Effect And Mechanism Of Niclosamide On Improving The Apoptosis Of SKOV3 Ovarian Cancer Cell Lines

Objective:To observe the effects of anti-helminthic drug named niclosamide on promoting cell’s apoptosis in ovarian cancer cell line SKOV3, and its mechanisms of the drug targets on Wnt/β-catenin pathways.Methods:1. The cytotoxicity of various doses of niclosamide on SKOV3 was detected by MTT assay, and drew the moderate doses(4umol/l and 8umol/l) to continue the following tests.2. The cytotoxicity of niclosamide combined with DDP on SKOV3 was also detected by MTT assay.3. The flow cytometry was taken to find out the function of niclosamide and niclosamide combined with DDP in improving apoptosis of SKOV3 cells.4. Cells were treated with niclosamide(4um/l, 8um/l) ±DDP(10um/l), then m RNA and protein levels of β-catenin and CyclinD1 were investigated in SKOV3 cells by RT-PCR and Western Blot.5. Analy the experiment’s data.Results:1.MTT assay results revealed that niclosamide can inhibit the proliferation of SKOV3 cells in a dose-time-dependent manner. 50% of SKOV3 cells will be suppressed when treated with 6.29umol/l niclosamide for 24 hours(IC50). The same effect will be gotten with 3.55umol/l niclosamide for 48 hours and 1.31umol/l niclosamide for 72 hours. Then we choose 4umol/l and 8umol/l niclosamide to continue our experiment.2. MTT assay results revealed that niclosamide at 8umol/l inhibits SKOV3 cells more significantly than niclosamide at 4umol/l(P=0.000). The inhibition of niclosamide with DDP shows no difference with that treated niclosamide alone(P>0.05).3. The flow cytometry showed that there is no difference between niclosamide with DDP group and niclosamide alone group in the effect of promoting apoptosis.4. The results of PCR and Western Blot showed that when SKOV3 cells treated with niclosamide for 24 hours, the mRNA and protein levels of β-catenin and CyclinD1 are significantly lower than blank group. The mRNA and protein levels ofβ-catenin and CyclinD1 decrease when the dose of niclosamide goes up(P=0.000).The levels measured from SKOV3 cells treated with niclosamide with DDP for 24 hours have no significant difference with the group SKOV3 cells treated with niclosamide alone(P>0.05).Conclusions:1. Anti-helminthic drug niclosamide inhibit the growth of SKOV3 cells and have a tight correlation with time and dose.2. Niclosamide inhibits Wnt/β-catenin signaling pathway,and the inhibition effect would be more obvious with a higher dose of niclosamide.3. Niclosamide promotes SKOV3 cells’ apoptosis probably through reducing the mRNA and protein levels of β-catenin and CyclinD1 in Wnt/β-catenin signaling pathway.4. Niclosamide and DDP didn’t show some synergetic effects in inhibiting SKOV3cells’ proliferation.5. DDP playing the role of inducing apoptosis of SKOV3 cells may have little relationship with Wnt/β-catenin signaling pathway.