[Proliferation inhibition of human lung adenocarcinoma cell line A549 transfected by RASSF1A gene].

OBJECTIVE

To determine the effects of exogenous RASSF1A gene on the proliferation and expression of P65 and subunit of NF-kappaB, in lung adenocarcinoma cell line A549.

METHODS

pcDNA3.0-RASSF1A and pcDNA3. 0 were introduced into A549 cell line by lipofectin transfection, and the A549 cells stably expressing RASSF1A gene were established by G418 selection. The expression of RASSF1A was detected by Western blotting. The cytobiologic characterizations of the positive clone were analyzed by methythiazoletertraolium (MTT) assay and cytometry. The expressing of P65 was analyzed by RT-PCR and Western blotting.

RESULTS

A549 cells stably expressing RASSF1A protein were established by lipofection mediated transfection and selected for further study. Compared with the nontransfected and vector transfected cells, the positive clone cells grew more slowly. Flow cytometric data showed that more positive clone cells went into phase G0/G1 and fewer cells went into phase S. The expression of P65 in nuclear protein in positive clone cells was lower than that of the control group while there was no obvious difference between the expression of p65 mRNA and P65 protein in total protein among the 3 groups.

CONCLUSION

RASSF1A gene might suppress the proliferation of A549 cells through blocking the activity of P65 protein.