木犀草素抑制与前列腺癌PC3细胞共培养的内皮细胞增殖、迁移及其分子机制
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摘要
目的探讨木犀草素对与前列腺癌PC3细胞共培养的内皮细胞增殖、迁移的影响及其分子机制。方法原代分离培养人脐静脉内皮细胞(HUVECs)并用VE-钙黏素的表达进行鉴定。使用Transwell系统建立肿瘤-内皮细胞共培养体系。检测不同浓度[0(共培养对照组)、20与50μmol·L~(-1)]木犀草素对共培养系统中HUVECs细胞增殖、迁移的影响。加设不含前列腺癌PC3细胞的内皮细胞对照组。使用人血管新生相关蛋白抗体阵列试剂盒检测各组细胞培养上清液中的蛋白因子水平。结果培养的HUVECs表达VE-钙黏素。与PC3细胞共培养的HUVECs增殖能力比内皮细胞对照组显著增强(P<0.01);以20与50μmol·L~(-1)木犀草素干预可显著抑制共培养体系中HUVECs的增殖能力(均为P<0.01)。与PC3细胞共培养的HUVECs迁移能力比内皮细胞对照组显著增强(P<0.01);以20与50μmol·L~(-1)木犀草素干预可显著抑制共培养体系中HUVECs的迁移能力(均为P<0.01)。共培养体系细胞培养上清液中多种血管新生相关蛋白分泌水平显著高于内皮细胞对照组。以20μmol·L~(-1)木犀草素干预可显著抑制共培养体系细胞培养上清液中白介素-8(IL-8)、血管内皮生长因子(VEGF)及单核细胞趋化蛋白-1(MCP-1)的分泌水平(均为P<0.01)。结论木犀草素可能通过抑制肿瘤-内皮细胞共培养体系中IL-8、VEGF及MCP-1的分泌抑制内皮细胞的增殖与迁移。
OBJECTIVE To explore the effects of luteolin on the proliferation and migration of the endothelial cells co-cultured with cancer cells and its molecular mechanism. METHODS Human umbilical vein endothelial cells( HUVECs) were primarily isolated and identified by the expression of VE-cadherin. Cancer-endothelial cell co-culture model was established using the Transwell system. The effects of luteolin at different concentrations( 0 [Co-culture control],20 and 50 μmol·L~(-1)) on the proliferation and migration of HUVECs in the co-culture system were determined. A HUVECs control group removed of prostate cancer PC3 cells was also included. Human angiogenesis antibody array kit was used to assay the secretion levels of various protein factors in each group. RESULTS VE-cadherin was expressed on all the cultured HUVECs. Increased proliferation ability was found in the HUVECs co-cultured with PC3 cells compared with that in HUVECs control group( P < 0. 01). Treatment with 20 or 50 μmol·L~(-1) luteolin significantly inhibited the proliferation ability of the HUVECs in co-culture system( both P < 0. 01). Increased migration ability was found in the HUVECs co-cultured with PC3 cells compared with that in HUVECs control group( P < 0. 01). Treatment with 20 or 50 μmol·L~(-1) luteolin significantly inhibited the migration ability of the HUVECs in co-culture system( both P < 0. 01). Secretion levels of multiple angiogenesis-related proteins in the cultural supernatant of co-culture system were significantly increased compared with those in HUVECs control group. Treatment with 20 μmol·L~(-1) luteolin significantly inhibited the secretion levels of interleukin-8( IL-8),vascular endothelial growth factor( VEGF) and monocyte chemoattractant protein-1( MCP-1) in the cultural supernatant of co-culture system( all P < 0. 01). CONCLUSION Luteolin may inhibit the proliferation and migration of endothelial cells via suppressing the secretions of IL-8,VEGF and MCP-1 in cancer-endothelial co-culture system.
