Effect Of Autophagy In Cancer Associated Fibroblasts On The Progression Of Prostate Cancer Cell

Background: As a worldwide issue,Prostate Cancer(PCa)seriously endangers the health of the aging male,and is also the main cause of death in aging male.However,the specific mechanism of PCa formation and progression remains unclear.Recently,treatment of targeting tumor microenvironment is in the limelight in cancer.Fibroblasts,the primary mesenchymal cells in tumor microenvironment,was caused great attention.We found that Cancer-associated fibroblasts(CAF)demonstrate high level of basal autophagy in prostate cancer compared with normal fibroblast.Furthermore,autophagy was thought to play a big role in tumorigenesis and tumor progression.Hence,it may be a promising cure to target CAF,especially autophagy in CAF,in numerous diseases.Here,we hypothesized that CAF autophagy can promote PCa progression.We devised an experiment in which the level of autophagy in CAF contributes to the progression of prostate cancer cells.Therefore,this study will provide a new theoretical support and treatment strategies for the study and treatment of prostate cancer.Methods and Materials: First,the autophagy levels of CAF were detected by immunoblotting.Then,the autophagy inhibitor chloroquine was used to inhibit autophagy of CAF.In order to further confirm whether the autophagy level of CAF is inhibited,the marker protein LC-3Ⅱ that represents autophagy levels was tested by immunoblotting.And then co-culture system of CAF and prostate cancer cells was established,and proliferation and invasion capabilities of prostate cancer cells were detected by CCK-8 assay and Transwell.Meanwhile,the autophagy level of CAF was up-regulated by upregulating the autophagy initiation gene Beclin-1,and we confirm that levels of Beclin-1 and autophagy in CAF were up-regulated by immunoblotting,and then Co-culture system of CAF and prostate cancer cells was established,and proliferation and invasion capabilities of prostate cancer cells were detected by CCK-8 assay and Transwell.Finally,we detected the levels of related proteins(MMP-9,MMP-2)that represent the invasion ability of prostate cancer cells by immunoblotting.Results: Immunoblot results showed that the level of LC3-Ⅱ in CAF was higher than NF,so CAF had a higher autophagy level than NF.After inhibiting CAF autophagy,coculture system of CAF and prostate cancer cells was established.The results of CCK-8 and Transwell showed the ability of proliferation and invasion of prostate cancer cell is significantly reduced.After up-regulating the autophagy initiation gene Beclin-1 in CAF by viral vectors,immunoblot results confirmed that the expression of Beclin-1 was upregulated,and the CAF autophagy level was also up-regulated.After co-culture,the ability of proliferation and invasion of prostate cancer cells was enhanced.Immunoblotting results showed that the expression levels of related proteins(MMP-9,MMP-2)that represent invasion ability in prostate cancer cells were increased.Conclusion: The level of autophagy in CAF was higher than NF.Autophagy of cancer-associated fibroblasts promotes prostate cancer cell progression.