The Exploration Of Mechanism Of Low Dose Chromium Exposure Promoting Proliferation Of Prostate Cancer Cells

Objective: To explore the possible mechanism of low concentration chromium exposure promoting prostate cancer cell growth,explore the role and mechanism of CEBPB in chromium-stimulated prostate cancer cell growth through IL6/ STAT3 signaling pathway,and explore the role and mechanism of TGF-beta-EMT signaling pathway in low concentration chromium exposure promoting prostate cancer progression.Methods:1)PC3 was exposed to low concentration of chromium.The growth of PC3 cells exposed to low concentration of chromium was detected by MTT and other methods.Total RNA and total protein were collected.The expression level of CEBPB mRNA was detected by real-time fluorescence quantitative PCR,and the expression level of CEBPB protein was detected by WB assay.SiRNA targeting CEBPB was synthesized and transfected into prostate cancer PC3 cell line by Lipofectamine3000.The expression of CEBPB mRNA was detected by real-time fluorescence quantitative PCR.The growth ability of PC3 cell was detected by MTT assay.2)Total RNA,protein and total protein were collected and real-time detected.The mRNA expression of IL6 was detected by fluorescence quantitative PCR,the protein expression of IL6 was detected by ELISA and WB,and the phosphorylated STAT3 and total STAT3 were detected by WB.In order to explore the role of IL6/ STAT3 signaling pathway in this process and inhibit the activation of STAT3 by small molecule inhibitors,MTT assay was used to detect the changes of PC3 cell growth ability after IL6/ STAT3 signaling pathway was inhibited;3)WB was used to detect the level of TGF-beta in PC3 cells exposed to low concentration of IL6/ STAT3,and to speculate that it could promote prostate cancer cells.TGF-beta may play a role in the invasion process in vitro,and explore whether TGF-beta can activate the EMT pathway by detecting the expression of related factors of EMT pathway.After specific reduction of TGF-beta siRNA levels,the expression of EMT pathway-related factors was determined to determine whether the abnormal expression of TGF-beta siRNA could be reversed.Results:1)it was found that low concentration of chromium exposure could significantly promote the growth of prostate cancer cell PC3.The results showed that low concentration of chromium exposure could significantly up-regulate the expression of CEBPB RNA and protein in prostate cancer cell PC3.After knocking down CEBPB expression with siRNA,the growth of prostate cancer cell PC3 was significantly inhibited by low concentration of chromium exposure.2)The results showed that low concentration of chromium exposure could significantly up-regulate the expression of IL-6 mRNA in prostate cancer cell PC3 and up-regulate the secretion of IL-6 and the expression of total protein in prostate cancer cell PC3.Ping significantly increased the level of phosphorylated STAT3.Inhibition of STAT3 activity significantly inhibited the growth of prostate cancer cell PC3 induced by low concentration of chromium exposure.After knocking down CEBPB expression with siRNA,IL6 mRNA and protein expression were down-regulated,phosphorylated STAT3 levels were down-regulated,and IL6/ STAT3 signaling pathway was inhibited.3)The results showed that TGF-beta levels in PC3 cells exposed to low concentrations increased significantly,and the expression of EMT pathway-related factors increased significantly.When TGF beta siRNA was specifically lowered,the abnormal expression of related factors in EMT pathway was reversed.Conclusion:1)Low concentration of chromium exposure can significantly promote the growth of prostate cancer cells PC 3;2)Low concentration of chromium exposure can promote the invasion of prostate cancer cells by up-regulating the expression of CEBPB and activating the IL6/ STAT3 signaling pathway in PC3;3)Low concentration of chromium exposure may promote the invasion of prostate cancer cells through TGF-?-EMT signal.Pathways are promoting the progression of prostate cancer.