Fusion of apoptosis‐related protein Cytochrome c with anti‐HER‐2 single‐chain antibody targets the suppression of HER‐2+ breast cancer

Triple-negative breast cancer (TNBC), the most aggressive subtype, does not respond to targeted therapy due to the lack of hormone receptors. There is an urgent need for alternative therapies, including natural product-based anti-cancer drugs, at lower cost. We investigated the impact of a Calligonum comosum L’Hér. methanolic extract (CcME) on the TNBC MDA-MB-231 cell line proliferation and related cell death mechanisms performing cell viability and cytotoxicity assays, flow cytometry to detect apoptosis and cell cycle analysis. The apoptosis-related protein array and cellular reactive oxygen species (ROS) assay were also carried out. We showed that the CcME inhibited the TNBC cell viability, in a dose-dependent manner, with low cytotoxic effects. The CcME-treated TNBC cells underwent apoptosis, associated with a concomitant increase of apoptosis-related protein expression, including cytochrome c, cleaved caspase-3, cyclin-dependent kinase inhibitor p21, and the anti-oxidant enzyme catalase, compared with the untreated cells. The CcME also enhanced the mitochondrial transition pore opening activity and induced G0/G1 cell growth arrest, which confirmed the cytochrome c release and the increase of the p21 expression detected in the CcME-treated TNBC cells. The CcME-treated TNBC cells resulted in intracellular ROS production, which, when blocked with a ROS scavenger, did not reduce the CcME-induced apoptosis. In conclusion, CcME exerts anti-proliferative effects against TNBC cells through the induction of apoptosis and cell growth arrest. In vivo studies are justified to verify the CcME anti-proliferative activities and to investigate any potential anti-metastatic activities of CcME against TNBC development and progression.

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Anti-Her2 single-chain antibody mediated DNMTs-siRNA delivery for targeted breast cancer therapy

Journal of Controlled Release ◽

10.1016/j.jconrel.2012.05.015 ◽

2012 ◽

Vol 161 (3) ◽

pp. 875-883 ◽

Cited By ~ 28

Author(s):

Shuang Dou ◽

Yan-Dan Yao ◽

Xian-Zhu Yang ◽

Tian-Meng Sun ◽

Cheng-Qiong Mao ◽

...

Keyword(s):

Breast Cancer ◽

Cancer Therapy ◽

Sirna Delivery ◽

Single Chain ◽

Breast Cancer Therapy ◽

Single Chain Antibody

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A vaccine approach for breast cancer immunotherapy by targeting HER2/neu antigen to B cells via CD19 molecule.

Journal of Clinical Oncology ◽

10.1200/jco.2011.29.27_suppl.287 ◽

2011 ◽

Vol 29 (27_suppl) ◽

pp. 287-287

Author(s):

D. Xiang ◽

Y. Ma ◽

C. Ding ◽

G. H. Kloecker ◽

J. Yan

Keyword(s):

Breast Cancer ◽

B Cells ◽

Fusion Protein ◽

Tumor Cells ◽

Fusion Proteins ◽

In Vivo Studies ◽

Single Chain ◽

Tumor Vaccines ◽

Single Chain Fv ◽

Her 2

287 Background: Trastuzumab has been proven to be effective in the immunotherapy for Her-2/neu-positive breast cancer. However, this adoptively transferred therapeutic Ab must be continuously given to the patient with huge financial cost. Thus, it would be desirable if tumor vaccines could elicit long-lasting trastuzumab-like Ab. Methods: Constructing fusion protein of anti-mouse CD19 single chain variable fragment (anti-CD19-scFv) with human Her-2/neu extracellular domain P3-4 (P3-4) by extracting total RNA from Rat anti-mouse CD19 hybridoma (1D3) and first strand cDNA was synthesized accordingly. VH and VL were amplified using designed primers. Single chain Fv (VL-VH) was then synthesized by overlapping PCR. Then constructs were sequenced and cloned into pET-20b(+) vectors. Fragments P3-4 of extracellular domains of human Her-2/neu was cloned from pcDNA3.1-Her2 and then cloned into pET-20b(+)-scFv. Series of constructs were expressed and purified according to standard protocol and verified by Western Blot. Results: Both in vitro and in vivo studies demonstrated that fusion proteins anti-CD19-scFv and anti-CD19-scFv-P3-4 but not P3-4 could specifically bind to B cells with high affinity. Mice immunized with anti-CD19-scFv-P3-4 secreted higher titers of IgG and IgM than those from controls (p<0.05). Studies also demonstrated that sera from anti-CD19-scFv-P3-4 but not anti-CD19-scFv or P3-4 immunized mice stained with Her-2/neu expressing SKOV-3 tumor cells. These Abs also competitively inhibited trastuzumab-mediated Ag binding, suggesting that trastuzumab-like Ab responses were elicited. WT mice immunized with anti-CD19-scFv-P3-4 fusion protein then challenged with D2/F2-Her-2 mammary tumor cells showed significantly reduced tumor burden compared to those immunized with control fusion proteins (p<0.05) and had enhanced median overall survival to 45 days versus 34 days in WT mice immunized with either anti-CD19-scFv or P3-4. Conclusions: Targeting of Her-2/neu antigens to B cells stimulates Th-dependent humoral immune responses with anti tumor effect in mouse model. These findings provide a novel avenue for successful development of breast cancer vaccination strategy and help to fight for cancer.

