| Objective: Hepatocellular carcinoma is the third most common cause of cancer-related death worldwide,its incidence trend is rising in developing countries.Surgery is the most effective treatment of hepatocellular carcinoma,but only 10%-30% of the patients have surgical indications when they were diagnosed with hepatocellular carcinoma.HCC patients often require blood transfusion during liver resection or liver transplantation surgery because of massive blood loss.There are many disadvantages of allogeneic transfusion,such as adverse transfusion reaction,immune suppression,and disease transmission,especially for tumor patients,homologous transfusion may cause a recurrence of tumor.Therefore,autologous transfusion was recommended for liver cancer patients,but it is still a controversial problem,because some scholars worry about that contaminated blood may contain tumor cells can cause distant metastasis of tumor.So,in order to explore the inactivation efficacy of the gamma rays on the residual hepatoma cells in the blood of HCC patients,and to find a suitable dose of irradiation to make autologous transfusion more safe and effective,we studied the effect of gamma rays on the proliferation,migration,invasion and apoptosis of HCC.Methods: 1)Human hepatoma carcinoma cell lines(SMMC7721 and Hep G2)were cultured in vitro.2)HCC were treated with different doses of gamma rays,their proliferating function was assessed by CCK8(cell counting kit-8)assay, the absorbance value can reflect the relative number of living cells;the migration ability of cells was assessed by wound healing assay,the distance of migration can represent the migration ability of cells;the cell invasion ability was assessed by the transwell assay,and the expression of MPP-9 also showed the invasion ability of cells,the more cells invaded and the more MMP-9expressed,the stronger the cell invasion ability;annexin V/PI staining assay and the expression of P53 were used to reflect the apoptosis of cell;3)The data were analyzed by software SPSS 17.0,P<0.05 declared that the difference is statistically significant.Results: 1)The cells lost their normal morphology after they were treated with the gamma rays,cell deformation,floating,incomplete of the cell membrane,vacuoles in cytoplasm,cell fragmentation and a series of changes.2)After being treated with the gamma rays 24 h,48h and 72 h,the ability of cell proliferation was significantly inhibited,the cell survival was inversely proportional to the time and the dose of gamma rays,with the dose and time increasing,cell survival decreased,the difference was statistically significant(P<0.01);3)The gamma rays had an inhibitory effect on migration of cells,the results of wound healing assay(24h)showed that with the dose increased,the distance of cell migration decreased,the difference was statistically significant(P<0.01);4)The gamma rays had an inhibitory effect on the ability of cell invasion,with the increasing of dosage,the number of invaded cells gradually decreased,the difference was statistically significant(P<0.01);Western Blot showed that after irradiation,the expression of protein MPP-9 decreased (P<0.01);5)Gamma rays inhibited the colony formation,start from 20 Gy,there was no colony formation.6)Flow cytometry showed that the early apoptosis rate increased with the rising dose of gamma rays(P <0.01),Western Blot results showed that the expression of P53 increased after the cells were treated with gamma rays(P<0.01).Conclusion: Gamma rays can effectively inhibit the proliferation,migration and invasion of hepatocellular carcinoma cells,accelerate the cell apoptosis,and can effectively inactivate hepatoma carcinoma cells.We recommended that the application of 40 Gy gamma rays in the autologous blood transfusion for HCC patients to improve the safety and effectiveness of auto transfusion. |
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