Fangjihuangqi Decoction inhibits MDA‐MB‐231 cell invasion in vitro and decreases tumor growth and metastasis in triple‐negative breast cancer xenografts tumor zebrafish model

Abstract

Triple‐negative breast cancer (TNBC) is a basal‐like cancer which is considered to be more intrusive, have a poorer prognosis and chemoresistance. TNBC is characterized by the presence of epithelial to mesenchymal transition (EMT) that plays a major role in the progression of the cancer. In the present study, we first use a classic prescription of Chinese medicine Fangjihuangqi Decoction to treat TGFβ1‐induced MDA‐MB‐231 cells in vitro. Our data showed that TGFβ1‐induced MDA‐MB‐231 cell morphology change, promoted MDA‐MB 231 invasion, increased Vimentin expression, and decreased E‐cadherin expression. Further, Fangjihuangqi Decoction‐medicated serum (FHS) treated both MDA‐MB 231 cells and TGFβ1‐induced MDA‐MB‐231 cells. Results showed that Fangjihuangqi Decoction could inhibit cell proliferation, reduce cell invasion, increase E‐cadherin expression, and decrease EMT markers. Secondly, we established a xenograft tumor zebrafish model to assess Fangjihuangqi Decoction inhibition of cancer cell proliferation and invasion. Our results indicated that Fangjihuangqi Decoction could inhibit tumor growth, restrain the sprouts number of tumor neovascularization, and reduce the length of tumor neoplastic lymphatics by increasing E‐cadherin expression and decreasing EMT markers in TNBC xenograft tumor zebrafish model. Overall, our studies provide evidences that Fangjihuangqi Decoction could inhibit TNBC, reverse EMT, and contribute to antimetastasis by increasing E‐cadherin expression and decreasing EMT markers, which provide an experimental basis for clinical application of Fangjihuangqi Decoction on TNBC treatment.

Targets and Mechanism Used by Cinnamaldehyde, the Main Active Ingredient in Cinnamon, in the Treatment of Breast Cancer

“…The cell invasion assay was performed using Corning Transwell Chamber and Matrigel according to a previously described method ( Guo et al, 2020 ). Briefly, a serum-free medium was used to hydrate the Matrigel.…”

Modeling Cancer Using Zebrafish Xenografts: Drawbacks for Mimicking the Human Microenvironment

“…In other words, temperature and the time the embryos are exposed to a temperature greater than 28 °C needs to be evaluated and considered together. Various ranges of incubation times with different temperatures have been described in the literature [ 58 , 59 , 60 , 61 ], but to date no consensus has been reached. The latter is primarily due to the balance that needs to be struck between not only the temperature of the two components of the experiment (zebrafish embryos and human cells) but the temperature versus the incubation time.…”

“…To address this dilemma in the best possible way, most published studies have followed the standard protocol of 34°C and 6 days of incubation as previously described [ 39 , 52 , 53 , 54 , 55 , 56 ]. Recently, some studies have specifically modified this ‘standard’ protocol to try and customize it to different experimental settings, depending on what is being assayed or evaluated [ 59 , 60 , 65 ]. These studies highlight that xenotransplantation techniques cannot be forced to follow a standard protocol, instead, all experimental conditions, including the site of injection, temperature, and incubation time of the injected cells should be tailored to the experimental question and readout, in order to ensure that the most reliable and accurate in vivo results are obtained.…”

“…While standard protocols establish the normal incubation temperature at 34 °C, some studies aim to decrease the embryo incubation temperature when performing xenograft assays to 28–33 °C and the time down to 3–4 dpi [ 62 , 63 , 64 , 65 , 66 , 67 ], increasing the difference in terms of incubation temperature compared to the temperature of the human body, not allowing the cells to perform in their optimal conditions. On the other hand, other studies raise the incubation temperature, increasing the temperature to 35 °C, up from 34 °C [ 59 , 61 , 68 , 69 ], or even 36 °C [ 58 ], following the suggestion of some authors. It would be interesting to check what happens to the cell proliferation when the temperature is raised [ 20 ].…”

“…Hence, this information is not directly transferable to embryos incubated at 34 °C or more from 48 hpf after hatching, to 3–6 dpi. Reviewing the xenograft literature using zebrafish embryos, even when fish were incubated at 35 °C [ 59 , 61 , 68 , 69 ] the malformations highlighted by Pype et al [ 57 ], were not visible in imaged embryos. Apart from the different initiation and end time point conditions between the studies, and excluding the most critical developmental stage of the embryos, the lack of reported or documented malformation incidence could be due to the selection of the embryos without malformations when the first images of injected cells in the xenograft assays were performed.…”