| AIM:Experimented on clinical specimens in the early stage of the study of CSF and the method of immunohistochemical analysis CSF-1R in nasopharyngeal carcinoma and nasopharyngitis differentially expressed on the basis of further by building CSF-1R cell line expression and interference,as well as the experiment from the cell level to explore nasopharyngeal carcinoma cells in vitro radiation resistance ability of cell proliferation,apoptosis,invasion and metastasis.We hope for the treatment and prognosis of nasopharyngeal carcinoma found a new predictor,hope this can also be new targets for the treatment of patients with nasopharyngeal carcinoma.Methods:1.The 6-MV X-ray linear accelerator according to 0Gy,2Gy,4Gy,6Gy,8Gy dose was divided into 5 groups,respectively irradiation CSF-1R(macrophage colony stimulating factor 1 receptor)positive cell line 5-8F and CSF-1 negative cell line 6-10 B,continue to develop each group respectively cells after irradiation,the establishment and each dose groups,each cell lines corresponding to the radiation resistance of the cell lines.2.CCK-8 method to detect the CSF-1R positive cell line 5-8F and negative CSF-1 cell line 6-10 B proliferation capacity,and testing according to the radiation dose group of 5-8 f cell lines of radiation resistance cell line and 6-10 B cell lines of radiation resistant strains between groups of proliferation.3.Detection of CSF of clone formation experiment 1 positive cell line 5-8F and r negative CSF – 1R cell line 6-10 B proliferation capacity,and testing according to the radiation dose group of 5-8F cell lines of radiation resistance cell line and 6-10 B cell lines of radiation resistant strains between groups of proliferation?4.Using flow cytometry to detect positive CSF – 1R cell line 5-8F and negative CSF – 1R cell line 6-10 B of apoptosis,as well as the test according to the radiation dose group of 5-8F cell lines of radiation resistance cell line and 6-10 B cell lines of radiation resistant strains between groups of apoptosis.5.CSF-1R in Transwell experiment testing positive cell line 5-8F and negative CSF-1R cell line 6-10 B ability of invasion and metastasis,as well as the test according to the radiation dose group of 5-8 f cell lines of radiation resistance cell line and 6-10 b cell lines of radiation resistant strains between groups of ability of invasion and metastasis?6.Using RT-PCR detection of CSF-1R positive cell line 5-8F and CSF-1R negative cell lines and 6-10 B test according to the radiation dose group of 5-8F cell lines of radiation resistance cell line and 6-10 B cell lines of radiation resistant strains between groups of P53,...The expression level.7.Through a technique called RNA interference to build 6-10 B cell lines CSF-1R siRNA slow virus expression vector.Result:1.was detected in vitro 5-8F cell lines(CSF-1R+)and 6-10 B cell lines(CSF-1R-)cell proliferation,apoptosis,migration,invasion and metastasis ability difference: proliferation,experiments show that cell line 5-8F value is higher than cell lines,6-10 B apoptosis shows that there are differences between the two kinds of cell apoptosis is,the difference was statistically significant.2.through the in vitro experiment testing after exposure to X-ray 5-8F cell lines(CSF-1R+)and 6-10 B cell lines(CSF-1R-),collect different doses and different time points of cells proliferation,apoptosis,migration,invasion and metastasis ability of change,and detection and proliferation,apoptosis,migration,invasion and metastasis related factors:2.1 the CCK-8 and clone forming experiments results show that the X-ray have obvious inhibitory effect on cell proliferation,cell in the short term for X line resistance is not obvious,the forward for X ray resistance cell line 5-8 f above 6-10 B.2.2 proliferation related factor P53 in cell 2Gy x-ray after radiation exposure,expression is not obvious;Apoptosis related factor FasL in cell line 5-8F cells to accept 2Gy x after radiation exposure,there are differences in expression,but in cell lines 6-10 B did not change significantly.3.by RT-PCR from the level of gene detection 5-8 f cell lines(CSF-1R+)and 6-10 b cell lines(CSF-1R-)cell line proliferation,apoptosis,migration,and the expression of invasion and metastasis related factors:3.1 proliferation related factor P53 in 5-8F cell lines(CSF-1R+)and 6-10 B cell lines(CSF-1R-)the expression did not see obvious difference;3.2 apoptosis related factor FasL in 5-8 f cell lines(CSF-1R+)and 6-10 b cell lines(CSF-1R-)the expression did not see obvious difference.4.successful build CSF-1R too slow virus expression vector.For the next step research CSF-1R negative cell line transfection CSF-1R cell function changes after slow virus expression vector.Conclusion:1.different nasopharyngeal carcinoma cell line proliferation and apoptosis.2.Different nasopharyngeal carcinoma cell line proliferation and apoptosis before and after the exposure to X-ray ability difference,and CSF-1R+ cell lines and CSF-1R-cell line proliferation and apoptosis before and after irradiation ability change level is different.3.The CSF-1R expression and radiation resistance of the nasopharyngeal carcinoma cell lines have a certain correlation,its mechanism is complex,the specific reason remains to be further research4.Successful build CSF-1R too slow virus expression vector,for the next step research negative CSF-1R cell line transfection CSF-1R slow virus expression vector changes in cell function. |
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