| Extensive research and experiments confirmed that 5-FU nanoparticles can inhibit the proliferation of Gastrointestinal carcinoma; but few study of the molecular mechanism of the nanoparticles. To this study,5-FU-PLA-O-CMC-NPS may inhibit t SW480 colon cancer cell proliferation through the G1/S cell cycle arrest which related the regulatory factors P21WAF/CIP, CyclinD1and Rbo.MethodsLA tetrazolium (MTT) assay the inhibition rate of SW480 affected by 5-FU-PLA-O-CMC-NPS from,24,48,72120h with various concentrations including0.1ug/ml,1 ug/ml,10 ug/ml and 100 ug/ml on the value-added.And observe 5-FU-PLA-O-CMC-NPS controlled-release characteristics compare with 5-FU.2.Flow cytometry (FCM) detection 0,24,72,120 h with different concentrations including 0.1ug/ml,1 ug/ml,10 ug/ml and 100 ug/ml 5-FU-PLA-O-CMC-NPS on SW480 cell cycle, and further clarify the inhibit the proliferation mechanism of colon cancer cell.3.Westernblot and RT-PCR using, from the protein and mRNA levels to study the proliferation inhibition on SW480 and obseraving the expression changes of the key factors P21WAF/CIP, CyclinD1, Rbwhich lead to G1/S cell cycle arrest.Results1.MTT results showed that 5-FU-PLA-O-CMC-NPS on human colon cancer cell line SW480 was a dos-time-dependent compared with 5-FU, 5-FU-PLA-O-CMC-NPS has a good release effect. There wao no significant difference between.5-FU-PLA-O-CMC-NPS and 5-FU (P> 0.05).2.FCM results showed that different concentrations of 5-FU-PLA-O-CMC-NPS (0.1ug/ml,1 ug/ml,10 ug/ml,100 ug/ml) effecting on SW480 for72h mainly leading to G1/S phase arres.G1 has a significant increasing in the proportion of 58.74%±1.22 increased to 81.33%±2.94 with significant difference (t=-24.027, P = 0.032); S phase increased from 37.37%±3.42 decreased to 14.82%±3.41, with significant difference (t= 6.098, P= 0.015). 10μg/ml 5-FU-PLA-O-CMC-NPS role in SW480 cells, respectively 24h,72h,120h and the drug did not increasing compared with the control group.Gl phase was increased:from 70.66%±0.90 increased to 78.32%±0.14 (t=-6.504, P= 0.022).S phase were decreased:from 28.43%±2.75 decreased to 18.66%±1.42 (t= 4.296, P= 0.016) and G2/M phase was no significant change (P> 0.05).3.Westernblot and RT-PCR results showed that:P21WAF/CIP, cyclinD1, Rb expression changes of SW480 on 5-FU-PLA-O-CMC-NPS shows a dose-time dependent..P21WAF/CIP, Rb mRNA and protein expression increased proportional with 5-FU-PLA-O-CMC-NPS concentration; cyclinDl mRNA and protein expression decreased i with P21WAF/CIP.Conclusions1 The preparation and release of the 5-FU-PLA-O-CMC-NPS had no effect on inhibiting tumor effect, and compared, SW480 cell proliferation inhibition of the slow onset and long duration with 5-FU.2.5-FU-PLA-O-CMC-NPS induced proliferation inhibition for SW480 cells through the G1/S cell cycle arrest.3.5-FU-PLA-O-CMC-NPS inhibition mechanism independent on wild-type P53 way.It was induced by the key of the regulatory factor P21WAF/CIP, while playing down Cyclin family and increase Rbo to arrestin G1/S cell cycle phase...
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