| Objective: To study the anti-tumor mechanisms and effect of curcumol on the human Nasopharyngeal Carcinoma (NPC) CNE-2 cells, and to explore its impacts on the expressions of Nuclearfactor Kappa B (NF-κB) and Anti-apoptosis Protein (Bcl-2). Both the general investigation of apoptosis-inducing effect of curcumol on CNE-2 cells and the preliminary exploration of molecular mechanisms involved in the present research tried to provide experimental support for the anti-tumor effect of curcumol in clinical application. Methods: Curcumol was dissolved in dehydrated alcohol, and initial concentration of curcumol was 10mg/mL. The curcumol concentrations were set as 12.5, 25, 50, 100μg/mL, and 1% dehydrated alcohol was taken as the control group. Four methods used in this thesis: MTT colorimetric assay was applied to detect the anti-proliferation effect of different concertration curcumol (12.5, 25, 50, 100μg/mL) for 24, 48, 72, 96 and 120h respectively on CNE-2 cells; Nuclear morphological changes of CNE-2 cells in apoptosis process treated by curcumol for 48hs were determined by Hoechst 33342 fluorescent staining; Flow Cytometer (FCM) was used to analyze the apoptotic rates of CNE-2 cells induced by curcumol for 48hs; The expressions of NF-κB and Bcl-2 on CNE-2 treated by curcumol for 48h were detected by Western Blot. Results: MTT colorimetric assay showed that proliferation of CNE-2 cells could be restrained by 25-100μg/ml curcumol for 48, 72, 96 and 120h respectively. With the increase of curcumol concentration and incubation time, the inhibition was reinforced. Specificly, compared with the control group, drug groups (25, 50, 100μg/mL) have significant decrease (p<0.05), while no obvious decrease (p>0.05) in drug group (12.5μg/mL). Through Hoechst 33342 fluorescent staining, the nuclear morphological changes of CNE-2 cells treated by 100μg/ml curcumol for 48h showed as condensation, aggregation, pyknosis and fragmentation under light microscope. Morever, under fluorescent microscope, both densely stained bright blue fluoresence of apoptosis nulear and apoptosis body could be seen in the CNE-2 cells treated by curcumol for 48h, while the normal cell nuclear in the control group presented with diffused uniform bluish fluoresence. Flow Cytometry analysis revealed that different apoptosis rates of CNE-2 cells treated by different concentration curcumol for 48h. Compared with the control group, drug groups (12.5, 25μg/mL) have no significant difference (P>0.05), while drug groups (50, 100μg/mL) have statistical sense (P<0.01) and characterized with obvious concentration-dependence. Western Blot analysis showed that 12.5-100μg/ml curcumol could decrease the expressions of NF-κB and Bcl-2. After cells treated by curcumol for 48 h, the expresstions of NF-κB and Bcl-2 were down-regulated with the continuous increase of curcumol concentration and incubation time. Compared with the control group, drug groups have significant decrease (P<0.05) and make relativity relationship between NF-κB and Bcl-2 (r=0.975, P<0.01). Conclusions: Curcumol could restrain proliferation of CNE-2 cells effectively after incubation of 24, 48, 72, 96 and 120h, the inhibition of cell proliefration might be performed as a time-dose dependent manner; curcumol could induce apoptosis of CNE-2 cells after incubation of 48h in vitro; NF-κB expression was inhibited after CNE-2 cells treated by curcumol for 48h, and this effect could be viewed as one possible anti-tumor mechanism by curcumol; To accelerate cells apoptosis, Bcl-2 expression down-regulated after CNE-2 cells treated by curcumol for 48h, and in addition, there is a correlation between Bcl-2 expression and NF-κB expression. |
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