"A novel in vivo model for the study of human breast cancer metastasis using primary breast tumor-initiating cells from patient biopsies"

Abstract

BackgroundThe study of breast cancer metastasis depends on the use of established breast cancer cell lines that do not accurately represent the heterogeneity and complexity of human breast tumors. A tumor model was developed using primary breast tumor-initiating cells isolated from patient core biopsies that would more accurately reflect human breast cancer metastasis.MethodsTumorspheres were isolated under serum-free culture conditions from core biopsies collected from five patients with clinical diagnosis of invasive ductal carcinoma (IDC). Isolated tumorspheres were transplanted into the mammary fat pad of NUDE mice to establish tumorigenicity in vivo. Tumors and metastatic lesions were analyzed by hematoxylin and eosin (H+E) staining and immunohistochemistry (IHC).ResultsTumorspheres were successfully isolated from all patient core biopsies, independent of the estrogen receptor α (ERα)/progesterone receptor (PR)/Her2/neu status or tumor grade. Each tumorsphere was estimated to contain 50-100 cells. Transplantation of 50 tumorspheres (1-5 × 103 cells) in combination with Matrigel into the mammary fat pad of NUDE mice resulted in small, palpable tumors that were sustained up to 12 months post-injection. Tumors were serially transplanted three times by re-isolation of tumorspheres from the tumors and injection into the mammary fat pad of NUDE mice. At 3 months post-injection, micrometastases to the lung, liver, kidneys, brain and femur were detected by measuring content of human chromosome 17. Visible macrometastases were detected in the lung, liver and kidneys by 6 months post-injection. Primary tumors variably expressed cytokeratins, Her2/neu, cytoplasmic E-cadherin, nuclear β catenin and fibronectin but were negative for ERα and vimentin. In lung and liver metastases, variable redistribution of E-cadherin and β catenin to the membrane of tumor cells was observed. ERα was re-expressed in lung metastatic cells in two of five samples.ConclusionsTumorspheres isolated under defined culture conditions from patient core biopsies were tumorigenic when transplanted into the mammary fat pad of NUDE mice, and metastasized to multiple mouse organs. Micrometastases in mouse organs demonstrated a dormancy period prior to outgrowth of macrometastases. The development of macrometastases with organ-specific phenotypic distinctions provides a superior model for the investigation of organ-specific effects on metastatic cancer cell survival and growth.

Treatment of Thyponium Flageliforme in Combination with Curcuma Zedoaria, and Phyllanthus Niruri Synergistically Enhances Apoptotic and Anti-Proliferative Effect on Breast Cancer

“…also pointed out that inoculation of breast cancer into femoral artery was able to stimulate multiple osteolytic lesions in the distal femoral and proximal tibiae within 18 days after inoculation, with a success rate of 95-100 % and no additional co morbidity 17 . Lastly, inoculation of tumor cells isolated from human suffer from breast cancer into mammary fat pad of nude mice was significantly induce breast cancer develop and metastasis to multiple mouse organs 18 Nevertheless, in the present study the tumor volume resulted from breast cancer inoculation in all groups were not similar (comparable) in size. The highest volume occurred in TCP-85, followed by C-G, and the lowest was in TCP-125 (Figure 2).…”

Rutin as A Novel c-Met Inhibitory Lead for The Control of Triple Negative Breast Malignancies

“…Hence, it was interesting to compare binding affinities and poses of rutin with oleocanthal at the c-Met kinase domain. The X-ray crystal structure PDB 4XYF was exploited to verify the stated aim (25). (–)-Oleocanthal showed excellent binding affinity at the c-Met's ATP kinase domain (Fig.…”

Extracellular vesicles from bone marrow mesenchymal stem/stromal cells transport tumor regulatory microRNA, proteins, and metabolites

“…Breast tumors from different groups were excised from mice for H&E staining as described previously [62]. …”