Extracellular Microenvironmental Change by B16F10 Melanoma-derived Proteins Induces Cancer Stem-like Cell Properties from NIH3T3 Cells

Soon Yong Park,Hyeongrok Choi,Jeong-Ki Min,Jin Woong Chung,Sang-Kyu Ye,Sun-Hee Leem,Sangin Shim,Joo Eon Lee,Ae Jin Jeong,Dong Gwang Lee,Woojin Jun,Ara Jo

doi:10.1038/s41598-019-53326-8

Abstract

Cancer stem-like cells (CSCs) can generate solid tumors through the properties of stem cells such as self-renewal and differentiation and they cause drug resistance, metastasis and recurrence. Therefore, establishing CSC lines is necessary to conduct various studies such as on the identification of CSC origin and specific targeted therapies. In this study, we stimulated NIH3T3 fibroblasts to exhibit the characteristics of CSCs using the whole protein lysates of B16F10 melanoma cells. As a result, we induced colony formation that displayed self-renewal and differentiation capacities through anchorage-independent culture and re-attached culture. Moreover, colonies showed drug resistance by being maintained in the G0/G1 state. Colonies expressed various CSC markers and displayed high-level drug efflux capacity. Additionally, colonies clearly demonstrated tumorigenic ability by forming a solid tumor in vivo. These results show that proteins of cancer cells could transform normal cells into CSCs by increasing expression of CSC markers. This study argues the tremendous importance of the extracellular microenvironmental effect on the generation of CSCs. It also provides a simple experimental method for deriving CSCs that could be based on the development of targeted therapy techniques.

Highlights

Cancer stem-like cells (CSCs) were found in various solid tumors, such as bladder, breast, brain, colon, prostate, pancreatic cancer, and melanoma after the first CSCs had been found in acute myeloid leukemia in 19973
We found that the treatment with protein lysates of B16F10 melanoma cells could transform NIH3T3 cells into the colony form, which possessed the characteristics of CSCs
We observed a definite induction of colony formation by the B16F10 cell-derived proteins, while the boiled B16F10 cell-derived proteins could not induce colony formation in the NIH3T3 cells (Fig. 1B), suggesting that the major factors for the colony formation are the proteins in the cell lysates

Summary

Introduction

CSCs were found in various solid tumors, such as bladder, breast, brain, colon, prostate, pancreatic cancer, and melanoma after the first CSCs had been found in acute myeloid leukemia in 19973. CSCs are induced by the accumulation of gene mutations in normal stem cells This hypothesis can be the basis for demonstrating the differentiation potential of CSCs. Third, already differentiated cells undergo dedifferentiation and acquire similar characteristics to stem cells. Already differentiated cells undergo dedifferentiation and acquire similar characteristics to stem cells These stem cell-like cells undergo epithelial–mesenchymal transition, which is a key process in tumorigenesis, leading to CSC formation through the transformation of the stem cells[9,10]. This study aimed to determine whether the extracellular microenvironment change by various intracellular components of cancer cells could convert mouse fibroblasts into putative CSCs. Surprisingly, we found that the treatment with protein lysates of B16F10 melanoma cells could transform NIH3T3 cells into the colony form, which possessed the characteristics of CSCs

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