Expression Of Cytochrome P4502A13Gene In Human Non-small Cell Lung Cancer And Construction Of Lentiviral Vector

Objective:We investigated the expression of Cytochrome P4502A13(CYP2A13)gene in lung cancer and paired normal tissues, then analyzed the relationship between CYP2A13expression and relevant clinical and pathologic data. To construct human cytochrome enzyme P4502A13(CYP2A13) and green fluorescent protein (GFP) co-expressing lentiviral vector, and transfect A549lung cancer cells for highly expressing CYP2A13gene continuously.Methods: Thirty-five lung cancer and paired normal tissues were studied for CYP2A13gene expression using Real-time PCR and Western blot. The correlation between the expression of CYP2A13expression and clinicopathological factors such as age,gender, histology and lymph node status were analyzed. SPSS(17.0)statistical software was applied for data analysis (α=0.05). Gene recombinant technology was employed to link CYP2A13and GFP genes for constructing lentiviral vector. After transfecting293T cells, virus was collected and the titer was measured. The packed lentiviral vector was used to infect A549cells. Then laser scanning confocal microscope (LSCM)was used to observe A549cells and detect the expression of CYP2A13using Real-time PCR and Western blot.Results:The Real-time PCR result showed that there was no significant difference of the CYP2A13mRNA level between tumor and paired normal tissue in the35samples and in12pair squamous cell carcinoma; but in adenocarcinoma, the expression of CYP2A13mRNA in tumor tissues was0.125fold than that in the adjacent tissues (P<0.05).And the expression of CYP2A13mRNA was not associated with the patients’age,gender, histology and lymph node status in tumor. The amounts of CYP2A13proteins revealed by a Western blot analysis correlated well with those of the corresponding mRNAs. A recombinant lentiviral vector co-expressing CYP2A13and GFP gene was constructed successfully. After transfection, a large number of293T cells with green fluorescence were observed under fluorescent microscope. Real-time PCR and Western blotting showed that CYP2A13gene was highly expressed. Conclusions:The expression of CYP2A13was down-regulated in lung adenocarcinoma, and it may play a role of tumor-inhibiting factor. It prompt that CYP2A13maybe involved in the development and progression of lung adenocarcinoma. Successful construction of human CYP2A13and GFP co-expressing lentiviral vector provides a stable vector for further investigation of the relationship between CYP2A13gene and respiratory tumors.