Effect And Mechanism Of Camptothecin Analogues FL118 On Invasion And Metastasis Of Human Breast Cancer

Objective To investigate the effect and mechanism of camptothecin analogues FL118 on invasion and metastasis of human breast cancer MDA-MB-231 cells.Methods Use high metastasis of human breast cancer MDA-MB-231 cells as the research object, to observe the effect of FL118 on tumor cell activity and proliferation by cell proliferation assay(MTT).Flow cytometry was used to determine the cycle distribution of tumor cell. Plate clonality formation assay, Plate wound healing assay and Transwell invasion assay were used to detect the change of the ability of plate clonality formation,transfer and invasive in human tumor cell after intervention by FL118.The m RNA expression of survivin, E-Cadherin and Vimentin which as signaling molecules were associated with Epithelial mesenchymal transition(EMT),β-catenin and Cyclin D1 which as the key nuclear protein were associated with Wnt signaling pathway were assayed by Real-time fluorescent quantitative PCR(q PCR).The protein expression of all of the above were detected by immunocytochemistry(ICC) and Western Blot hybridization(Western Blot).Results FL118 could inhibit the proliferation of MDA-MB-231 cells significantly in a dose-and time-dependent relationship(P<0.05), and the effect was better than that of the positive control group(P<0.05). FL118 causes MDA-MB-231 cells′mitosis to be blocked in the S phase(P<0.05). FL118 can decrease the plate colony formation ability of MDA-MB-231 cells(P<0.05).The results of Wound healing assay and Transwell invasion assay showed that FL118 could decrease the metastasis and invasion ability of MDA-MB-231 cells(P<0.05).The results of q PCR, ICC and Western Blot showed that FL118 could increased the m RNA and protein expression levels of E-Cadherin in MDA-MB-231 cells after intervention by FL118(P<0.05).The results of detecting by q PCR, ICC and Western Blot showed that these m RNA and protein expression levels including Vimentin、Survivin、β-catenin and Cyclin Dl were decreased in human tumor cell after intervention by FL118(P<0.05).Conclusion FL118 can inhibit the proliferation of breast cancer MDA-MB-231 cells, the plate colony formation ability, the metastasis and invasion ability, and can increase the expression of E-Cadherin, can decrease the expression of Vimentin and Survivin, can reduce the expression of nuclear β-catenin and Cyclin Dl.Preliminary study demonstrated that the mechanism of FL118 inhibition of the EMT is associated with Wnt signaling pathway.