A Pilot Study On Lipoxin A₄ And Transforming Growth Factor Beta 1 In Malignant Melanoma

objective:Malignant melanoma?MM?is one of the cancer with highest death rate,and there is no effective treatment for advanced MM.Lipoxins?LXs?is a kind of metabolic product of arachidonic acid,which can promote the regression of inflammation,and it has the characteristics of anti-tumor in many researches.Transforming growth factor beta 1?TGF-?1?is a multifunctional regulating growth factor involved in the regulation of cell growth and differentiation,and it has a close relationship with the occurrence and development of tumor.The rapidly developing fields of melanoma research are revolutionizing the current concepts on melanoma etiology and pathogenesis and are introducing potential therapeutic approaches.This study aimed to investigate the role of Lipoxin A₄?LXA₄?and TGF-?1 in the occurrence and development of malignant melanoma in vitro.Methods:The expression level of ALX and TGF-?1 were studied in normal skin tissues and skin lesions of the malignant melanoma patients with SP immunohistochemical technique.Malignant melanoma cells A375 were randomly divided into four groups,and complete medium?LXA₄ factor?400nM??TGF-?1 factor?50ng/ml?and both of them were respectively added into each group.After stimulation,the cell ability of proliferation and invasion and the level of apoptosis were respectively detected by CCK-8 cell proliferation experiment?Transwell tumor invasion assay and apoptosis experiment of flow cytometry.Meanwhile,the expressions of ALX and TGF-?1 in A375 cells were detected by Western Blot experiment and qRT-PCR experiment from the levels of translation and transcription.Finally,all the experimental results were analyzed by using SPSS17.0 software.Results:The IHC analysis of malignant melanomas and normal skin tissues revealed ALX and TGF-?1 to be predominantly expressed by melanoma cells?p<0.05?.The results of CCK-8 cell proliferation assay showed that the survival rates of the normal control group at each point of time were higher than that in the other experimental groups?p<0.05?.In Transwell tumor cell invasion assay,the number of invasive cells in the TGF-?1 factor group was higher than that in the normal control group,while that of the LXA₄ factor group was lower than that of the normal control group?p<0.05?.The results of flow cytometry apoptosis experiment showed that the number of apoptotic cells in LXA₄ factor group was much more than that in the normal control group?p<0.001?.The results of Western Blot experiment and qRT-PCR experiment showed that,the expression levels of translation and transcription of ALX in LXA₄ factor group were higher than that in the normal control group?p<0.05?,at the same time,the expression levels of translation and transcription of TGF-?1 in LXA₄ factor group were lower than that in the normal control group?p<0.05?;however,the transcription and translation levels of ALX in TGF-?1factor group were decreased compared with the normal control group?p>0.05?,moreover,the transcription and translation levels of TGF-?1 in TGF-?1 factor group were higher than those in the normal control group?p<0.05?.Conclusion:LXA₄ and TGF-?1 had a certain correlation with the occurrence and development of malignant melanoma;LXA₄,to a certain extent,inhibited the proliferation?invasion and metastasis capabilities of malignant melanoma cells,and promoted apoptosis of MM cells;TGF-?1 showed a double effects on MM cells,which inhibited proliferation in early stage and promoted invasion and metastasis in advanced stage;in addition,between LXA₄ and TGF-?1,there is a certain degree of mutual antagonism.