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OBJECTIVE：To study the inhibi tory effects of genistein on the growth of human nasopharyngeal carcinoma. CNE 1 cells and predict its potential target. METHODS ：CCK-8 method was used to test the effects of 0（blank control ），12.5，25，50， 100，150 µmol/L genistein on the proliferation of CNE 1 cells after treated for 24，48，72 h. Flow cytometry was carried out to detect the effects of 0（blank control ），15，30，60 µmol/L genistein on the cell cycle and ap optosis of CNE 1 cells after treated for 24 h. Scratch test was used to investigate the effects of 0（blank control ）， 10， 20， 30 µmol/L genistein on themigration ability of CNE 1 cells after treated for 24 h. High （No.18210156） throughput sequencing was conducted to discover the differential genes in CNE 1 cells after treated with 0（blankcontrol），30 µmol/L genistein for 24 h. RT-qPCR assay was adopted to verify the mRNA expression of related differential genes in above trials. RESULTS ： Compared with blank control，12.5，25，50，100，150 µmol/L genistein sho wed significant inhibitory effect on the proliferation of CNE 1 cells（P＜ 0.01），in a concentration- time-effect manner ；15，30 µmol/L genistein could arrest CNE 1 cell cycle at G 0/G1 stage（P＜0.05 or P＜ 0.01）；30，60 µmol/L could arrest CNE 1 cell cycle at G 2/M stage and promoted cell apoptosis （P＜0.05 or P＜0.01）. 10，20，30 µmol/L genistein could significantly inhibit the migration ability of CNE 1 cells（padj＜0.01）. High throughput sequencing revealed a total of 2 271 differentialgenes（P＜0.05），1 154 of which were up-regulated while 1 117 of which were down-regulated ；8 potential target genes ，including p53，p21，STC2，FGF2，CDK6，CYCLIN D ，PI3K，AKT，were screened by cell experiment. After validated by RT-qPCR assay ，mRNA expression of p53，p21，STC2，FGF2，CDK6，CYCLIN D and AKT were significantly down-regulated（P＜0.05），which consistent with the sequencing results. CONCLUSIONS ：Genistein can effectively inhibit the growth of human nasopharyngeal carcinoma CNE 1 cells，the mechanism of which may associated with inhibiting the expression of mutant gene p53，restoring the function of wild-type P 53 protein and inhibiting the activity of PI 3K/Akt pathway.
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objective To research whether the combination of chrysin and camptothecin can promote the apoptosis of human nasopharyngeal carcinoma cell line CNE2 and to explore the molecular mechanism of the combinative effect. Methods: CNE2 cells were pretreated with designed dose of chrysin (10|, 20, and 40 μmol/L) for 2 h, then treated with camptothecin (1 μg/mL) for 24 h. The morphologic changes were observed under inversed microscope and the cell viability was measured using MTT. The activity of caspase-3 and PARP, which was regarded as the protein marks of apoptosis, was determined by Western blotting. Then the cells were treated with chrysin for different time and the time course of apoptosis inhibitory protein, Bcl-xL was also detected using Western blotting. Results: Increases of cell death were observed in the group with combined chrysin and camptothecin, but no obvious cell death could be found in chrysin, camptothecin alone, and control groups; The data of cell viability supported this results; With the enhance of pretreatment dose of chrysin, the cell viability decreased. There were the significant differences between the combined groups and the control one (P<0.05), and between the combined groups and both the chrysin and camptothecin groups separately (P<0.05). Chromatin condensation, which was the indication of apoptosis, could be observed when the cells were stained with Hochest 33342; The proprotein of caspase-3 and PARP degraded and there were the dose-dependent and time-dependent effect. The pan-caspase inhibitor Z-VAD-fmk could inhibit the apoptosis of CNE2 cells which were treated with the combination of chrysin and camptothecin, according to the cell viability and the activation of caspase-3 and PARP; The time-dependent down-regulation in the apoptosis inhibitory protein Bcl-xL could be observed. Conclusion: The cotreatment of chrysin and camptothecin could promote the apoptosis of CNE2 and the down-regulation of apoptosis inhibitory protein Bcl-xL played an important role in the combinative effect.
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Objective:To investigate the inhibitory effect and its possible molecular mechanisms of MicroRNA-34a(miR-34a) on the human nasopharyngeal carcinoma CNE-2 cell line subcutaneous xenograft tumor in nude mice.Methods: The human nasopharyngeal carcinoma CNE-2 cell line was cultured in vitro.miR-34a and Scrambled miRNA recombinant plasmids were successfully established and stably transfected into CNE-2 cells.Fifteen six-week-old male nude mice were divided randomly into three groups:miR-34a group(5 mice) ,Scrambled miRNA group(5 mice) ,Blank control group(5 mice).Different CNE-2 cells were subcuta-neously injected on the back near right lower limb.Tumor volumes were examined every 7 days.Mice were executed on the 35 days,and the eventual average tumor volumes and weights were examined.Total RNA and protein were isolated from tumors,and the expression of miR-34a,CDK6,and Bcl-2 mRNA and protein were determined by qRT-PCR and western blot,respectively.Results: The relative expressions of miR-34a was significantly up-regulated in miR-34a transfected group compared to Scrambled miRNA transfected group (P (849.62±101.32) mm3 ,respectively,and the eventual average tumor weights in miR-34a group,Scrambled miRNA group and blank control group were(0.81±0.13)g,(1.47±0.21)g and(1.58±0.37)g,respectively.Both the eventual average tumor volumes and weights in miR-34a group were lower compared to the other two groups(P<0.05).qRT-PCR results revealed that the expression of miR-34a in miR-34a transfected group was significantly higher than in the other two groups,while the mRNA and protein expression of CDK6 and Bcl-2 were lower than the other two groups ( P<0.05 ) .Conclusion: miR-34a may inhibit the growth of human nasopharyngeal carcinoma CNE-2 cell line subcutaneous xenograft tumor in nude mice by down-regulating CDK6 and Bcl-2.
