An In Vitro Study: Overexpression Of Cystatin M On Invasiveness And Metastasis Of Breast Cancer Cell Line

Objective The present study aims to investigate the relationship betweenCystatin M and breast cancer and the effects of overexpression of Cystatin M on theproliferation,adherence,invasion and metastasis of breast cancer cell line.Methods Cystatin M gene was amplified by nPCR(nested polymearse chainreaction).Then the gene is cloned to pBudCE4.1 and amplified in E.coli strainstop10.Liposome was used to transiently transfer Cystatin M into ZR-75-30 cells,ahighly invasive human breast cancer cell line.The proliferation of ZR-75-30s andtheir adhesive ability to matrigel and endoepithelial cells was detected by MTTassays.The effects of Cystatin M on invasion and motility of the transfected cellswere investigated by reconstituted matrigel invasion and polycarbonate filtersmigration experiments.Results Overexpression of Cystatin M obviously reduced the adhesion ofZR-75-30 cells to matrigel and ECV304 cells(P＜0.01), and significantly retarded theinvasion and motility of the cells in matrigel-coated invasionchambers(P＜0.01),when compared with cells of control and parental group.However,the proliferation of ZR-75-30 cells was a little inhibited,and with no significance.Conclusion Our results showed that the adhesive and invasive ability of the humanbreast cancer cell line can be suppressed by Cystatin M.And it implies thatCystatin M may be a tumor metastasis suppressor gene and a new candidate for genetherapy against human breast cancer.