Carcinoma-associated Fibroblasts(CAFs) And Wnt Genes Promote The Proliferation Of Human Breast Cancer Cells

Objective: To explore the influencing role of Wnt genes derived from CAFs onhuman breast cancer cells through Wnt signaling pathway.Methods:1.6normal breast tissue specimens and6breast cancer tissue specimenswere obtained from6Primary breast cancer female sufferers. These patients had nopreoperative treatment and their pathological classifications are invasive breast cancer.Extract and culture CAFs and NBFs from these tissue specimens.2. Setting up Matrigel Three-Dimensional cell culture models: control group (CG)1(human breast cancer cells MCF-7only), CG2(MCF-7+NBFs), experimentalgroup (EG)1(MCF-7+CAFs), EG2(MCF-7+CAFS+anti-Wnt3a Ab), EG3(MCF-7+CAFS+anti-Wnt5a Ab), EG4(MCF-7+CAFS+anti-Wnt10b Ab). Repeat3times. Compare the rates of cell clone.3. Co-culture CAFs/NBFs and MCF-7. Compare the Wnt mRNA levels of theMCF-7by Real-time quantitative PCR.Result: In the Matrigel Three-Dimensional cell culture models: the rate of cells cloneof EG1is higher than CG1and CG2; the rates of cells clone of EG2and EG4arelower than EG1and CG2, but still higher than CG1and CG2; the rate of cells clone ofEG4is lower than EG2; the rate of cells clone of EG3is lower than EG1.The expression of Wnt3a, Wnt5a and Wnt10b gene in experimental groups aresignificantly higher than control group. The expression of Wnt3a and Wnt10b gene inEG2are significantly higher than EG1. The expression of Wnt5a in EG2issignificantly lower than EG1.Conclusions: The over-expression of Wnt3a and Wnt10b in the CAFs enhance the proliferation in human breast cancer cell lines. The function of Wnt5a is not clear.Wnt signals may influence the self-renewal of stem cells in mammary gland.