Modulation Mechanism Of Inflammatory Mediator By Vitamin D In Prostate Cancer PC3 Cell Line

Prostate cancer(PCa)is the most common non-skin malignancy,It usually develops from prostatic intraepithelial neoplasm to hormone-dependent locally invasive adenocarcinoma,and then to hormone-independent metastatic carcinoma.The natural history and mechanism of this disease is not fully understood.Epidemiological studies have shown that ethnic background,age,genetics,and diet can affect the risk of prostate cancer.Chronic inflammation and other environmental factors including diet may play a role in the development of prostate cancer.More and more evidences show that activated immune cells produce more cytokines and growth factors,which lead to DNA damage and enhance the proliferation and invasiveness of cancer cells.It is the basis of exploring the development and progress of PCa to characterize the role of inflammatory cells in inducing molecular changes.Direct association between some cytokines such as IL-6 and IL-8 and key transcription factors(such as NF-?Bp65?p38),and cancer progression have been demonstrated.Some studies have linked the presence of inflammatory cytokines,chemokines and growth factors to an increased risk of PCa.Inflammatory mediators such as cytokines and chemokines are involved in the specific molecular signal transduction mechanism between immune cells and cancer cells,which can provide theoretical support for the early diagnosis and treatment of PCa.Due to limited knowledge of prostate-specific inflammatory responses,further research is needed to determine the mechanisms by which immune cells infiltrate prostate tissue and their effects on prostate epithelial cells and stromal cells.Therefore,more models are needed to explore the specific role of inflammatory mediators in prostate cancer cells before targeting and pharmacological strategies for single or combination therapy.In addition to bone health,vitamin D,a steroid hormone,also affects a wide range of physiological processes,including inflammation,angiogenesis,apoptosis,differentiation,and cell growth and proliferation.Studies have shown that the risk of PCa is negatively correlated with serum levels of vitamin D,lower calcitriol circulating level is risk factor for prostate cancer,long-term safe doses of vitamin D supplements can prevent or even reverse low-grade prostate cancer.Many recent studies have shown the effect of vitamin D on immune response,but there is still little evidence linking vitamin D to immune response in the context of prostate cancer.To elucidate the mechanism of vitamin D regulating the expression of cytokines and chemokines in inert and invasive PCa may be helpful to understand how vitamin D affects the progress of PCa and to develop better prevention and treatment strategies.Therefore,special attention should be paid to the role of calcitriol and calcitriol in the expression of related factors involved in immune regulation in PCa cells.In order to explore the effect of vitamin D on prostate cancer,this study systematically examined the changes of cell viability of three typical PCa cell lines under different concentrations and different time of two typical vitamin D exposures.In the prostate cancer cell line PC3,the expression of various cytokines and chemokines under different concentrations of vitamin D,and the role of cytokines and chemokines in the tumor microenvironment of PCa in order to elucidate the mechanism of vitamin D regulating the expression of cytokines and chemokines in PCa,and to help develop better prevention and treatment strategies.The results showed that LNCa P cell and DU145 cell activity were maintained above95% at exposure time of 24 hours and 48 hours,respectively,and at 1,10 and 100 n M of calcifediol and calcitriol treatments,indicating the level of vitamin D less than 100 n M had a weak effect on the activity of LNCa P and DU145 cell lines within 48 hours.Under different concentrations of calcitriol and calcifediol,the activity of PC3 cells decreased with the increase of exposure intensity and exposure time.The effect of calcitriol was higher than that of calcifediol.Compared with the LNCa P cell line and the DU145 cell line,the activity of PC3 cell exposed to 100 n M of vitamin D has been reduced to about 90%.The morphology of PC3 cells showed the characteristics of poorly differentiated adenocarcinoma under electron microscopy.Therefore,PC3 cell line was selected as a model for observing the drug action effects.The same exposure concentration and exposure time,i.e.1nM,10 nM and 100 nM of calcitriol and calcitriol,were selected to evaluate their ability to regulate cytokines/chemokines in PC3 cell line after 24 hours and 48 hours exposure.The results indicate that the immunomodulatory effects of vitamin D are dose-dependent.There were no significant differences in the regulation of the expression of multiple inflammatory mediators by calcitriol and calcifediol.Calcitriol and calcifediol have significant effects on IL-1?,IL-6,IL-8,IL-17,TNF-?,TNF-? at three concentration gradients and two time scales of 24 and 48 hours.The significant up-regulation of IL-4,IL-10,and EOTAXIN expression levels have also been shown under vitmamin D treatment.The expression of NF-kappa Bp65 in PC3 cells decreased with the increase of calcifediol exposure concentration at 24 and 48 hours after calcifediol exposure.The expression of NF-kappa Bp65 in PC3 cell decreased significantly at moderate concentration of 10 n M.The inhibition of high concentration of calcifediol at 48 hours was stronger.The inhibitory effect of calcifediol on p38 also depends on the concentration gradient and exhibits a time-regulating effect.Calcitriol has similar effects on PC3 cell,but it has no time-dependent characteristics.This study directly explored the effect of vitamin D on the expression of transcription factors related to the occurrence and development of prostate cancer.In PC3 cell,NF-?Bp65and p38 as the key regulators of cytokine regulation by vitamin D,exhibited an inhibitory effect associated with the exposure strength of calcifediol and calcitriol.Vitamin D regulates inflammatory mediators in prostate cancer cells through important regulatory factors,and calcifediol and calcitriol regulate the expression of inflammatory mediators by inhibting the expression of NF-?Bp65 and p38,which are potentially important cancer preventive factors.It indicates that vitamin D may have a potential prospect in the prevention and treatment of PCa through cytokines.