The Experimental Study On “Deep Sea Water” Combined With Hyperthermia To Treatment The Gastric Cancer

[Purpose]To investigate the effects of PDSW combined with hyperthermia on the related gene expression and protein expression in human gastric cancer cell line SGC-7901 in vitro and in vivo,thus identifying the inhibitory effect of PDSW combined with WBH on human gastric carcinoma cell SGC-7901 growth and its mechanism.[Method]The first part of this study was to explore the anti-cancer effect and its mechanism of PDSW combined with whole body hyperthermia.In in vitro experiments,cultured SGC-7901 cells were randomly divided into control group,PDSW group and normal saline group,with the control group not treated,the PDSW group treated with PDSW and the normal saline group added normal saline of the same quality as PDSW.After being incubated for 24 hours,all the cells were either incubated in 37? conventional condition,exposed the cells to 40? hyperthermia for 6 hours,or exposed the cells to 43? hyperthermia for 1 hours,respectively.After temperature was recovered,growth curves were drawn for gastric cancer cells at day 0,day 1,day 2 and day 3 and MTT assay was performed to measure cell clone formation rate and cell apoptosis rate.What's more,we carried out real-time fluorescent quantitative PCR assay to detect gene expression of PTEN,NF-?b,COX-2,Ki67,Bcl-2,surviving,Vimentin,E-cadherin,Caspase-3,P53,P21 and P16 in cell samples of each group as well as western blot to identify protein expression of PTEN,NF-?b,COX-2,Ki67,Bcl-2,surviving,Vimentin,E-cadherin,Caspase-3,P53,P21 and P1 in cell samples of each group.The second part of this study aimed at exploring the inhibitory effect of PDSW combined with hyperthermia on subcutaneous xenotransplanted tumor from SGC-7901 cells in nude mice.First,the SGC-7901 cells were injected into nude mice,establishing the subcutaneous xenotransplanted mice model of SGC-7901 human gastric cancer.The nude mice were randomly divided into 5 groups.When the tumor grew to a diameter of 1cm,treatments of H20+RT,H20 +40?,H20+43?,PDSW+40?,PDSW+43? treatment were applied respectively.We further drew the growth curve of nude mice tumor,and inspected the tumor tissue sections.RNA was extracted from the tissues to facilitate the real-time fluorescence quantitative PCR assay to detect gene expression of PTEN,NF-?b,COX-2,Ki67,Bcl-2,surviving,Vimentin,E-cadherin,Caspase-3,P53,P21 and P16 in tumor tissue of each group and western blot to identify protein expression of PTEN,NF-?b,COX-2,Ki67,Bcl-2,surviving,Vimentin,E-cadherin,Caspase-3,P53,P21 and P1 in tumor tissue of each group.[Results]In the first part of the experiment,the study on effect of PDSW combined with hyperthermia in human gastric cancer cell SGC-7901 on cell activity and cell apoptosis showed that PDSW had inhibitory effect on the proliferation ability of human gastric cancer cell SGC-7901 in normal body temperature(37?);and the inhibitory effect of PDSW on the proliferation ability of human gastric cancer cell SGC-7901 was more obvious when used in combination with hyperthermia.What's more,the effect of PDSW on cell apoptosis of SGC-7901 cells presented a similar relationship.PDSW combined with the hyperthermia will increasing the gene and protein expression of PTEN,Caspase-3,tumor suppressor gene p53,p21 and p16 in SGC-7901 cells,and decreased gene and protein expression of Ki67,COX-2,Bcl-2,NF-?b,survivin and vimentin.Therefore,it could effectively inhibit the proliferation of gastric cancer cell.However,the abnormal gene and protein expression of E-cadherin,which,like vimentin,is a key regulator in EMT phenomenon,needs to be further verified in vivo.Therefore,it is speculated that PDSW combined with hyperthermia can effectively inhibit the proliferation and promote apoptosis of human gastric cancer cell line SGC-7901.In the second part of the study,the result regarding the inhibitory effect of PDSW combined with hyperthermia on tumor in vivo showed that under normal body temperature(37?),PDSW had inhibitory effect on the proliferation of subcutaneous xenotransplanted tumor in mice.Again,when with the copresence of hyperthermia(40? for 6 h or 43? for 1 h),the inhibitory effect of PDSW on the proliferation ability of xenotransplanted tumor in mice was more significant.Similar to the outcome after the treatment of PDSW combined with hyperthermia on gastric cancer cell SGC-7901,the same treatment on mice with transplanted human gastric cancer would lead to significant inhibition of protein expression of NF-?b,COX-2,Ki67,B,Bcl-2,survivin and Vimentin,and also significant up-regulation of protein expression of PTEN,E-cadherin,Caspase-3,P53,P21 and P16.Through regulation proapoptotic gene and apoptosis-suppressing gene expression,PDSW combined with hyperthermia could effectively inhibit the proliferation of gastric carcinoma cells.Therefore,PDSW combined with hyperthermia could approach the goal of treating gastric cancer by effectively inhibit the proliferation of tumor cells in mice transplanted tumor and promote their apoptosis.[Conclusion]Either in vivo or in vitro,under normal body temperature(37?),PDSW has inhibitory effect on the proliferation ability of human gastric cancer cell SGC-7901.This inhibitory effect is more obvious when PDSW treatment is combined with hyperthermia(40? for 6 h or 43 ? or 1 h).PDSW combined with hyperthermia can activate up-regulation of the protein expression of PTEN,E-cadherin,Caspase-3,P53,P21 and P16,and inhibit gene and protein expression of NF-?b,COX-2,Ki67,Bcl-2,survivin and Vimentin in both SGC-7901 gastric cancer cell and transplanted tumor tissue.Therefore,PDSW combined with hyperthermia could approach the goal of treating gastric cancer by effectively inhibit the proliferation of human gastric cancer cell line SGC-7901 and promote their apoptosis.