The Effect Of Exogenous Tumor Antigen MAGE-A9 And MAGE-A11 On The Growth Of Esophageal Cancer Xenograft In Nude Mouse

The incidence of esophageal cancer is the fourth in the world,according to the statistics of the international agency for research on cancer,in China,as esophageal cancer high-risk area.Currently,immunotherapy has become an anti-cancer new method,which will be the new trend following other major cancer therapies such as surgery,radiation and chemotherapy etal.Mechanism of the immunotherapy is that biological defense mechanism has been activated to against cancer cells.So,the key of immunotherapy is screening for tumor specific antigen.The expression pattens of cancer/testis antigens(CTA)is peculiar that CTA expressed in various cancer cells,but don’t in normal human tissues except the germ cell of testis and placenta.Nevertheless,placenta and testis which don’t expressing human leukocyte antigen(HLA)are immunological privileged organ.So that,CTA are suitable as target antigen for specific tumor immunotherapy.Melanoma-associated antigens(MAGE)families that belong to the CTA expressed in various cancer cells,but don’t in normal human tissues except the germ cell of testis and placenta.MAGE families are classified as two group including MAGE-Ⅰand MAGE-Ⅱ.MAGE-Ⅰare specific tumor antigen and classified as three gene subtype such as MAGE-A、MAGE-B and MAGE-C,among which MAGE-A compose of 12 members,while MAGE-Ⅱ don’t belong to the CTA family because it expressed in normal tissues.In our research,ECA109 cells by gene transfection were inoculated under the skin in nude mice to set up esophageal tumor-burdened model.Then,the growth of xenograft tumors were measured to draw the tumor growth curve on alternate days,and the weight of xenograft tumors were weighed outin the end of the experiment.The excitated fluorescence energy of GFP fluorescence protein was detected under small animals vivo fluorescence imaging system.In order to explore the role of MAGE-A9 and MAGE-A11 which is targets for esophageal cancer immunotherapy in xenograft tumor in vivo.Objective: To explore the role of exogenous MAGE-A9 and MAGEA11 gen in esophageal cancer xenograft in vivo in nude mice.Methods: Eca109 over-expressioned MAGE-A9 and MAGE-A11 gen were inoculated into nude mice subcutaneously,respectively,to set up esophageal tumor-burdened model.Then,the growth situation of xenograft tumors were measured,and the excitated fluorescence energy of xenograft tumor in vivo was detected under small animals vivo fluorescence imaging system,to reflect the activity of nude mice subcutaneous transplantation tumor.Eca109 transfected blank plasmid has been inoculated into nude mice subcutaneously transplantation tumor model as a control.In order to explore the role of MAGE-A9 and MAGE-A11 gens in human esophageal cancer cellsResult:1 The success rate of transfection of control group,p CMV6-AC-MAGEA9-GFP group and p CMV6-AC-MAGE-A11-GFP group was 91.2%,92.06% and 89.83% respectively.2 The xenograft tumor in nude mice growth assumes the circular or elliptic,smooth surface as a whole,and there are two or three disperse tumors in individual animals,while 1 case without tumor in control group.3 xenograft tumor increased gradually since 15 days after cells were inoculated in control group,while increased obviously since 10 days in other two experiment group.It is showed that xenograft tumor over-expressioned MAGE-A9 or-A11 gen could growth promptly in compare with control group on the graph of xenograft tumors growth curve.4 The weight of xenograft tumor in control group,MAGE-A9 group and MAGE-A11 group is 0.29±0.046 g,0.99±0.132 g and 0.69±0.072 g,respectively.It is showed that both MAGE-A9 and MAGE-A11 gen could promotexenograft tumors growth.5 The excitated fluorescence energy of xenograft tumor in vivo in control group,MAGE-A9 group and MAGE-A11 group is 37.76±2.01、 69.93±6.31 and 67.76±7.79(CPS×10~5)respectively.The photo of fluorescence imaging in vivo showed that the fluorescence intensity of transplantion tumors over-expressioned MAGE-A9 or-A11 gen are greater in compare with control group.Conclusion: Exogenous MAGE-A9 and MAGE-A11 gen could promote growth and activity of xenograft tumors in nude mice.