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Background Astragalus polysaccharide (APS) had shown great promise in anti-tumour activities in our previous studies. The present study was designed to investigate whether APS has synergistic effect with cisplatin on the growth-inhibitory of human nasopharyngeal carcinoma cell lines and the possible mechanism. Methods Here, nasopharyngeal carcinoma cell lines (CNE-1) were divided into CNE-1 group, Cisplatin treatment group (2 µg/mL Cisplatin), APS treatment group (200 µg/mL APS) and combination group (2 µg/mL Cisplatin and 200 µg/mL APS). The proliferation inhibition rate of CNE-1 cells was determined by Cell Counting Kit-8 (CCK-8) method after treatment with different concentrations of APS for 24, 48, and 72 h. Apoptosis rates and cell cycle retardation of cells were detected by flow cytometry. Cell migration and invasion was evaluated by transwell assay. Western blotting and quantitative (q)RT-PCR were performed to detect the expression of Bcl-2, Bax, caspase-3, matrix metalloproteinase-2 (MMP-2), p53 and matrix metalloproteinase-9 (MMP-9) proteins in CNE-1 cells. Results APS have an inhibition on the proliferation of CNE-1 cells with time and dose dependence manner. Both the APS and combination therapy could promote apoptosis of CNE-1 cells, with the count of cells increased in G0/G1 and S phase while decreased in G2/M phase, and inhibited the migration and invasion of CNE-1 cells. Moreover, co-administration of Cisplatin and APS was more efficacious for the antitumor effect than either agent alone, as evidenced by the significant decrease in MMP-9 level and increase in p53. Conclusion APS, in combination with cisplatin, had significantly synergistic growth-inhibitory effect on nasopharyngeal carcinoma cell lines, which may be related to cell cycle and migration induction.
Subject(s)
Cell Cycle/drug effects , Cell Movement/drug effects , Cisplatin/pharmacology , Nasopharyngeal Carcinoma/drug therapy , Polysaccharides/pharmacology , Apoptosis/drug effects , Caspase 3/metabolism , Cell Line, Tumor , Cell Proliferation/drug effects , Drug Combinations , Humans , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/metabolism , Proto-Oncogene Proteins c-bcl-2/metabolism , Tumor Suppressor Protein p53/metabolismABSTRACT
Objective To investigate the effect and underlying mechanism of lncRNA MEG3 on the radiosensitivity of nasopharyngeal carcinoma cells.Methods this experiment,overexpression control group,MEG3 overexpression group,miR-NC inhibition group,miR-7-5p inhibition group,overexpression control+4 Gy group,MEG3 overexpression+4 Gygroup,miR-NC inhibition+4 Gy group,miR-7-5p inhibition+4 Gy group,MEG3 overexpression + miR-NC overexpression group,MEG3 overexpression + miR-7-Sp overexpression group were established.The expression of miR-7-5p and MEG3 was detected by qRT-PCR.The radiosensitivity of nasopharyngeal carcinoma cells was measured by clone formation assay.Cell apoptosis was assessed by flow cytometry.The fluorescence activity was evaluated by dual luciferase reporter assay.Results MEG3 was lowly expressed in nasopharyngeal carcinoma tissues and cells.Overexpression of MEG3 and inhibition of miR-7-5p expression increased the radiosensitivity of nasopharyngeal carcinoma cells and promoted radiation-induced cell apoptosis.MEG3 could targetedly regulate the miR-7-5p expression.Overexpression of miR-7-5p reversed the effect of overexpression of MEG3 on the sensitization of nasopharyngeal carcinoma cells and the promotion of apoptosis induced by radiation exposure.Conclusions Overexpression of MEG3 increases the radiosensitivity of nasopharyngeal carcinoma cells and promotes radiation-induced cell apoptosis.The mechanism may be related to the down-regulation of miR-7-5p expression.
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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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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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Objective:To explore the effect of inhibiting mdr1 by antisense technology on radiosensitivity in nasopharyngeal carcinoma cell line(CNE). Methods:There were four groups in the study:control group,lipofectin group,sense oligodexynucleotides(SODN) group,and antisense oligodexynucleotides(ASODN) group. The expression of mdr1 was detected by RT-PCR. Cellular response to irradiation was evaluated by the colony forming test. Methylguanine-DNA methyltransferase(MGMT)protein expression was determined by immunohistochemical staining. Results:ASODN group could significantly decrease the proliferation rate and downregulate the expression of MGMT of irradiated nasopharyngeal carcinoma cells(vs lipofectin group or SODN group,P
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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 Radiosensitivity is highly correlated with DNA doub le stra nd breaks(DSBs). The repair of human DSB is possible mainly through nonhomologou s DNA end joining(NHEJ)pathway. This study was designed to determine the relat ionship between expression of 70Ku/ 80Ku prot ein (a modulating subunit of DNA-PK, which is an important component in NHEJ pathway) and radiosensitivity of nasoph aryngeal carcinoma cell lines. Methods Radiosensitivity parameters of the nasoph aryngeal carcinoma cell lines were obtained by the colony forming assay and radi ation dose-survival curve was done by linear quadratic model. Western blot was p erformed to determine the expression of 70Ku and 80 Ku in total extract of CNE1 a nd CNE2 at baseline level、different time after 4?Gy irradiation、12 h after di ff erent doses of irradiation. To perform a semi-quantitated assay of protein expr e ssion, the optic density (OD) value of each band was estimated by automatic imag e analysis system. Results Surviving fraction of CNE1 was higher than those of CNE2 at each dose point. Mean inactivation dose was higher in the CNE1(2.35)t han that in the CNE2(1.11), but the quantity of 70Ku/ 80Ku in either cell line was similar at both baseline and postirradiation levels. Conclusions CNE2 is mo re radiaosensitive than CNE1; as there was no marked correlation observed betwee n the quantity of Ku protein and radiosensitivity of the nasopharyngeal carcinom a cell lines. X-irradiation cannot result in any change in total quantity of Ku protein.
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Objective To examine the variation of protein expression in nasopharyngeal carcinoma cell lines with different biological characteristics and to identify the radiobiological associated proteins. Methods Biological characteristics of 5-8F and 6-10B were compared by flow cytometry assay after irradia- tion. The total proteins of 5-8F and 6-10B were separated by immobilized pH gradient(IPG) IEF-SDS two- dimensional gel electrophoresis technique. The differentially expressed proteins were cut from the gel and di- gested into peptides for MALDI-TOF MS and the Q-TOF mass spectrometric analysis. Identification of pro- tein was made through searching in protein sequence database. Protein expressions were examined by western blot and immunohistochemistry method. Results Nine most differentially expressed proteins between 5-8F cell and 6-10B cell were identified, p73 and CK19 expression examined by western blot were conformal with that by proteomic method, p73 expression in 5-8F cell was higher than in 6-10B cell. CK19 expression in 6- 10B cell was higher than in 5-8F cell. Conclusion Differentially expression of proteins exist in nasopha- ryngeal carcinoma cell lines with different biological characteristics. These proteins may be associated with cell radiobiological characteristic with the p73 as a potential biomarker.
