Expression Of The Melanoma-associated Antigen-A3 (MAGE-A3) In Cervical Cancer Tissues And Cervical Cancer Cells

Objectives:To further elucidate the expression of melanoma-associated antigen-A3(MAGE-A3)gene in cervical cancer tissues,cervical cancer cell lines Hela,SiHa,C-33 A and normal cervical cell line End1/E6E7 in the expression of cells,explore the possibility of tumor-specific antigen MAGE-A3 as a specific target for cervical cancer immunotherapy,and provide a model for the mechanism of research and lay basis.Methods:The cervical tissue of the first gynecological surgery patients from the Department of Gynecology,Affiliated Hospital of Hebei University was selected from January 2015 to January 2017.Collected archived wax specimens,including 20 normal cervical tissues and cervical cancer specimens in 40 cases.All cases were confirmed by pathological diagnosis,and all patients were excluded from other malignant tumor diseases or acute and chronic diseases.At the same time,The cervical cancer cell lines HeLa,SiHa,C-33A and the normal control cell line End1/E6E7 were cultured.Immunohistochemical SP method was used to detect the expression of MAGE-A3 in 40 samples of cervical cancer tissue and 20 samples of normal cervical tissue.The expression of MAGE-A3 mRNA in three different cervical cancer cell lines and normal cervical cell line was detected by real-time fluorescence quantitative PCR(RT-qPCR).The expression levels of MAGE-A3 protein in three different cervical cancer cell lines-HeLa?SiHa and C-33 A and normal cervical cell line were detected by Western Blot.Result:1.Immunohistochemistry showed that MAGE-A3 was mainly expressed in the cytoplasm and was pale yellow to tan.The positive expression rates of MAGE-A3 in normal cervical tissues and cervical cancer tissues were 0%(0/20)and 70%(28/40),respectively.MAGE-A3 was negatively expressed in 20 cases of normal cervical tissues,28 cases were positive in 40 cases of cervical cancer tissues,and 12 cases were negative.The expression of cervical cancer group was higher than that of normal cervical group,the difference was statistically significant(?~2=26.25,P<0.05).2.Real-time quantitative PCR showed that the mRNA level of MAGE-A3 was detected by GAPDH mRNA as an internal reference standard.The relative quantitative values ofEnd1/E6E7,HeLa,SiHa and C-33 A were 0.124±0.003,0.973±0.004,0.902±0.030 and 0.943±0.017,respectively.The relative expression of MAGE-A3 mRNA was significantly higher in the cervical cancer cell group than in the normal cervical cell group.Statistical analysis showed that there was a statistically significant difference between the cervical cancer cell group and the normal cervical cell group(P<0.05).There was no significant difference between the three cervical cancer cells(P>0.05).3.Western blotting showed that the expression of MAGE-A3 protein in cervical cells showed that MAGE-A3 protein was similar to mRNA in three cervical cancer cells and normal cervical cells.The relative gray values of End1/E6E7,HeLa,SiHa,C-33 A were0.094±0.029,1.005±0.056,0.975±0.026 and 0.984±0.032,respectively.The relative expression of MAGE-A3 protein in cervical cancer cells was significantly higher than that of normal cervical cells.Statistical analysis showed that there was a statistically significant difference between the cervical cancer cell group and the normal cervical cell group(P<0.05),but there was no significant difference between the three cervical cancer cells.Conclusion:MAGE-A3 is highly expressed in cervical cancer tissues and cervical cancer cells,suggesting that MAGE-A3 may play an important role in the development of cervical cancer.It can be seen that MAGE-A3 may be an ideal target for cervical cancer immunotherapy and can also help early diagnosis of cervical diseases.In the future,it is worthwhile to use it to prepare a multi-gene combined modified tumor genetic engineering vaccine,to accurately diagnose and judge the prognosis,and to perform functional targeted therapy in tumor diseases.Therefore,the study of MAGE-A3 gene has important clinical application value for further understanding the biological behavior of cervical cancer cells and prognosis.