The Effect Of Mouse Adipose Tissue-derived Stem Cells On Proliferation, Invasion And Metastasis Of Breast Cancer 4T1 Cells

Objective:To explore the effect of mouse adipose stem cells on the proliferation,invasion and metastasis of mouse breast tumor 4T1 cells through TGF-beta signaling pathway,providing a new idea for the clinical application of autologous fat filling.Methods:BALB/c mice were selected to obtain mouse adipose stem cells by isolation and culture of rat abdominal adipose tissue.The tumor cells were mouse breast tumor 4T1 cells.Divide the experiment into three groups: control group: 4T1 cells were cultured alone;Group 1:ADSCs were co-cultured with 4T1 cells;Group 2:neutralizing antibody against TGF-beta(Anti-TGF-beta)was additional added to the co-cultures of ADSCs with 4T1 cells.1?Flow cytometry and verification of the ability to induce differentiation was used to detect ADSCs;2?The proliferation of tumor cells was detected by CCK,RT-PCR;3?Tumor cell invasion and metastasis were detected by cell scratch test and Transwell cell migration test.Results:1?The isolated ADSCs showed fusiform fiber-like adherent growth,and showed differentiation into adipose and osteoblastic cells after 4 weeks of induction;2?Flow cytometry showed that the expressions of CD29 and CD44 were positive,while the expressions of CD45 and CD34 were negative;3?CCK method showed that compared with the control group and the group 2,the group 1 had significant proliferation of 4T1 cells at 48 hours(P<0.05);4?RT-PCR test showed that compared with the control group,PCNA expression in group 1 was significantly increased(p<0.01).Compared with group 2,PCNA expression in group 1 was statistically significant(p<0.05).Compared with the control group and the group 2,the expression of caspase-3 in group 1 was significantly reduced(p<0.05);5?Cell scratch test showed that compared with group 2 and control group,the cell scratch area healed faster in group 1;6?Transwell cell migration experiment showed that the group 1 had significant migration and invasion compared with the control group and group 2(P <0.01).Conclusion:1?ADSCs were successfully obtained from mouse abdominal adipose tissue;2?Mouse ADSCs can promote the proliferation and invasion and metastasis of mouse breast tumor 4T1 cells;3?Mouse adipose stem cells may affect the proliferation and invasion and metastasis of mouse breast tumor 4T1 cells through TGF-? signaling pathway.