Effects of X-ray on LOX family in gastric cancer cells and human umbilical vein endothelial cells
Objective This study is aimed to compare the effects of different doses of X-ray on expressions of lysyl oxidase(LOX)family members in human poorly differentiated mucoid gastric adenocarcinoma cell(MGC-803)and human umbilical vein endothe-lial cell(HUVEC).Methods MGC-803 and HUVEC were ex-posed to different doses(0 Gy,2 Gy,4 Gy,6 Gy,8 Gy and 10 Gy)of X-ray.The growth of cells was evaluated using cell proliferation as-say,then the Lethal Dose of 50%(LD50)was calculated to determine the dosage for subsequent research.The abundance of mRNA,rela-tive quantity of protein and levels of secretion of LOX family members in two cell lines were detected through real-time PCR,Western blot(WB)and enzyme-linked immunosorbent(ELISA).Enzyme ac-tivity of LOX was tested with Amplex Red hydrogen peroxide assay.The changes of p-p38MAPK and p-ERK1/2 MAPK were analyzed by WB after X-ray exposure.Results The LD50 of X-ray radiation were 8.2 Gy to MGC-803 and 6 Gy to HUVEC.Compared with 0 Gy,the mRNA abundances of LOXL1,LOXL2 and LOXL4 of gastric cancer cells MGC-803 were down-regulated,but the LOX mRNA was up-regulated after radiation by 2 Gy,4 Gy,6 Gy X-ray(P＜0.05).The relative quantity of proteins and levels of enzyme secretion of LOX and LOXL4 were down-regulated after radiation by 2 Gy X-ray(P＜0.05).The relative quantity of proteins and levels of en-zyme secretion of LOX,LOXL1,LOXL2 and LOXL4 were down-regulated after radiation by 4 Gy X-ray(P＜0.05).the relative quanti-ty of proteins and levels of enzyme secretion of LOXL1 were down-regulated after radiation by 6 Gy X-ray(P＜0.05).The abundance of mRNA,relative quantity of protein and levels of secretion of LOXL3 were up-regulated under 2 Gy radiation(P＜0.05).Compared with 0 Gy,the levels of mRNA,protein and secreting enzyme of LOXL1 and LOXL3 of HUVEC were significantly up-regulated after ir-radiation by 2 Gy X-ray(P＜0.05).The levels of mRNA,protein and secreting enzyme of LOX,LOXL1,LOXL3 and LOXL4 were up-regulated after irradiation by 4 Gy X-ray(P＜0.05).The levels of mRNA,protein and secreting enzyme of LOXL2 and LOXL4 were up-regulated after irradiation by 6 Gy X-ray(P＜0.05).LOX enzyme activity of HUVEC was enhanced after 2 Gy and 6 Gy X-ray irra-diation(P＜0.05).The expression of p-p38,p-p42 and p-p44 in MGC-803 cells were upregulated after 2 Gy and 6 Gy irradiation(P＜0.05),and the expression of p-P38,p-P42 and p-P44 in HUVEC were up-regulated after irradiation of 2 Gy,4 Gy and 6 Gy irradia-tion(P＜0.05).Conclusion Two cell lines exhibit differently biological behaviors after X-ray exposure.The production,secretion and enzyme activity of LOX and its family members were up-regulated in HUVEC after X-ray exposure.But the production and secre-tion were down-regulated in MGC-803 after X-ray exposure,except LOXL3.
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