OBJECTIVE To explore the effects of luteolin on the proliferation and migration of the endothelial cells co-cultured with cancer cells and its molecular mechanism. METHODS Human umbilical vein endothelial cells( HUVECs) were primarily isolated and identified by the expression of VE-cadherin. Cancer-endothelial cell co-culture model was established using the Transwell system. The effects of luteolin at different concentrations( 0 [Co-culture control],20 and 50 μmol·L~(-1)) on the proliferation and migration of HUVECs in the co-culture system were determined. A HUVECs control group removed of prostate cancer PC3 cells was also included. Human angiogenesis antibody array kit was used to assay the secretion levels of various protein factors in each group. RESULTS VE-cadherin was expressed on all the cultured HUVECs. Increased proliferation ability was found in the HUVECs co-cultured with PC3 cells compared with that in HUVECs control group( P < 0. 01). Treatment with 20 or 50 μmol·L~(-1) luteolin significantly inhibited the proliferation ability of the HUVECs in co-culture system( both P < 0. 01). Increased migration ability was found in the HUVECs co-cultured with PC3 cells compared with that in HUVECs control group( P < 0. 01). Treatment with 20 or 50 μmol·L~(-1) luteolin significantly inhibited the migration ability of the HUVECs in co-culture system( both P < 0. 01). Secretion levels of multiple angiogenesis-related proteins in the cultural supernatant of co-culture system were significantly increased compared with those in HUVECs control group. Treatment with 20 μmol·L~(-1) luteolin significantly inhibited the secretion levels of interleukin-8( IL-8),vascular endothelial growth factor( VEGF) and monocyte chemoattractant protein-1( MCP-1) in the cultural supernatant of co-culture system( all P < 0. 01). CONCLUSION Luteolin may inhibit the proliferation and migration of endothelial cells via suppressing the secretions of IL-8,VEGF and MCP-1 in cancer-endothelial co-culture system.
引文
[1]ZHANG J X,XING J G,WANG L L,et al.Luteolin inhibits fibrillary beta-amyloid1-40-induced inflammation in a human blood-brain barrier model by suppressing the p38 mapk-mediated nf-kappab signaling pathways[J].Molecules,2017,22(3):334-336.
[2]LIU Y,HUANG J,ZHENG X,et al.Luteolin,a natural flavonoid,inhibits methylglyoxal induced apoptosis via the mtor/4ebp1 signaling pathway[J].Sci Rep,2017,7(1):7877-7879.
[3]ZANG M D,HU L,FAN Z Y,et al.Luteolin suppresses gastric cancer progression by reversing epithelial-mesenchymal transition via suppression of the notch signaling pathway[J].J Transl Med,2017,15(1):52-53.
[4]KANG K A,PIAO M J,RYU Y S,et al.Luteolin induces apoptotic cell death via antioxidant activity in human colon cancer cells[J].Int J Oncol,2017,51(4):1169-1178.
[5]ZANG M,HU L,ZHANG B,et al.Luteolin suppresses angiogenesis and vasculogenic mimicry formation through inhibiting notch1-vegf signaling in gastric cancer[J].Biochem Biophys Res Commun,2017,490(3):913-919.
[6]GEORGE V C,DELLAIRE G,RUPASINGHE H P V.Plant flavonoids in cancer chemoprevention:role in genome stability[J].J Nutr Biochem,2017,45(1):1-14.
[7]BAUDIN B,BRUNEEL A,BOSSELUT N,et al.A protocol for isolation and culture of human umbilical vein endothelial cells[J].Nat Protoc,2007,2(3):481-485.
[8]CHIEW G G,FU A,LOW K P,et al.Physical supports from liver cancer cells are essential for differentiation and remodeling of endothelial cells in a Hep G2-HUVEC co-culture model[J].Sci Rep,2015,5(1):1080-1082.
[9]HUSS W J,HANRAHAN C F,BARRIOS R J,et al.Angiogenesis and prostate cancer:identification of a molecular progression switch[J].Cancer Res,2001,61(6):2736-2743.
[10]PAYTON S.Prostate cancer:galectin signature reveals gal-1 as key player in angiogenesis[J].Nat Rev Urol,2012,9(12):667-671.
