Effects of K17 antisense oligonucleotides on proliferation and expression of K17 in keratinocytes

Objective To investigate the effects of antisense oligonucleotides of keratin 17 (K17) on the proliferation and expression of K17 in keratinocytes in vitro. Methods The antisense, sense and mismatched oligonucleotides for K17 gene were synthesized and conjugated with lipofectin, respectively. They were subsequently transfected into keratinocyte cell line HaCaT in vitro. Cell proliferation was detected with MTT colorimetric assay. K17 mRNA level was detected by reverse transcriptase polymerase chain reaction (RT-PCR).K17 protein level was detected by Western blot, immuno-fluorescent cytochemistry combined with laser scanning confocal microscopy (LSCM). Results The proliferation of HaCaT cells was inhibited and the expression of K17 gene and protein was markedly decreased after the treatment with antisense rather than with sense and mismatched K17 oligonueleotides. Conclusion Antisense oligonucleotides conjugated with lipofectin might be a useful method to inhibit the proliferation of keratinocytes by inhibiting the expression of K17 mRNA and protein.