Investigation Of The Roles Play By Antisense Oligonucleotide For K14 In Keratinocytes

Psoriasis is a autoimmune skin diseases , which is characterized by abnormal proliferation and differentiation of keratinocytes. Due to the complexity of the pathogenesis, psoriasis is not definited totally. It is well known that the interaction between T lymphocytes and the keratinocytes is a very important factor in the pathogenesis of the psoriasis.Some studies found that specific autoantigen actived T lymphocytes consist in psoriatic lesions. The actived T lymphocytes secrete a series of cytokines leading to psoriatic pathologic changes:including keratinocytes hyperplasia , inflammatory cells consisting mainly of T cells infiltration and parakeratin etc.In addition, a number of psoriasis patients are triggred by streptococcal infection firstly, but streptococcal infection is not definitely associated with chronic psoriasis; Some latest studies show that triggering factor maybe streptococcal M protein, which is homologous to keratin 14 peptides in psoriatic lesions. Streptococci may be induce proliferation of various repertoire of T cells(including K14-responsive T cells) possibly through a superantigen-dependent process. Moreover, an epitope on keratin 14 may be a major target for autoreactive T lymphocytes in psoriasis. So Langerhans cell may directly be lined with kerartin throught entering into keratinocytes.Thus,the streptococcal M protein and homologous keratin 14 peptides have been suggested as candidate triggering factors through T lymphocytes activated factors. The recruited M-protein-specific T-cells, which recognize 50 kDa type I keratin(K14) peptide that shares some degree of homology with streptococcal M-protein may be caused and maintained by cross-reactive epitopes in psoriatic lesions. In another study, hyperproliferation epidermal keratinocytes overexpress K14 in psoriatic lesions. Therefore, keratin K14, which recognize T cells as an autoantigen which may be found in psoriasis, is associated with keratinocyte hyperproliferation.This hypothesis,however, has not been clarified yet.The biological role of keratin K14 is still controversial and not well known in humans[6]In order to identify the biological function of the K14 gene and encoding product in the pathogenesis of psoriasis , We show that keratin 14 peptides have been suggested as candidate triggering factors that it is homologous streptococcal M protein. It has therefore been suggested that K14 antisense oligonucleotide can block K14 mRNA and protein expression and inhibit cell proliferation. K14- T lymphocytes may form a cycle interaction pathway and eliminating interaction of K14 protein and streptococcal M-protein, Blocking the cycle can inhibit continued activating of T lymphocytes to achieve acure of psoriasis.It can not be excluded that K14 protein play an important role in the preliferation and function of keratinocytes. Whether the Kl4 as a putative psoriasis autoantigen in vitro directly effect the preliferation and function of keratinocytes? In order to answer such a question, in this study we attempted to look at the possible role of K14 in keratinocytes with antisence technology .The main methods and results are:1 , The identity K14 of Expresion in keratinocytes in vitro As an in vitro model of study in the pathogenesis of psoriasis, we have used the prime culture normal human epidermal keratinocytes. We confirme that this cell in conflune stage exhibits most of the characteristics of basal keratinocytes, including key aspects of K14 expression,and importantly, exhibits the hyperproliferation typical of epidermal keratinocytes in the psoriatic lesion. Immunohistochemistry demonstrated K14 expression in keratinocytes. We confirmed that K14 antisense oligonucleotides transfected confluned keratinocytes, and a higher levels of transfected rate.2, The effect on expression of K14 by liposome conjugated antisense oligonucleotides targeting the mRNA encoding keratin 14 in keratinocytesThe antisense, sense and mismatched oligonucleotides with lipofectin for K14 gene were synthesized and transfected individually to kera...