Preliminary Study On Anti-Tumor Auxiliary Effect And Mechanism Of Natural Deterium Depleted Water In Xinjiang Real Estate

Objective: Two natural deterium depleted water aquatic products,AKT and GLX,were selected from Xinjiang Real estate to screen their inhibitory effects on the growth of tumor cells(lung cancer cells,liver cancer cells,gastric cancer cells,cervical cancer cells,melanoma cells,pheochromocytoma).Preliminary study on the inhibitory effect of effective products on tumor cells and its mechanism.Methods: 1)In the preliminary anti-tumor screening study of natural deterium depleted water in Xinjiang,CCK-8 and flow cytometry were used to detect deterium depleted water for lung cancer cells(A549),liver cancer cells(HepG2),and gastric cancer cells(AGS).Proliferative activity and apoptosis of cervical cancer cells(Hela),melanoma cells(A375),and pheochromocytoma cells(PC-12);2)The effect of AKT on the apoptosis of human lung cancer cells A549 in Xinjiang real estate was studied by Transwell method.The effect of AKT on the migration ability of A549 cells was detected by RT-PCR and Western-blot method.The effect of water AKT on the expression levels of P53,Bcl-2,Bax,Caspase-3 and VEGF in A549 cells;3)In the study of the anti-tumor effect and mechanism of deterium depleted water AKT on tumor model mice in Xinjiang,A Lewis lung cancer mouse model was established to investigate the(1)tumor inhibition rate,(2)organ index,(3)serum biochemical indicators,etc.Furthermore,the effectiveness of AKT anti-tumor effect was further verified in vivo.(4)Western-blot assay was used to detect the effects of AKT on the expression of P53,Bcl-2,Bax,Caspase-3 and VEGF in lung tissue of Lewis lung cancer model mice.Results: 1)In cell proliferation,GLX groups had no obvious proliferation inhibition effects on melanoma cells,gastric cancer cells,pheochromocytoma,lung cancer cells and cervical cancer cells,and the same proliferation occurred in the control group;The AKT group had no obvious effect on hepatoma cells and cervical cancer cells.The inhibition rate of lung cancer cells,gastric cancer cells,melanoma cells and pheochromocytoma cells in the AKT group was up to 19.42%,11.21%,16.19%and 14.75%.Flow cytometry revealed that AKT mainly affects the early stage of apoptosis of lung cancer cells,melanoma cells and pheochromocytoma cells,and the early stage of gastric cancer cells;2)Transwell results showed that AKT could significantly reduce the invasive ability of lung cancer cells;RT-PCR and Western-blot found that AKT significantly up-regulated the expression of Caspase-3,Bax and P53 genes,and significantly down-regulated the expression of Bcl-2gene.At the same time,AKT can significantly down-regulate the expression of VEGF gene;3)(1)The positive control group had a tumor inhibition rate of 45.32%,a volume fraction of 0.005%.The DDW group had a tumor inhibition rate of 36.66%,a volume fraction of 0.0075%DDW tumor inhibition rate of 16.74%,and a volume fraction of 0.010%DDW tumor inhibition rate of 9.44%.(2)The spleen coefficient and thymus coefficient of the mice in each group were significantly decreased,and the difference was statistically significant(P<0.01).(3)Compared with the model group,the four specific biochemical indicators of serum tumors in each drug-administered group were different;compared with the positive control group,the four specific biochemical indicators of serum tumors were also different.(4)Western-blot results indicated that AKT significantly up-regulated the expression of Caspase-3,Bax and P53 genes,and significantly down-regulated the expression of Bcl-2gene,while AKT significantly down-regulated the expression of VEGF gene.Conclusion: Xinjiang low-altitude aquatic product AKT can inhibit the proliferation of lung cancer cells,gastric cancer cells,melanoma cells and pheochromocytoma cells in vitro,which can improve the survival status of Lewis model mice and improve the immune function of mice.It is suggested that AKT has obvious anti-tumor effect,and its mechanism may be related to up-regulation of P53,Caspase-3,Bax and down-regulation of Bcl-2 protein expression,and at the same time,it can reduce the expression of VEGF.