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AIM:To investigate the effect of oridonin on nuclear transcription factors and to study the relationship between biological behavior and inflammatory factors in human pancreatic cancer(BxPC-3)cells.METHODS:BxPC-3 cells were treated with various concentrations of oridonin,and viability curves were generated to test for inhibitory effects of the drug on cells.The expression of cytokines such as interleukin-1β(IL-1β),IL-6,or IL-33 was detected in BxPC-3 cell supernatants using an enzyme-linked immunosorbent assay(ELISA),and the protein expression of nuclear transcription factors including nuclear factorκB,activating protein-1,signal transducer and activator of transcription 3,bone morphogenetic protein 2,trans-forming growth factorβ1 and sma and mad homologues in BxPC-3 cells was detected using Western blot.Carcinoma hallmark-related proteins such as survivin,vascular endothelial growth factor,and matrix metallopeptidase 2 were also detected using immunoblotting,and intra-nuclear IL-33 expression was detected using immunofluorescent staining.RESULTS:Treatment with oridonin reduced the viability of BxPC-3 cells in a dose dependent manner.The cells exhibited reduced growth following treatment with 8μg/mL oridonin(13.05%±3.21%,P<0.01),and the highest inhibitory ratio was 90.64%±0.70%,which was achieved with oridonin at a dose of32μg/mL.The IC50 value of oridonin in BxPC-3 cells was 19.32μg/mL.ELISA analysis revealed that oridonin down-regulated the inflammatory factors IL-1β,IL-6,and IL-33 in a dose-dependent manner.IL-1βexpression was significantly reduced in the 16 and 32μg/mL treatment groups compared to the control group(12.97±0.45 pg/mL,11.17±0.63 pg/mL vs 14.40±0.38pg/mL,P<0.01).Similar trends were observed for IL-6expression,which was significantly reduced in the 16and 32μg/mL treatment groups compared to the control group(4.05±0.14 pg/mL vs 4.45±0.43 pg/mL,P<0.05;3.95±0.13 pg/mL vs 4.45±0.43 pg/mL,P<0.01).IL-33 expression was significantly reduced in the8,16,and 32μg/mL treatment groups compared to the control group(911.05±14.18 pg/mL vs 945.25±12.09 pg/mL,P<0.05;802.70±11.88 pg/mL,768.54±10.98 pg/mL vs 945.25±12.09 pg/mL,P<0.01).Western blot and immunofluorescent staining analyses suggested that oridonin changed the hallmarks and regulated the expression of various nuclear transcription factors.CONCLUSION:The results obtained suggest that oridonin alters the hallmarks of pancreatic cancer cells through the regulation of nuclear transcription factors.
AIM: To investigate the effect of oridonin on nuclear transcription factors and to study the relationship between biological behavior and inflammatory factors in human pancreatic cancer (BxPC-3) cells. METHODS: BxPC-3 cells were treated with various concentrations of oridonin, and viability ups were generated to test for inhibitory effects of the drug on cells. The expression of cytokines such as interleukin-1β (IL-1β), IL-6, or IL-33 was detected in BxPC-3 cell supernatants using an enzyme-linked immunosorbent assay (ELISA), and the protein expression of nuclear transcription factors including nuclear factor κB, activating protein-1, signal transducer and activator of transcription 3, bone morphogenetic protein 2, trans-forming growth factor β1 and sma and mad homologues in BxPC-3 cells were detected using Western blot. Carcinoma hallmark-related proteins such as survivin, vascular endothelial growth factor, and matrix metallopeptidase 2 were also detected using immunoblotting, and intra-nuclear IL-3 3 expression was detected using immunofluorescent staining. RESULTS: Treatment with oridonin reduced the viability of BxPC-3 cells in a dose dependent manner. These cells exhibited reduced growth following treatment with 8 μg / mL oridonin (13.05% ± 3.21%, P <0.01) , and the highest inhibitory ratio was 90.64% ± 0.70%, which was achieved with oridonin at a dose of 32 μg / mL. IC50 value of oridonin in BxPC-3 cells was 19.32 μg / mL. ELISA analysis revealed that oridonin down-regulated the inflammatory factors IL-1β, IL-6, and IL-33 in a dose-dependent manner. IL-1βexpression was significantly reduced in the 16 and 32 μg / mL treatment groups compared to the control group (12.97 ± 0.45 pg / mL, ± 0.63 pg / mL vs. 14.40 ± 0.38 pg / mL, P <0.01). Similar trends were observed for IL-6 expression, which was significantly reduced in the 16 and 32 μg / mL treatment groups compared to the control group (4.05 ± 0.14 pg / mL vs 4.45 ± 0.43 pg / mL, P <0.05; 3.95 ± 0.13 pg / mL vs. 4.45 ± 0.43 pg / mL, P <0.01) ced in the8,16, and32 μg / mL treatment groups compared to the control group (911.05 ± 14.18 pg / mL vs 945.25 ± 12.09 pg / mL, P <0.05; 802.70 ± 11.88 pg / mL, 768.54 ± 10.98 pg / mL vs 945.25 ± 12.09 pg / P <0.01). Western blot and immunofluorescent staining analysis suggested that oridonin changed the hallmarks and regulated the expression of various nuclear transcription factors. CONCLUSION: The results obtained suggest that oridonin alters the hallmarks of pancreatic cancer cells through the regulation of nuclear transcription factors.