[FHL1 knockdown mediated by lentiviral shRNA promotes the growth of HeLa and HepG2 cells].

OBJECTIVE

To investigate the effect of four-and-a-half LIM domain 1 (FHL1) knockdown by lentiviral-mediated shRNA on the growth of HeLa and HepG2 cells.

METHODS

pLenti-H1 FHL1 shRNA was cloned, and then transfered into HEK293T cells. The inhibitory effect of pLenti-H1 FHL1 shRNA on FHL1 gene was detected by Western blotting and real-time quantitative PCR (qRT-PCR). Lentivirus particles were packaged, added to HeLa and HepG2 cells, followed by puromycin treatment for 2-3 weeks to screen stable clones. The knockdown effect on FHL1 expression in these cells was checked by Western blotting and qRT-PCR. Cell growth and colony formation analysis were performed to investigate the effect of FHL1 down-regulation on tumor cell growth. Soft agar analysis was used to analyze its effect on anchorage-independent tumor cell growth.

RESULTS

Western blotting and qRT-PCR revealed that the pLenti-H1 FHL1 shRNA apparently inhibited the expression of FHL1 gene. Cell growth and colony formation assay showed that the lentiviral-mediated shRNA for FHL1 gene significantly accelerated the tumor cell growth in HepG2 and HeLa cells. Soft agar analysis demonstrated that FHL1 shRNA increased the anchorage-independent growth of tumor cells.

CONCLUSION

pLenti-H1 FHL1 shRNA could significantly accelerate tumor cell growth via inhibiting the expression of FHL1.