Research On The Tumor Suppressor Function Of Testin Gene In Non-small Cell Lung Cancer

Objective:The objective of this research is to explore the effect of Testin gene overexpression on nude mice transplantation tumor of lung cancer A549 cells.Methods:1.The Testin gene overexpression vector and empty vector were constructed and used to transfect A549 cells using lentivirus transfection technique to establish Testin overexpression cells(A549-Testin-OVER)and control cells(A549-control).Stable expression cells were screened out through puromycin;2.The transfection efficiency was measured by semi-quantitative PCR and Western Blot analyses in A549-Testin-OVER cells;3.In order to investigate the tumor suppressor function of Testin gene in vivo,A549-Testin-OVER cells,A549-control cells and untreated A549 cells were subcutaneously inoculated into the fossa axillaris of nude mice.Paid attention to the status of nude mice everyday and measured the tumor length(L)and width(W)twice a week for 5 weeks.Draw the tumor growth curve according to the volume of the tumors.Kill the mice 5 weeks later,the tumor tissues were collected and weighed to evaluate the function of inhibiting the tumorigenicity in vivo;4.The protein levels of Testin in the xenografts were examined by immunohistochemistry.Results:1.The Testin gene overexpression vector and empty vector were successfully transfected into A549 cells.Stable expression cells were screened out through puromycin.Good transfection efficiency of Testin gene overexpression was confirmed by semi-quantitative PCR and Western Blot;2.The nude mice injected with A549-Testin-OVER cells formed tumors later than control groups.Furthermore,the tumor growth was also greatly reduced and smaller tumors were formed compared with other groups within 5 weeks;3.The positive rate of Testin protein(98.32±1.76)% in the tumors of A549-Testin-OVER group was siginificantly higher than A549-control group(22.92±21.46)% and untreated A549 group(20.14±22.54)%(P <0.05).Whereas,there was no significant difference between the two control groups(P >0.05).Conclusions:Testin gene,as a tumor suppressor,plays a key role in the inhibition of non-small cell lung cancer cell xenograft formation and growth in vivo.