Establish A Human Lung Adenocarcinoma Resistant Cell Line H1299R And Investigate The Variation Of Radiosensitivity Effected By Interactions Between HIF-1? And MiR-183

Objective: In the anoxic microenviroment, the higher expression of HIF-1? maintain the stability of oxgen and metastation to promote own growth and mestastatis, thus producing radiation resistance leads to treatment failure. Previous studis have found that mi R-183 can control HIF-1?, however the relationship between them and the radiation sensitivity has not been clarified. In this study, On the basis of establishing the model of human lung adenocarcinoma cell line, we observed the changes of cell radiosensitivity and explored the relationship between mi R-183 and HIF-1? in the treatment of radiation resistance by intervening the expression of mi RNA-183.Methods: Human lung adenocarcinoma H1299 radiation resistant cells were induced by conventional dose fractionation and sublethal dose mode. Colony formation assay was used for analysis of radiation resistance. Cell apoptosis and cell cycle were detected using flow cytometry. H1299 radiation resistant cells transfected with lentivirus, then mi R-183, HIF-1? m RNA expression were detected RT-PCR. The CCK-8 method was used to measure cell proliferation.Results: The SF2?D0 and other radiation sensitive parameters of H1299R-2 Gy cells were more than that of the parent cells and H1299R-6 Gy cells. Moreover, the cell survival shoulder area of H1299R-2 Gy cells is the most wide, which has the strongest radiation resistance, so we named H1299 R cells. CCK-8 experiment and flow cytometry results showed that, compared with the parental H1299 cells, H1299 R cells had significant radiation resistance(P<0.05); RT-PCR results show that the levels of mi R-183 and HIF-1? m RNA in H1299 R cells were higher than that of H1299 cells(P<0.05); After transfection of H1299 R cells, the level of HIF-1? m RNA was also relatively low(P<0.05); The growth curve showed that there was no significant difference between the H1299 R cells and the control group in the sensitivity to X rays when the expression of mi R-183 was downregulated(P>0.05).Conclusion: Human lung adenocarcinoma resistant cell lines can be successfully constructed by conventional fractionated irradiation method. Compared with the parent cells, resistant cell lines had stronger radiation resistance for X-ray, which associated with the high expression of HIF-1?. We speculated mi R-183 may affect the expression of tumor cells by regulating the expression of HIF-1?.