PURPOSETo investigate the radiosensitization by prodrug and CD-TK double suicide gene therapy system in adenoid cystic carcinoma cells (ACC-2).METHODSThe eukaryotic expression plasmids pIRES-CD and pIRES-TK were introduced into ACC-2 cells by electroporation. Then ACC-2 cells stably expressing CD and TK gene were obtained by 10-day positive selection with 400 micro g/mL G418 . The total RNA was extracted and the expression of the CD and TK gene in transfected ACC-2 cells was identified by RT-PCR. The positive transfected ACC-2 cells were treated with radiotherapy of different dose (0,2,4,6,8,10 Gy) and prodrug system in aerobic and anoxic condition. Then cell clone formation assay was used to study the radiosensitization by CD-TK double suicide gene therapy and prodrug system in ACC-2.The data was analyzed by multiple factor ANOVA using SPSS11.5 software package.RESULTSRT-PCR analysis demonstrated that CD and TK genes were effectively expressed in ACC-2 cells. With the increased of X-ray dose, the colony forming rate dropped significantly after radiotherapy. In aerobic condition, the survival fraction of group ACC-2/CD-TK+prodrug were significantly lower than that of group ACC-2 and group ACC-2/CD-TK with the same dose (P<0.05). In anoxic condition, the survival fraction of group ACC-2/CD-TK+pro-drug was significantly lower than that of experimental group ACC-2 and group ACC-2/CD-TK with the same dose (P<0.05). The colony forming rate in aerobic condition was significantly lower than that in anoxic condition of the same cell group and dose.CONCLUSIONThe radiosensitivity and the killing effect of X ray to ACC-2 cells can be increased by CD-TK double suicide gene therapy and the prodrug system.

2007-12-01·BMC Cancer2区 · 医学

In vitro angiogenesis and expression of nuclear factor κB and VEGF in high and low metastasis cell lines of salivary gland Adenoid Cystic Carcinoma

2区 · 医学

ArticleOA

作者: Bin Peng ; Jiali Zhang

BACKGROUNDAdenoid cystic carcinoma is a high malignant carcinoma characterized by intensive local invasion and high incidence of distant metastasis. Although many reports have demonstrated that angiogenesis has played an important role in tumor metastasis, the relationship between metastasis characters and angiogenesis ability in high and low metastasis cell lines of Adenoid cystic carcinoma has rarely been reported. The present study aimed to compare the angiogenesis ability of ACC-M (high metastasis) and ACC-2 (low metastasis) cell lines in vitro. Furthermore, the activity of nuclear factor kappaappa B and the expression of vascular endothelial growth factor (VEGF) in ACC-2 and ACC-M were also detected.METHODSElectrophoretic mobility shift assay was used to detect nuclear factor kappaappa B activity. Semi-quantitative RT-PCR was used to quantify the mRNA level of VEGF. Immuofluorescence double staining and semi-quantitative confocal laser scanning analysis was carried out to detect nuclear factor kappaappa B nuclear localization and staining intensity of VEGF. The angiogenesis ability of ACC-M and ACC-2 was compared by an in vitro three-dimensional angiogenic model assay. The vector transfection assay was performed to transfect the PCMV-IkappaBalphaM vector into ACCs cell lines expressing the phosphorylation defective IkappaBalphaM.RESULTSNuclear factor kappaappa B activity and the rate of nuclear factor kappaappa B nuclear localization in ACC-M was significantly higher than that in ACC-2. Moreover, ACC-M exhibited higher mRNA and protein levels of vascular endothelial growth factor than ACC-2. VEGF mRNA expression was effectively decreased by inhibition of nuclear factor kappaappa B activity. Furthermore, ACC-M could remarkably stimulate the migration and tube formation of endothelial cells and induce The umbilical vein endothelial cells sprouting into the gel matrix.CONCLUSIONThese results implicated that ACCs cells with higher metastasis feature might present greater angiogenesis ability.

2005-12-01·International Journal of Oral and Maxillofacial Surgery3区 · 医学

Integrins in metastatic adenoid cystic carcinoma

3区 · 医学

Article

作者: J. Jones X.-T. Xie ; A.J. Lyons

Adenoid cystic carcinoma exhibits a diverse clinical behaviour from very localised invasion to widespread metastatases. Recently two adenoid cystic carcinoma cells lines have been isolated and cultured which distinctly show either a low metastatic tendency (Acc-2) or a highly metastatic behaviour (Acc-M). It was hypothesised that these two types of behaviour may at least in part be explained by the different integrin profile on the cells' surface membrane. The integrins represent the largest known family of cell adhesion molecules and a number of tumour cell processes have been shown to be dependant on their integrin expression. Adenoid cystic carcinoma cells of both types were obtained and successfully cultured. These were then subjected to integrin analysis by a number of monoclonal antibodies to alpha(1), alpha(2), alpha(3), alpha(v), alpha(6), beta(1), alpha(v)beta6, alpha(v)beta4 using a fluorescence activated cell sorter. Although there was an apparent difference of integrin expression in the Acc-M group as compared to the Acc-2 group, this was not statistically significant. It is still possible though, that this might account for the differences in behaviour of the two cell lines, and the relationship between integrins, and invasion and metastases in other tumours is discussed. The pattern of integrin expression in Acc may be prognostically significant and useful for treatment planning in the future.

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