor mechanism of traditional Chinese medicine extracts, especially the anti-tumor mechanism of traditional Chinese medicine monomers, the prospect of traditional Chinese medicine monomers in tumor treatment has become increasingly popular. focus on

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  • GFP-labeled PNC breast cancer cell line with high metastatic and applications thereof

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Embodiment 1

[0040] One, MDA-MB-231 cells (purchased to ATCC), orthotopic inoculation in the mammary glands of female nude mice-separation of lymph node metastases-in vitro culture to obtain metastatic breast cancer cells-inoculation of mammary glands in situ again (repeated 5 times), obtained high The aggressive breast cancer cell line MDA-MB-231M;

[0041]2. Using PTB antibody (purchased from Invitrogen, diluted 1:1000) for immunofluorescence detection, the proportion of PNC in MDA-MB-231M cells was 95.7%;

[0042] 3. Construction of the PTB expression plasmid (GFP-PTB) with the GFP marker: the human PTB cDNA sequence was amplified by high-fidelity DNA polymerase PCR, and the amplified fragment was inserted into the GFP expression plasmid pEGFP-C1 at the HindIII and BamH1 sites ( Purchased from Shanghai Heyuan Biotechnology Co., Ltd.), the PTB fusion protein expression plasmid with GFP tag was obtained;

[0043] Four, the above-mentioned GFP-PTB fusion protein expression plasmid is pass...

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Abstract

The invention belongs to the biological technical field, relates to a GFP-labeled PNC breast cancer cell line with high metastatic and applications thereof, and also relates to an anti-metastatic traditional Chinese medicine monomer screening model based on tumor invasion novel target PNC and applications thereof. The invention aims to provide a technology platform for selective development of anti-metastatic traditional Chinese medicine monomer through establishing a medicine in-vitro screening model based on novel tumor metastasis molecular marker PNC. The GFP-labeled PNC breast cancer cell line with high metastatic is constructed by expressing a plasmid transfected high metastatic breast cancer cell line by a PFP-PTB fusion protein. The GFP-labeled PNC high metastatic breast cancer cell model does not need other in-vitro label, special reagents, or consumables, can be directly applied to anti-metastatic traditional Chinese medicine monomer screening, and has the advantages of simple screening steps, convenience, low cost, easy operation, and good stability.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a highly metastatic breast cancer cell line with GFP-labeled PNC and its application, and a screening model of an anti-metastatic traditional Chinese medicine monomer based on a new tumor aggressive target PNC and its application. Background technique [0002] The incidence of breast cancer in my country is increasing year by year, and the proportion of male breast cancer patients is also increasing. Although the clinical treatment of primary breast cancer by surgery, radiotherapy and chemotherapy has achieved initial success, the metastasis caused by advanced breast cancer is still an important life-threatening problem. At present, there is no clear and effective anti-metastatic drug. Traditional chemotherapeutic drugs such as adriamycin, daunorubicin, mitomycin, fluorouracil deoxynucleotide, etc., not only kill tumor cells, but also normal cells in the body. Cause serious injury, wi...

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Application Information

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IPC IPC(8): C12N5/10C12N15/85C12Q1/02
Inventor 刘艳宁陈智楼国华王静吴珊珊
Owner ZHEJIANG UNIV
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