Impaction Of Fragments Of EBV, HPV And PTEN Tumor Suppressor On Genesis Of Lung Cancer

We have detected DNA sequence of Epstein-Barr virus in 32 cases out of 80 (40%) lung tumor samples of various histological type and Human papilloma virus(HPV) infection was also as high as 35%(42/120) among pulmonary squamous carcinoma in our previous research. PTEN/MMAC1/TEP1 is the first tumor suppressor with phosphatase activity identified recently. It was found mutated in multiple tumors. In order to study impaction of EBV, HPV and PTEN on genesis of lung cancer : â‘  fragments of EBV and HPV genome as well as sense and antisense PTEN were separately transfected into immortalized human bronchial epithelial cell line TR to observe their respective transformation action; â‘¡loss of heterozygosity(LOH) and inactivation of PTEN in human lung cancers were detected; â‘¢ PTEN was transfected into PTEN absence(PTEN^-/-) small cell lung cancer cell line to observe alteration of cellular growth made by PTEN .1.LMP1 and EBNA2 are the most important transformation related genes of EBV. Eukaryotic expression plasmids pBS-LMP1-Hyg and pSG5-EBNA2-Hyg were cotransfected into TR cell line mediated by Dosper. Hygromycine resistant cell colonies were picked up. In situ hybridization and Western blot were proceeded to confirm the co-expression of LMP1 and EBNA2 on RNA and protein level in TR/LMP1-EBNA2 cell strain. TR/LMP1-EBNA2 cells became smaller and highly reflective. Growth curve and MTT value showed increased growing. Colony formation on soft agar was increased(5.01%,6.82% for TR/LMP1-EBNA2; 0.2%,0.3% for control empty vector cell line;...