| Objective: Through the detection of the expression of Non-Small Cell Lung Cancer ’s tumor suppressor-gene TCF21, explored its relationship with the occurrence, development and clinical feature of NSCLC. Then explored the feasibility and validity of gene TCF21 in treating lung cancer by using molecular biology methods such as genetic recombination.Methods: Non selective into groups of 206 lung cancer patients as experimental group,and 60 cases of normal lung tissue as control group;By immunohistochemistry(SP method) to detect TCF21 expression in tissues and observed their significance on clinical pathological characteristic in NSCLC;Then the recombinant advenovirus plasmid p Ad-TCF21 was constructed by homologous recombination in bacteria. Preparation of recombinant TCF21 adenovirus by adenovirus packing experiment; Take the recombinant adenovirus of TCF21 transfer to A549 cells. Detection the expression in A549 cells by fluorescence microscope and Real-Time PCR;To examined the cell proliferation by MTT method,and used cell migration experiment to examined the cell invasion ability.Results: The lower expression of TCF21 was observed in NSCLC(P<0.05);The expression of TCF21 was correlated with clinical stage,degree of tumor differentiation and lymph node metastasis(P<0.05); The recombinant adenoviral plasmid containing TCF21(p Ad-TCF21)was successfully constructed after gene sequencing. The recombined TCF21 adenovirus has been developed successfully. The virus titer is 5.6×1010pfu/ml, which is consistent with the experiment requirements. After the recombined TCF21 adenovirus(Ad-TCF21) infects the A549 cell, the real-Time PCR test indicated that the target gene protein has been expressed successfully in the A549 gene and that the proliferation & invasion ability of the A549 gene has been restrained after the TCF21 gene is expressed. Conclusion: The TCF21 gene plays an important role in generation and growth of NSCLC, and itsfunctional loss may be one of the important mechanisms for generation & growth of the lung cancer. In this experiment, the recombined TCF21 adenovirus has been developed successfully, and has been proved to have the remarkable inhibition effect on proliferation and metastasis ability of the lung cancer cells, which lays certain foundation for further study of the gene therapy of lung cancer. |
