The Effect And Mechanism On The Migration Of HUVEC Migration By Mitomycin

Objective:The aim of the present research is to study the selectivity of Mitomycin on human umbilical vein endothelial cells (HUVEC) function and cell migration and the mechanism was also discussed.Methods:(1) Effect of Mitomycin on the selectivity of HUVEC. Comparing with the Human hepatoma SMMC-7721 cell (7721) as the control of different doses of mitomycin and cells were cultured, HUVEC's proliferation was measured by MTT and the different groups'half effective inhibitory concentration (IC50) was for the role of selective index to measure the selective effect of Mitomycin on HUVEC.(2) Mitomycin on HUVEC apoptosis. HUVEC are cultured with different doses of Mitomycin were for 48 hours. The cells are fixed and Cell morphology are assayed by HE staining and the micrography of the apoptosis cells was analyzed by AO:EB double staining.(3) Mitomycin on HUVEC migration. Detect changes in HUVEC imgration by Transwell small room (Boyden Chamber Transwell), Remove the upper small chamber and observate HUVEC migration under the micro-scope after HUVEC are cultured with different doses of Mitomycin were for 8 hours.(4) Mitomycin on HUVEC AQP1 expression. After cultivating HUVEC with different doses of MMC for 48 hours, detect the expression of AQP1 by Immunocytochemistry and Western-Blot method.Results:(1) Mitomycin has the selective inhibitory effect on HUVEC. Training 24,48,72 hour IC50 were 34.74μg/ml,1.79μg/ml,2.03μg/ml, and 7721 control cells (379.05μg/ml,11.12μg/ml,5.21μg/ml) difference was significant (P＜0.01).(2) Mitomycin on HUVEC apoptosis can promote. HE stainin showed that treatment with different doses of mitomycin HUVEC reduced the volume of varying degrees occurs, nuclear condensation, cell damage such as deep-stained morphological changes. AO:EB double staining, using different doses of Mitomycin for 48 hours, the experimental group and control group, the apoptosis rate increased with the dosage increased (P＜0.01).(3) Mitomycin on HUVEC migration was inhibited. Transwell small chamber (Boyden Chamber Transwell) obserbed the impact of Mitomycin on HUVEC migration, results showed that the dose does not affect cell viability compared with the normal control group, high dose (0.09μg/ml),medium dose (0.03μg/ml),low dose (0.01μg/ml) Mitomycin can decreased HUVEC migration rate decreased 75.1%,49.5%,18.4%.(4) Mitomycin inhibited HUVEC APQ1 protein expression. Immunocytochemistry determination of mitomycin treatment on HUVEC AQP1 expression, the results show that the drug group compared with normal cells in culture cells were found to reduce the number of AQP1 expression, and there is a dose dependent manner (P＜0.01).Conclusion:1. Role in mitomycin significantly inhibited HUVEC proliferation in a dose-dependent, and high selectivity compared to control cells,7721.2. HUVEC apoptosis induced by mitomycin inhibited HUVEC prolife-ration.3. Mitomycin inhibited cell migration when does not affect HUVEC survival.4. HUVEC AQP1 expression may be induced with mitomycin apoptosis, anti-migration related to the specific mechanism remains to be further studied.