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Objective To investigate the effect of curcumin on proliferation and apoptosis of human nasopharyngeal carcinoma (NPC) cell line CNE-2.Methods The CNE-2 cells were treated by by different concentrations (0,10,20,40,60 μmol/L) of curcumin.The proliferation activity of CNE-2 cells was detected by MTT assay,the cell cycle and apoptosis rate of CNE-2 were detected by using the flow cytometry (FCM),and the apoptosis was observed by Hoechest33258 fluorescence staining.Results Curcumin could significantly inhibit the proliferation of CNE-2 cells,moreover which was increased with curcumin concentration increase,the inhibitory rate of CNE-2 cells showed an increasing trend (P＜0.05),the half inhibitory concentrations (IC50) of curcumin acting on CNE-2 cells at 24,48,72 h were (23.54 ± 0.36),(18.31 ± 0.42) and (8.56 ± 0.37) μmol/L respectively.Curcumin could significantly inhibit the proliferation effect of CNE-2 cells,showing the apparent concentration and time dependence.The FCM detection results showed that in treating CNE-2 cells by 0,10,20,40,60 μmol/L of curcumin,the apoptosis rate was increased with the curcumin concentration increase;the fluorescence staining results showed that CNE-2 cells without curcumin treatment were round or oval,the cell nucleis were uniform in size,chromatin distribution showed the homogeneous light blue fluorescence;after 24 h of 10 μmol/L curcumin treatment,the CNE-2 cell body was shrunk and cell nuclear chromatin was condensed,showing granular bright blue fluorescence;after 24 h of 20 μmol/L curcumin treatment,the cell body was shrunk,nuclear was condensed,chromatin was uneven,apoptotic bodies appeared,and even the nuclear fragmentation appeared;after 24 h of 40 μmol/L and 60 μmol/L curcumin treatment,the number of apoptotic cells was increased,a large number of nuclear fragmentation appeared.Conclusion Curcumin has a significant inhibitory effect on the proliferation of NPC cell line CNE-2,moreover promotes the apoptosis of CNE-2 cells.
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Objective:To explore the proliferation inhibition and radiosensitization of Biyanqing granule on nasopharyngeal carci -noma cell line CNE-2 in vitro.Methods:CNE-2 cells were cultured in vitro.The inhibition of Biyanqing granule on the proliferation of CNE-2 cell was evaluated by MTT assay .Radiosensitization was explored by clone formation assay , and cell cycle and apopotosis were observed by flow cytometry ( FCM) .Results:Biyanqing granule could inhibit the proliferation of CNE-2 in a time-and dose-dependent manner.The IC50 in 24, 48 and 72 h was 70.79, 60.13 and 51.63 mg· ml-1(calculated according to the weight of all medicinal ma-terial), respectively.The colony formation assay showed that Biyanqing granule combined with radiation could significantly reduce the colony formation of CNE-2 cells.With the concentration increase of the main drug , the colony formation of CNE-2 cells was reduced . The number of colony formation in the negative control group , the radiation group , 10 mg· ml-1 and 20 mg· ml-1 Biyanqing combined with radiation groups (calculated according to the weight of all medicinal material ) was significant different (P<0.05).With the main drug concentration increasing , the percentage of G 2/M phase and apoptotic cells were both increased , and compared with the con-trol group, the difference was significant (P<0.05).Conclusion:Biyanqing granule can not only inhibit CNE-2 cells but also block CNE-2 cells in G2/M to improve the radiosensitization of CNE-2 cells.
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Background and purpose:Diallyl disulfide (DADS) is an oil-soluble organosulfur compound found in garlic. It can induce apoptosis in various cancer cell lines in vitro. This study investigated the effect of apoptosis of nasopharyngeal cancer cells line CNE2 induced by diallyl disulfide (DADS) and its molecular mechanisms. Methods:The growth inhibition and apoptosis of CNE2 cells and the protein levels of Bcl-2, Bax and Caspase 3 were examined by MTT assay, light microscopy, flow cytometry, DNA agarose gel electrophoresis and Western blot, respectively.Results:MTT assay showed that 50,100,200,400 ?mol/L DADS significantly inhibited proliferation of CNE2 cells at 48 hr, its inhibition ratio were 3.66%, 14.28%, 29.66%, 61.28%, respectively, in dose-dependent manner (P
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AIM:To explore the effects of small interfering RNA(siRNA) targeting PCNA gene on nasopharyngeal carcinoma CNE2 cells growth and cycle.METHODS:Three synthesized siRNA targeting PCNA gene was transfected into CNE2 cells by using LipofectamineTM reagent.The PCNA mRNA and PCNA protein were detected by real-time polymerase chain reaction(RT-PCR) and immunohistochemical method.Inverted phase contrast microscope was used to determine the CEN2 cells growth before and after PCNA-siRNA transfected.Flow cytometry was used to observe the cell cycle.RESULTS:In CNE2 cells after PCNA-siRNA transfection,the expressions of PCNA mRNA and protein were down-regulated at different degree.Inhibition ratio of PCNA mRNA was 98.5%.Meanwhile,the cell cycle was suffocated at G0/G1 stage.CONCLUSION:The synthesized PCNA-siRNA effectively interferes nasopharyngeal carcinoma cells by down-regulating the expressions of the PCNA mRNA and its protein,therefore inhibits the growth of CNE2 cells.Future application of PCNA-siRNA in the gene therapy of nasopharyngeal carcinoma might be expected.
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AIM: To discuss the discrepancy of microRNA (miRNA) in nasopharyngeal carcinoma (NPC) cells CNE-1 and CNE-2 on the basis of validating their different radioresistance.METHODS: Following the effect of X ray on the clones of CNE-1 and CNE-2 cells, the dose-survival curve and biological characteristics of CNE-1 and CNE-2 cells were determined by SigmaPlot software and the linear quadratic model of survival curves analysis. MicroRNAs were detected by Paraflo microfluidic microRNA chip, hybridization images collected using a laser scanner and normalizing the signals using a LOWESS filter. The relationship between the discrepancy of NPC radiosensitivity and the expression of microRNA was predicted according to Targetscan3.1 database (http:www.targetscan.org) after analyzing the data.RESULTS: Compared to CNE-2 cells, 20 microRNAs were gain-of-function and 13 microRNAs loss-of-function in CNE-1 cells among 326 detected microRNAs. 4 miRNAs that one detective value was more than 2000 and 3 folds than the other were hsa-miR-152, hsa-miR-7, hsa-miR-205 and hsa-miR-572. Data showed that radio-sensitivity was relative to the distinct discrepancy of miRNAs. CONCLUSION: The discrepancy of miRNAs is presents in different radioresistant NPC cell lines and related to radiosensitivity.
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Objective To establish a gemcitabine-resistant human nasopharyngeal carcinoma(NPC)cell line and study its characteristics.Methods The drug-resistant cell line CNE2/Gem was established by a procedure of treating the human NPC cells CNE2 with gemcitabine by pulse drug selection and continuous stepwise selection.The IC50 and resistance index(RI)to several commonly used anti-tumor drugs were tested by MTT assay.Fluorescence activated cell analysis(FACS)was employed for determining the cell cycle and concentration of fluorescence dye rhodamine 123 within the cells.Cell growth curve,doubling time and cell morphology were measured and observed.Results Duplication time of CNE2 and CNE2/Gem was 17.13h and 23.13h,respectively,as evaluated by the growth curve.The IC50 to gemcitabine increased from 2.84?1.63?g/ml in CNE2 to 42.95?7.53?g/ml in CNE2/Gem as tested by MTT assay at 48～72h exposure,the RI was 23.61(P