Application of metadherin (MTDH) gene vaccine in preparation of breast cancer cell growth inhibitor

A gene vaccine and breast tumor technology, applied in the field of preparation of breast cancer growth inhibitors, can solve the problems of limited breast cancer methods and effects, and achieve the effects of inhibiting lung metastasis and prolonging lifespan

Inactive Publication Date: 2012-02-01
上海宜诺迪生物科技有限公司
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

Based on the current means and effects of breast cancer treatment are limited, researchers in this field believe that t

Method used

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  • Application of metadherin (MTDH) gene vaccine in preparation of breast cancer cell growth inhibitor
  • Application of metadherin (MTDH) gene vaccine in preparation of breast cancer cell growth inhibitor
  • Application of metadherin (MTDH) gene vaccine in preparation of breast cancer cell growth inhibitor

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Experimental program
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Effect test

Embodiment 1

[0066] Materials and Methods:

[0067] 1. Sources of animals, bacterial strains and cell lines

[0068] 6-8 week old female BALB / c mice were purchased from Shanghai Experimental Animal Center. The mice were kept in accordance with the guidelines for the use of experimental animals of the Shanghai Jiao Tong University School of Medicine. Attenuated Salmonella strain (Salmonella typhimurium, dam - and AroA - ) RE88 and mouse 4T1 breast cancer cell lines were kindly provided by Prof. Ralph A. Reisfeld (The Scripps Research Institute, La Jolla, CA).

[0069] 2. Cell culture

[0070] The mouse breast cancer cell line 4T1 was cultured with RMPI-1640 medium (HyClone, Logan, UT) supplemented with 10% fetal bovine serum (HyClone), 100 units / mL penicillin and 100 ug / mL streptomycin (Invitrogen, Carlsbad, CA) . Human embryonic kidney cell line HEK293T with DMEM (HyClone) supplemented with 10% fetal bovine serum, penicillin and streptomycin at 37°C, 5% CO 2 cultured in an incubator...

Embodiment 2

[0156] Example 2, MTDH expression profile analysis in mouse tissues and 4T1 breast cancer cell lines

[0157] It has been confirmed in the literature that MTDH is highly expressed in most breast cancer patients [10], so it may be used as a target for the treatment of breast cancer. Since the tumor vaccine target must have two basic conditions: high expression in tumors and low expression in normal tissues, in order to verify whether the MTDH gene vaccine is feasible for the treatment of breast cancer, firstly, the expression of MTDH in mouse tissues and the mouse breast cancer cell line 4T1 was tested. The expression profile was analyzed. The results showed that the expression level of MTDH protein in 4T1 cell line and liver was higher, and the expression level in heart, spleen, kidney, lung, brain and small intestine was very low ( figure 2 ). Quantitative PCR results also showed that the expression level of MTDH gene mRNA in mouse liver and 4T1 cells was significantly hig...

Embodiment 3

[0158] Embodiment 3, the construction of MTDH gene vaccine vector and the detection of its protein expression

[0159] The pcDNA3.1 / Ub-Myc vector was selected as the gene vaccine expression vector, and the full-length cDNA of the mouse MTDH gene was subcloned into the expression vector ( Figure 4 ). Previous research results have shown that after the N-terminal of the target protein is coupled with ubiquitin protein, it can effectively improve the type I MHC antigen presentation pathway of the target protein[4,7]. Based on the mechanism of ubiquitin-mediated protein degradation, ubiquitin-conjugated target proteins can increase the efficiency of protein degradation through the proteasome, and such ubiquitin-conjugated vaccines can induce stronger CD8+ T cell-mediated activation than vaccines lacking ubiquitin. induced immune response. The MTDH vaccine plasmid or empty vector was transfected into HEK293T cells, and its expression was detected by immunoblotting ( Figure 5 )...

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Abstract

The invention belongs to the field of biotechnologies, and relates to a gene vaccine capable of inhibiting cancer cell growth. The gene vaccine can express a cell membrane surface protein metadherin (MTDH) and thus being named as a MTDH gene vaccine. The MTDH gene vaccine is utilized for immunization in an oral way. The MTDH gene vaccine can activate an immune response of CD8+ cytotoxic T cells on breast cancer cells 4T1 expressing MTDH antigens. The MTDH gene vaccine can inhibit breast cancer cell growth and pulmonary metastasis. Through being combined with adriamycin, the MTDH gene vaccine can improve adriamycin effects of inhibiting breast cancer cell growth and pulmonary metastasis, and prolong a life of a mouse loaded with cancer. More significantly, the MTDH gene vaccine does not produce obvious damages on main organs of a human body. Therefore, the MTDH gene vaccine provides a novel approach for improvement of the existing breast cancer treatment method.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to an MTDH gene vaccine and the application of the MTDH gene vaccine in preparing breast cancer growth inhibitors. Background technique [0002] Breast cancer is one of the most common types of cancer in women, and its death rate is reported to be the first among all tumors. In 2009, approximately 192,370 people were diagnosed with breast cancer in the United States, and 40,170 of them died of breast cancer [1]. It has also been reported that more than 40% of female patients with breast cancer are diagnosed with tumor metastasis [2]. The current treatment options for breast cancer include surgical resection of the primary lesion, regional lymph node dissection, hormone therapy, and chemotherapy and radiotherapy. However, even after successful surgical resection of the primary lesion and adjuvant chemotherapy, the recurrence rate and mortality rate of breast cancer are still high. Accord...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K39/00A61P35/00A61K31/704
Inventor 郭方朱晗
Owner 上海宜诺迪生物科技有限公司
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