c-Kit-Positive Adipose Tissue-Derived Mesenchymal Stem Cells Promote the Growth and Angiogenesis of Breast Cancer

Figure 2

Effects of c-Kit expression of ASCs on 4T1 breast cancer cells. ASCs and 4T1 cells were cocultured in a ratio of 1 : 1. After culturing, RNA and protein were extracted to detect the c-Kit mRNA and protein expression in different culture models. After 1–5 days, the viability and proliferation of 4T1 cells were detected using a CCK-8 assay and DNA quantification, respectively, in indirect coculture experiments. For the tube formation assay, 10⁴ ASCs and 10⁴ EPCs were plated on Matrigel, respectively, and incubated for 18 h, and then the area of tube formation was observed. (a) The mRNA expression of c-Kit was higher in the c-Kit⁺ASCs + 4T1 direct coculture group than in the single culture groups using qPCR, and the expression of c-Kit mRNA was not detected in the 4T1 cells. The qPCR results were normalized against GAPDH. (b) The level of c-Kit protein expression in the c-Kit⁺ASCs + 4T1 direct coculture group was higher than that in the single culture groups by western blot analysis. But no c-Kit protein expression was detected in the c-Kit^- ASCs + 4T1 group. Anti-β-actin antibody served as a control. ((c)-(d)) The viability and proliferation of 4T1 cells were enhanced by c-Kit⁺ ASCs, compared to the other culture groups. (e) The c-Kit⁺/c-Kit^- ASCs had no potential of tube formation compared with EPCs. ; ; . (e): 200x magnification.

(a)

(b)

(c)

(d)

(e)