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0.05).The mRNA expression of Cyclin B1 of IMRT group was significantly higher than that of ART group at each dose point(P
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Background and purpose:Somatostatin receptors have been found in a variety of tumors and are therefore amenable to treatment with somatostatin analogs, like octreotide. However, the study of SSTRs expression has been rarely studied in nasopharyngeal cancer.We investigated the expression of somatostatin receptors gene subtypes in human nasopharyngeal cancer cell line CNE_ 2 . Methods:We have harvested cultured human nasopharyngeal cancer cell line CNE_ 2 . Using both techniques, reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemical assay, we analysed mRNA of different subtypes of somatostatin receptors in human nasopharyngeal cancer cell line CNE_ 2 .Results:The positive rate of somatostatin receptors subtype SSTR_ 1 SSTR_ 2 SSTR_ 4 was manifested in the human nasopharyngeal carcinoma cell line CNE_ 2 by RT-PCR and immunohistochemical assay. According to immunohistochemical assay, SSTR_ 1 and SSTR_ 2A showed strongly positive expression and SSTR_ 3 and SSTR_ 5 negative expression,respectively.Conclusions:There are more than one SSTR subtypes expressed in the human nasopharyndeal carcinoma cell line CNE_ 2 . This study demonstrated the presence of SSTR_1,SSTR_ 2 in the human nasopharyndeal carcinoma cell line CNE_ 2 .
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OBJECTIVE:To evaluate the inhibitory actions of 3 traditional Chinese drugs on human nasopharyngeal carcinoma cells CNE-2 in vitro.METHODS:The IC50(50% inhibiting concentration)of 3 traditional Chinese drugs on human nasopharyngeal carcinoma cells CNE-2 in vitro was measured by MTT assay.RESULTS:The inhibitory actions of 3 traditional Chinese drugs on human nasopharyngeal carcinoma cells CNE-2 in vitro were enhanced with the increase of the concentration in a concentration-dependent manner,with formulation Ⅲ showing the most potent inhibitory action on CNE-2 cells in vitro.CONCLUSION:The heat-clearing and detoxicating traditional Chinese drugs could markedly inhibit the proliferation of CNE-2 cells.
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Objective:To investigate the effects of tetrandrine(Tet) on proliferaton and apoptosis of nasopharyngeal carcinoma cell line CNE.Methods:CNE cells were divided into groups and treated by Tet at different concentrations.Inhibitory effects of tetrandrine were determined by MTT assay.The morphologic changes of CNE cells were observed by transmission electron microscope.DNA fragmentations were determined by gel electrophoresis assay.Cell apoptosis was investigated by flow cytometer.The expressions of apoptosis-related genes were detected by retrotranscriptase polymerase chain reaction(RT-PCR).Results:Tetrandrine possessed inhibitory effect on CNE cell proliferation in a concentration-dependent manner(P
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Objective To study the inhibitory effect of berberine on human nasopharyngeal cancer cell line CNE-2 in vivo and in vitro. Methods CNE-2 cell proliferation was measured by MTT assay and cell cycle was analyzed by flow cytometry. Cell morphology was observed with transmission electron microscopy. The cell cycle relative protein was detected by Western blotting. DNA and protein syntheses were assessed by the cellular incorporation of ~ 3 H-TdR and ~ 3 H-Leu, respectively. Anti-tumor activity of berberine in the experimental transplantation tumor CNE-2 was evaluated by relative tumor growth ratio. Results Berberine inhibited CEN-2 cells growth in a time-and dose-dependent manner. MTT Assay showed that the IC_ 50 values of 48 and 72 h were (49.5?5.8) and (13.3?2.0) ?mol/L, respectively. Cell cycle analyses of 50.0 ?mol/L berberine-treated CNE-2 cells by flow cytometry showed the accumulation of cells in the G_2/M phase while 25.0 ?mol/L berberine treatments for 48 h induced apoptosis with the index of (48.9?10.4)%. The inhibition of CNE-2 cell growth by berberine was associated with suppression of cyclin B1, CDK1, and cdc25c proteins. After the treatment of berberine at dose of 30 mg/kg, the median tumor volume was 317.9 mm~3 which was much lower than that in control group (P
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Actinomycin 23-21 ( ACT 23-21 ) is an anticancer antibiotic of Actinmycines. This antibiotic was produced from soil Streptomyces flaveolus which was isolated and obtained from soil samples in Fuzhou, China.The effects of ACT 23-21 were observed on using 2 transplanted models of human nasophryangeal carcinomatous cells ( CNE-2Z ) and human gastric carcinomatous cells ( MGc-803 ) in nude mice. At ACT 23-21 50?g/kg, the inhibition rate for transplanted tumors of CNE-2Z and MGc-803 were 58.4% (P