Assay of Gcdfp-15 by Elisa: An Available Method for in Vitro Studies of Functional Differentiation in Human Breast Cancer

An enzyme-linked immunosorbent assay (ELISA) was applied to a light protein, isolated from human breast cyst fluid (BCF) termed « gross cystic disease fluid protein - 15 Kda » (GCDFP-15), a potential differentiation marker in in vitro human breast cancer studies. The detection limits of this procedure, performed in microtiter plates, were 0.5 to 250 ng/well corresponding to 10 ng/ml to 5 ng/ml of sample or antigen solution. Possible cross-reaction with various antigens, especially those found in culture media, were investigated. The correlation coefficient between enzymoassay and radioimmunoassay was 0.978. The results showed that quantification of GCDFP-15 by ELISA is a specific and highly sensitive method. This procedure may be of interest in in vitro studies on the functional differentiation of breast cancer cells.

Download Full-text

Can Animal and In Vitro Studies Give New, Relevant Answers to Questions Concerning Mammographic Screening for Human Breast Cancer?23

JNCI Journal of the National Cancer Institute ◽

10.1093/jnci/61.6.1537 ◽

1978 ◽

Keyword(s):

Breast Cancer ◽

Human Breast Cancer ◽

Mammographic Screening ◽

In Vitro Studies ◽

Human Breast

Download Full-text

In vitro studies of phospholipid-modified PAMAM-siMDR1 complexes for the reversal of multidrug resistance in human breast cancer cells

International Journal of Pharmaceutics ◽

10.1016/j.ijpharm.2017.06.026 ◽

2017 ◽

Vol 530 (1-2) ◽

pp. 291-299 ◽

Cited By ~ 16

Author(s):

Jing Liu ◽

Jun Li ◽

Nan Liu ◽

Nana Guo ◽

Chen Gao ◽

...

Keyword(s):

Breast Cancer ◽

Multidrug Resistance ◽

Cancer Cells ◽

Human Breast Cancer ◽

Breast Cancer Cells ◽

In Vitro Studies ◽

Human Breast Cancer Cells ◽

Human Breast

Download Full-text

Wortmannin Reduces Metastasis and Angiogenesis of Human Breast Cancer Cells via Nuclear Factor-κB-Dependent Matrix Metalloproteinase-9 and Interleukin-8 Pathways

Journal of International Medical Research ◽

10.1177/147323001204000305 ◽

2012 ◽

Vol 40 (3) ◽

pp. 867-876 ◽

Cited By ~ 9

Author(s):

J Li ◽

F Li ◽

H Wang ◽

X Wang ◽

Y Jiang ◽

...

Keyword(s):

Breast Cancer ◽

Matrix Metalloproteinase ◽

Cancer Cells ◽

Human Breast Cancer ◽

Breast Cancer Cells ◽

Enzyme Linked Immunosorbent Assay ◽

Nuclear Factor ◽

Human Breast

OBJECTIVE: To investigate whether inhibition of Akt phosphorylation by the phosphatidylinositol 3-kinase (PI3K) inhibitor, wortmannin, reduces metastasis and angiogenesis in a human breast cancer cell line via nuclear factor (NF)-κB-dependent matrix metalloproteinase (MMP)-9 and interleukin (IL)-8 pathways. METHODS: MDA-MB-231 cells were treated with wortmannin 0 – 200 nM for 4 h. Restoration of Akt activity was evaluated by transfection of cells with constitutively active myristoylated Akt (myr-Akt). NF-κB, MMP-9 and IL-8 proteins were detected by electrophoretic mobility shift assay, Western blot or enzyme-linked immunosorbent assay. The chicken embryo chorio-allantoic membrane assay, cell motility and migration assays were used to evaluate angiogenesis and invasion in vitro. A mouse pseudo metastatic breast cancer model was used to assess the effects of wortmannin on metastasis and angiogenesis in vivo. RESULTS: Wortmannin inhibited the phosphorylation of Akt, upregulation of NF-κB, MMP-9, IL-8, and in vitro cell invasion and angiogenesis, in a dose-dependent manner. Transfection of myr-Akt reversed the cellular and biochemical effects of wortmannin in vitro. Wortmannin also significantly inhibited tumour metastasis and angiogenesis in vivo. CONCLUSION: The findings of the present study suggest that wortmannin inhibits metastasis and angiogenesis in breast cancer cells via PI3K/Akt/NF-κB-mediated MMP-9 and IL-8 signalling pathways.

Download Full-text