Effect Of Lipoxin A₄ Proliferation And Expression Of IL-6 And VEGF In CSCC A431 Cells

objective: Cutanous squamous cell carcinoma is a malignant tumor,which Originated from the epidermis or adnexal cutin cell.It has a high incidence,malignant degree,a rapid growth and shift.The etiology is not clear.Therefore,studying the cause of disease is particularly important.Lipid oxygen element（Lipoxins LXs）is a kind of lipoxygenase on arachidonic acid metabolites,It could promote inflammation disappeared.in a wide variety of tumor experiments showed antitumor properties.Lipopolysaccharide（lipopolysaccharide,LPS）as a potential cell activation factor,caused by excessive activation of inflammatory cells and inflammatory medium,Inflammation also has close relationship with the occurrence and development of tumor.To explore the antitumor action of LXA₄.We also observe the effects of lipoxin A₄（LXA₄）on the expression of IL-6（interleukin-6）and vascular endothelial growth factor（VEGF）by LPS（lipopolysaccharide）in-duced A₄31 cells.Methods : Using immunohistochemical SP method to detect the expression of ALX levels in CSCC and normal skin tissue.The experiment is divided into five groups: the control group（complete serum medium）,LXA₄（800 n M）group,LPS（1 ug /ml-1）group,LXA₄（800 n M）+ LPS（1 ug/ml-1）group and LXA₄（800 n M）+ LPS（1 ug/ml-1）+ PTX（0.1mg.l-1）group.A₄31 cells were cultured in vitro.The proliferation were assessed by MTT assay,the expression of IL-6 and VEGF was detected by ELISA method,RT-q PCR and Western blot method was used to detect IL-6/VEGF protein and m RNA expression.All the datas by SPSS 17.0 software and graphpad prism 5 software processing andstatistical analysis.Results: 一 、 Immunohistochemical show that ALX is a small amount of expression in normal tissue and is extremely low and even do not express in CSCC.二、MTT experiment results:（1）At the same time,with the increase of drug concentration（LXA₄）,Its inhibition rate also increased,which showed a concentration-dependent;Under the effect of the same drug concentration（LXA₄）,with the extension of time,its inhibition rate also increased,which showed a time-dependent.（2）Different time points determined by MTT experiment results: 24 h,LXA₄ group is lower than the normal control group（P > 0.05）,LPS + LXA₄ group is lower than the normal control group（P >0.05）.48 h,LXA₄ group is lower than the normal control group（P < 0.001）,the LPS group is higher than the normal control group（P < 0.05）,the LPS +LXA₄stimulus is lower than the pure LPS stimulation group（P <0.001）,LPS +LXA₄+ PTX group is higher than the LPS + LXA₄ stimulation group（P <0.001）.72 h,LXA₄ group is lower than the normal control group（P < 0.001）,the LPS group is higher than normal control group（P<0.01）,the LPS + LXA₄ stimulation group is lower than the LPS stimulation group（P < 0.001）,LPS +LXA₄+ PTX group is higher than the LPS + LXA₄ stimulation group（P > 0.05）.三、 ELISA results: LXA₄ group compared with normal control group,IL-6 /VEGF expression is low（P<0.05）;IL-6/VEGF expression of LPS group was higher than normal control group（P<0.05）;IL-6/VEGF expression LPS +LXA₄ group was lower than LPS group（P<0.05）;IL-6/VEGF expressionLXA₄ + LPS+ PTX group was lower than LPS +LXA₄ group（P<0.05）.四、Western blotting and rt-pcr test results: In the LXA₄ group,IL-6/VEGF protein expression and m RNA level were decreased（P >0.05）;In the LPS group,IL-6/ VEGF protein and m RNA levels were increased（P<0.05）;In the LXA₄ + LPS group,IL-6/VEGF protein expression and m RNA level were decreased（P<0.05）;In the LXA₄ + LPS + PTX group,IL-6 /VEGF protein and m RNA expression levels were lower than in the LPS +LXA₄ group（P<0.05）.Conclusion: 1、ALX maybe have a lower expression than normal tissue.2、After LXA₄ stimulus A₄31 cells,A₄31 cells proliferation is restrained,characterized by time-concentration dependent.3、LPS have a certain degree of entrainment to A₄31 cells.4、LXA₄ can inhibit the LPS-induced ex-pression of IL-6/VEGF in A₄31 cells.5、PTX can adersely affect LXA₄ inhibition of LPS.