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Antiproliferative and apoptotic effects of a specific anti-insulin-like growth factor I receptor single chain antibody on breast cancer cells

Tumor Biology ◽

10.1007/s13277-016-5323-4 ◽

2016 ◽

Vol 37 (11) ◽

pp. 14841-14850 ◽

Cited By ~ 4

Author(s):

Morteza Motallebnezhad ◽

Vahid Younesi ◽

Leili Aghebati-Maleki ◽

Hamid Nickho ◽

Elham Safarzadeh ◽

...

Keyword(s):

Breast Cancer ◽

Growth Factor ◽

Cancer Cells ◽

Breast Cancer Cells ◽

Insulin Like Growth Factor ◽

Single Chain ◽

Single Chain Antibody ◽

Factor I ◽

Growth Factor I ◽

Apoptotic Effects

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The FGF ‐1‐specific single‐chain antibody scFv1C9 effectively inhibits breast cancer tumour growth and metastasis

Journal of Cellular and Molecular Medicine ◽

10.1111/jcmm.12371 ◽

2014 ◽

Vol 18 (10) ◽

pp. 2061-2070 ◽

Cited By ~ 6

Author(s):

Hengliang Shi ◽

Chunling Fu ◽

Wei Wang ◽

Yu Li ◽

Shuang Du ◽

...

Keyword(s):

Breast Cancer ◽

Tumour Growth ◽

Single Chain ◽

Single Chain Antibody

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Extracellular cytochrome c, a mitochondrial apoptosis-related protein, induces arthritis

Rheumatology ◽

10.1093/rheumatology/keh406 ◽

2004 ◽

Vol 44 (1) ◽

pp. 32-39 ◽

Cited By ~ 52

Author(s):

R. Pullerits ◽

M. Bokarewa ◽

I.-M. Jonsson ◽

M. Verdrengh ◽

A. Tarkowski

Keyword(s):

Cytochrome C ◽

Mitochondrial Apoptosis ◽

Related Protein ◽

Apoptosis Related Protein

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Anti-Proliferative Effect of Specific Anti-EGFR Single Chain Antibody on Triple Negative Breast Cancer Cells

Reports of Biochemistry and Molecular Biology ◽

10.29252/rbmb.9.2.180 ◽

2020 ◽

Vol 9 (2) ◽

pp. 180-187

Author(s):

Foroogh Abdollahzadeh ◽

Forough Nejatollahi ◽

◽

Keyword(s):

Breast Cancer ◽

Cancer Cells ◽

Triple Negative Breast Cancer ◽

Breast Cancer Cells ◽

Triple Negative ◽

Single Chain ◽

Single Chain Antibody ◽

Proliferative Effect

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Nobiletin enhances Pirarubicin chemosensitivity of breast cancer cell MDA-MB-231 by regulating PER2

E3S Web of Conferences ◽

10.1051/e3sconf/202123302010 ◽

2021 ◽

Vol 233 ◽

pp. 02010

Author(s):

Fahu Yuan ◽

Xinghua Liao ◽

Tongcun Zhang

Keyword(s):

Breast Cancer ◽

Cell Cycle ◽

Scratch Test ◽

Inhibitory Effect ◽

Control Group ◽

Potential Mechanism ◽

Related Protein ◽

Repair Ability ◽

Apoptosis Related Protein ◽

Related Proteins

To explore the potential mechanism of nobiletin in improving the sensitivity of breast cancer MDA-MB-231 cells to Pirarubicin. MDA-MB-231 cells were randomly divided into Control group, Nobiletin group, Pirarubicin group and Nobiletin co-Pirarubicin group. CCK-8 assay was used to detect the inhibitory effect of Nobiletin co-Pirarubicin on the proliferation of MDA-MB-231 cells, and the cell cycle and apoptosis of MDA-MB-231 cells were detected by flow cytometry. The effect of nobiletin combined with Pirarubicin on the migration and repair ability of MDA-MB-231 cells was detected by scratch test. Western blotting was used to determine the effect of nobiletin co-Pirarubicin on Period2 (PER2), apoptosis-related proteins Bax and BCL-2 expression levels in MDA-MB-231 cells. The results showed that the sensitivity of MDA-MB-231 cells to Pirarubicin was enhanced by nobiletin. Compared with Pirarubicin or nobiletin alone, Pirarubicin combined with nobiletin promoted the apoptosis of MDA-MB-231 cells and the expression of apoptosis-related protein Bax, and reduced the expression of apoptosis-inhibiting protein BCL-2. The combination of nobiletin and Pirarubicin significantly inhibited the proliferation and promoted the apoptosis of MDA-MB-231 cells, which may be mediated by the enhancement of PER2 expression by nobiletin.

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