Effect Of PTTG1 In Invasion And Metastasis Of Endometrial Carcinoma

Objective:to explore the effect of PTTG1 shRNA interference plasmid on proliferation,invasion and metastasis of endometrial carcinoma.Methods:1.Cell culture:including Cell Recovery,Cleaning,Passage,Cryopreservation,Cell count and so on.2.Structure three PTTG1 sh RNA plasmid(PTTG1-homo-509;PTTG1-homo-922;PTTG1-homo-596).3.Transfection the PTTG1 sh RNA plasmid to human endometrial carcinoma Ishikawa(ISK)cells under the effect of liposome.4.To observe the green fluorescent protein expression by fluorescence microscope,determine the cell transfection efficiency.5.To detect the PTTG1 changes of protein level by Western blot method.6.To test the effect of PTTG1 on growth of Ishikawa cells by MTT Cell proliferation experiment.7.To detect the effect of PTTG1 on migration and invasion of human endometrial carcinoma Ishikawa cells by Transwell experiment.Results:1.After transient transfection ISK cells 36 hours,we observed that the green fluorescent was about 75% in ISK cells by fluorescence microscope,that indicated that cell transfection experiment was successful.2.Compared to the negative control group and the normal group,PTTG1 expression in the three interferential group were obviously decreased as show by Western Blotting(P < 0.05).This means that in the three interference group had significantly inhibited the PTTG1 protein expression,namely the three targeting shRNAinterference plasmid of PTTG1 gene were effective3.Cell proliferation abilities of the interferential group were both significantly lower than the control group and the normal group by MTT cell proliferative assay(P<0.05).4.Cell migration and invasion abilities of the interferential group were both significantly lower than the control group and the normal group by Transwell experiment(P<0.05).Conclusions:1.PTTG1 shRNA interference plasmid can significantly inhibit PTTG1 expression in ISK cells and abilities of cell proliferation,invasion and metastasis.2.PTTG1 gene played an important role in the occurrence and development of endometrial cancer.