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Objective:To investigate the effect of BMAL1 gene on the proliferation, migration and invasion ability of radiation-resistant nasopharyngeal carcinoma cell line (5-8FR) and the molecular mechanism. Methods:A multi-target click model was constructed for radiation-resistant nasopharyngeal carcinoma cell line 5-8FR by low-dose fractionated irradiation, and the results of clone formation assay were used to fit the multi-target click model and calculate the sensitization ratio of radiotherapy. The expression levels of PI3K/Akt/MMP-2/9 signaling pathway-related proteins in 5-8FR and control 5-8F cell lines were detected by Western blot. The overexpression and knockdown vectors of BMAL1 gene were constructed and transfected with 5-8F and 5-8F cell lines, respectively. The BMAL1 gene overexpression (pcDNA-BMAL1) and its control (pcDNA) and interference (BMAL1-shRNA) and control (con-shRNA) cell lines were stably transfected with nasopharyngeal carcinoma cell line 5-8F and radiation-resistant cell line 5-8FR, respectively. Western blot was performed to verify the infection efficiency and detect the changes of PI3K/Akt/MMP-2/9 signaling pathway-related proteins after overexpression or interference of BMAL1 gene in both groups of cells. CCK-8 assay, cell scratch test and Transwell chamber test were conducted to investigate the proliferation, migration and invasion capabilities of 5-8FR cell line after overexpression or interference of BMAL1 gene. Results:BMAL1 gene expression was down-regulated, and those of PI3K/Akt pathway proteins and downstream related molecules of MMP-2 and MMP-9 were up-regulated, and TIMP-2 and TIMP-1 expression was down-regulated in nasopharyngeal carcinoma radiation-resistant cell lines. Overexpression of BMAL1 gene inhibited the expression of PI3K/Akt pathway proteins and downstream related molecules of MMP-2 and MMP-9, promoted the expression of TIMP-2 and TIMP-1, and inhibited the proliferation, migration and invasion capabilities of radiation-resistant nasopharyngeal carcinoma cells, while interference with BMAL1 gene yielded the opposite results. Conclusions:BMAL1 gene can reverse the expression of PI3K/Akt/MMP-2/9 signaling pathway-related proteins in radiation-resistant nasopharyngeal carcinoma cell lines and inhibit the proliferation, migration and invasion capabilities of radiation-resistant nasopharyngeal carcinoma cell lines.
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Objective To study the killing effect of cytokine-induced killer cells (CIK cells) on human lung adenocarcinoma cell line (A549) and the lung adenocarcinoma' s radiation resistant cell line (A549RR).Methods Peripheral blood mononuclear cells (PBMC) of healthy volunteers were stimulated by different cytokines,and were induced into killer activity CIK cells.The phenotype of CIK cells were analyzed by flow cytometer.A549 and A549RR cell lines were cultured separately with the CIK cells.The absorbance value (A) of the cells was measured by CCK8,and the killing rates of all cells which were cultured for 24 and 48 hours with the CIK were calculated.Results The rate of CD3+ CD56+ cell was 45.8 ％ after culture for 14 d.The killing rates of CIK cells to lung adenocarcinoma A549 cells and its radiation resistant cells A549RR were increased with the rise of the ratio of effective cells to target (5∶1-40∶1) and the increasing of culturing time (all P ＜ 0.001).The killing effect of CIK to A549 and A549RR cells had no obvious difference in the same culturing time and the same ratio of effective cells to target(all P ＞ 0.05).Conclusion CIK cells have strong anti-tumor effect against lung adenocarcinoma and its radiation resistant cells with high clinical application value.
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It is a mature technology producing single-use syringes by polypropylene(PP). But the traditional sterilization process for single-use syringe pollutes the environment, and the residues endanger the health of users. Irradiation sterilization, instead of traditional sterilization process, is the developing trend. However, the traditional PP does not have the ability of radiation resistance. These methods can improve the radiation resistance ability of PP effectively by adding the stabilizer or floating agent. Toughening or temperature treatment can have the same function. Radiation-resistant PP is introduced in this paper. Furthermore, the developing trends for the materials are discussed.
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How follicular T-helper (Tfh) cells develop is incompletely understood. We find that, upon antigen exposure in vivo, both naïve and antigen-experienced T cells sequentially upregulate CXCR5 and Bcl6 within the first 24 h, relocate to the T-B border, and give rise to phenotypic Bcl6(+)CXCR5(+) Tfh cells before the first cell division. CXCR5 upregulation is more dependent on ICOS costimulation than that of Bcl6, and early Bcl6 induction requires T-cell expression of CXCR5 and, presumably, relocation toward the follicle. This early and rapid upregulation of CXCR5 and Bcl6 depends on IL-6 produced by radiation-resistant cells. These results suggest that a Bcl6(hi)CXCR5(hi) phenotype does not automatically define a Tfh lineage but might reflect a state of antigen exposure and non-commitment to terminal effector fates and that niches in the T-B border and/or the follicle are important for optimal Bcl6 induction and maintenance.
Subject(s)
Animals , Mice , CD40 Ligand , Metabolism , Cell Differentiation , Physiology , DNA-Binding Proteins , Metabolism , Inducible T-Cell Co-Stimulator Protein , Metabolism , Interleukin-6 , Metabolism , Proto-Oncogene Proteins c-bcl-6 , Receptors, CXCR5 , Metabolism , T-Lymphocytes, Helper-Inducer , MetabolismABSTRACT
[ ABSTRACT] AIM:To compare the differences of the genome-wide methylation levels and methylated regions be-tween nasopharyngeal carcinoma ( NPC) cells in the same genetic background but different radiation resistance ( CNE-2 cells and CNE-2R cells).METHODS:Using the method which was developed by Doctor Zhao Cun-you, based on using methyl-sensitive restriction enzyme to measure the genome-wide methylation levels.In addition, MeDIP-Seq was used to analyze the methylated regions in 6 gene functional elements, including the upstream 2k sequence, 5’ UTR, coding se-quence, intron, 3’UTR and downstream 2k sequence, between CNE-2 cells and CNE-2R cells.RESULTS:The genome-wide methylation level was approximately 30%lower in CNE-2R cells than that in CNE-2 cells.No obvious difference on the amount of genes and the coverage of the peak in the 6 gene functional elements was observed.However, the methylation pattern of plentiful genes had altered in the gene function elements.CONCLUSION:The genome-wide methylation levels and methylated regions between NPC cells in the same genetic background but different radiation resistance were quite dif-ferent, indicating that the DNA methylation may be associated with NPC radioresistance.
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Xeroderma pigmentosum is rare autosomal recessive disease, characterized by extreme sensitivity to ultraviolet light and leading to development of multiple malignant skin tumors. Among multiple malignant skin tumors, basal cell carcinoma and squamous cell carcinoma are common. Treatments of xeroderma pigmentosum are palliative or radiation treatment, but surgical resection is necessary if xeroderma pigmentosum is transited to malignant skin tumors. We report a case of a 32-years-old female xeroderma pigmentosum patient, who developed radiation resistant basal cell carcinoma on right alar nasi. Radiaton theraphy(300cGy, fractions: 10, total dose: 3000cGy) was not effective, so surgical excision and local skin flap coverage was made and no specific wound complications occurred. During 11 months follow-up period, no evidence of recurrence was found.