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Objective To detect the effects of miR-143 on proliferation, migration and invasion of human nasopharyngeal cancer CNE-2Z cells.Methods The expression of miR-143 in NP69 cells, CNE-1 cells and CNE-2Z cells were detected by using real-time PCR.The overexpression of miR-143 in CNE-2Z cells was constructed through infecting lentivrius, and the level of miR-143 was determined by using real-time PCR.Cell viability was detected by using a CCK-8 kit according to the manufacturer's instruction at indicated time points.The effectiveness of overex-pression of miR-143 on migration and invasion of CNE-2Z cells were detected by using Transwell cell migration and invasion assay.Results The expression level of miR-143 in CNE-2Z was significantly lower than those in NP69 and CNE-1 (P<0.01).The miR-143 over-expressed CNE-2Z cell was successfully established.The cell viability in CNE-2Z/ miR-143 group was significantly decreased compared with CNE-2Z and CNE-2Z/miR-NC (P<0.01).Overexpression of miR-143 inhibited cell migration and invasion of CNE-2Z cells significantly(P<0.01).Conclusion miR-143 might inhibit cell proliferation, migration and invasion in human nasopharyngeal cancer CNE-2Z cells, indicating its important role in diagnosis, treatment and prognosis of nasopharyngeal cancer.
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Objective Taking CNE-2Z multicellular spheroids (MCSs) as the simulation of solid tumors, to investigate the role of SAPK/JNK signaling pathway in multicellular resistance to radiotherapy for human nasopharyngeal carcinoma (NPC). Methods Human NPC cell line CNE-2Z were cultured into multicellular spheroids by using liquid overlay technique, then divided into control MCSs, irradiated MCSs (average dose in one minute: 2 Gy), sp-600125(a specific inhibitor for SAPK/JNK signaling pathway)+irradiated MCSs, sp-600125+MCSs. Western blotting was employed to analyze the activity of SAPK/JNK signaling pathway in MCSs, and the expression of Caspase-3 protein before and after sp-600125 treatment; X-ray induced cell apoptotisis in MCSs before and after sp-600125 treatment was detected by TUNEL. Results The level of SAPK/JNK phosphorylation in MCSs was a dynamic course after radiation, and the phosphorylation peaked at 2 h after irradiation; The apoptotic rate of MCSs (P
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The effects of antisense oligonucleotides of c-fos and c-jun on the proliferation of CNE-2Z cells and on the expression of protein kinase C-? (PKC-?) were investigated. With the interpoising of Lipofectin(LP), the CNE-2Z cells were added with antisens oligonucleotides of c-fos and c-jun. MTT method was used to test the cell proliferation and flow cytometry (FCM) to the PKC-a expression. The results showed that the inhibition of antisense oligonucleotides of c-fos and c-jun to CEN-2Z cells was gradually enhanced with its concentration and prolonging time increasing and that its lowest effective concentration of LP was 1.7 X 10~(-6)?g/ml, the lowest effective concentration was of antisense oligonucleotides of c-fos and c-jun 1.7 x 10~(-5)?g/ml, and the most effective time was 20 hours after treatment. The fluoresence of PKC-a and percentage of positive cells in the groups treated by antisense oligonucleotides of c-fos and c-jun decreased significantly. The results indicated that the autisense oligonucleotides of c-fos and c-jun could inhibit the proliferation of CNE-2Z ceUs the expression of PKC-?.
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Objective:To investigate the mechanism of ursolic acid inducing the apoptosis of human nasopharyngeal carcinoma cell line CNE-2Z.Methods:CNE-2Z were treated by ursolic acid with different concentration.Cell proliferation was determined by MTT assay.Cell cycle and apoptosis rate were analysed by flow cytometry(FCM).The expressions of bax,bcl-2 and cox-2 were assessed by SP method of immunocytochemistry,while the expression of caspase 3 by Western blot.Results:Ursolic acid inhibited the proliferation of CNE-2Z cells and induced the apoptosis in dosage and time dependent manner.FCM showed that the cell population reduced in S phase and cell cycle was arrested in G0/G1 phase.Immunocytochemistry showed that the expression of bax was up-regulated in the apoptosis,while the expression of bcl-2 and cox-2 down-regulated.Western blot showed that the expression of caspase 3 was strengthened.Conclusion:Ursolic acid can induce apoptosis of CNE-2Z in vitro.Its molecular mechanism probably is through promoting activation of caspase3,up-regulating the expression of bax,and down-regulating the expressions of bcl-2 and cox-2.
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AIM To investigate whether tubeimosides induce apoptosis in CNE-2Z. METHODS Growth inhibition by tubeimosides was measured using MTT assay. The effect of tubeimosides on apoptosis induction of CNE-2Z cell line was studied by the fluorescent microscopy, electronic microscopy, DNA agarose gel electrophoresis, flow cytometry analysis. Western blotting was performed for detecting the apoptosis-related gene. RESULTS The growth of CNE-2Z cells was inhibited obviously by tubeimosides. CNE-2Z cells showed typical apoptotic features observed by fluorescent microscopy, electronic microscopy. DNA fragmentation into multiples of low molecular weight DNA(180～200 bp) was detected by agarose gel electrophoresis of DNA; Sub-G 1 peak was found by flow cytometry analysis, and the apoptosis index was 58 0%. The expression of bcl-2, inhibitor of apoptosis, decreased apparently; while that of bax, inducer of apoptosis, increased notably at 1, 3, 5 h after the addition of tubeimosides. The cleavage of caspase-3 was noticed at 1, 3, 5 h after the treatment of tubeimosides. CONCLUSION Tubeimosides could induce the apoptosis of CNE-2Z cells. The induction of apoptotic by tubeimosides was closely associated with bcl-2, bax and caspase-3.