[11]MIYATA Y,OHBA K,MATSUO T,et al.Tumor-associated stromal cells expressing e-prostanoid 2 or 3 receptors in prostate cancer:correlation with tumor aggressiveness and outcome by angiogenesis and lymphangiogenesis[J].Urology,2013,81(1):136-142.
[12]FERRANDO M,GUERON G,ELGUERO B,et al.Heme oxygenase 1(ho-1)challenges the angiogenic switch in prostate cancer[J].Angiogenesis,2011,14(4):467-479.
[13]BAGLI E,STEFANIOTOU M,MORBIDELLI L,et al.Luteolin inhibits vascular endothelial growth factor-induced angiogenesis;inhibition of endothelial cell survival and proliferation by targeting phosphatidylinositol 3'-kinase activity[J].Cancer Res,2004,64(21):7936-7946.
[14]SUI J Q,XIE K P,XIE M J.Inhibitory effect of luteolin on the proliferation of human breast cancer cell lines induced by epidermal growth factor[J].Acta Physiol Sin(生理学报),2016,68(1):27-34.
[15]CAI X,LU W,YE T,et al.The molecular mechanism of luteolin-induced apoptosis is potentially related to inhibition of angiogenesis in human pancreatic carcinoma cells[J].Oncol Rep,2012,28(4):1353-1361.
[16]KIM J A,KIM D K,KANG O H,et al.Inhibitory effect of luteolin on tnf-alpha-induced il-8 production in human colon epithelial cells[J].Int Immunopharmacol,2005,5(1):209-217.
[17]LI W F,ZHOU K,ZHAO Z B.Inhibition effect of luteolin on IL-8 signal path inbreast cancer cells MDA MB 231[J].Chin J Adv Gen Sure(中国现代普通外科进展),2014,17(6):435-438.
[18]DING H,LI D,ZHANG Y,et al.Luteolin inhibits smooth muscle cell migration and proliferation by attenuating the production of nox4,p-akt and vegf in endothelial cells[J].Curr Pharm Biotechnol,2014,14(12):1009-1015.
[19]WU S H,LOU Y,HUANG L.Progress of in the mechanism of monocyte chemoattractant protein-1[J].Chin J Clin(Elect Ed)(中华临床医师杂志电子版),2012,6(10):2773-2775.
[20]BHARDWAJ S,ROY H,BABU M,et al.Adventitial gene transfer of vegfr-2 specific vegf-e chimera induces mcp-1 expression in vascular smooth muscle cells and enhances neointimal formation[J].Atherosclerosis,2011,219(1):84-91.
[2]LIU Y,HUANG J,ZHENG X,et al.Luteolin,a natural flavonoid,inhibits methylglyoxal induced apoptosis via the mtor/4ebp1 signaling pathway[J].Sci Rep,2017,7(1):7877-7879.
[3]ZANG M D,HU L,FAN Z Y,et al.Luteolin suppresses gastric cancer progression by reversing epithelial-mesenchymal transition via suppression of the notch signaling pathway[J].J Transl Med,2017,15(1):52-53.
[4]KANG K A,PIAO M J,RYU Y S,et al.Luteolin induces apoptotic cell death via antioxidant activity in human colon cancer cells[J].Int J Oncol,2017,51(4):1169-1178.
[5]ZANG M,HU L,ZHANG B,et al.Luteolin suppresses angiogenesis and vasculogenic mimicry formation through inhibiting notch1-vegf signaling in gastric cancer[J].Biochem Biophys Res Commun,2017,490(3):913-919.
[6]GEORGE V C,DELLAIRE G,RUPASINGHE H P V.Plant flavonoids in cancer chemoprevention:role in genome stability[J].J Nutr Biochem,2017,45(1):1-14.
[7]BAUDIN B,BRUNEEL A,BOSSELUT N,et al.A protocol for isolation and culture of human umbilical vein endothelial cells[J].Nat Protoc,2007,2(3):481-485.
[8]CHIEW G G,FU A,LOW K P,et al.Physical supports from liver cancer cells are essential for differentiation and remodeling of endothelial cells in a Hep G2-HUVEC co-culture model[J].Sci Rep,2015,5(1):1080-1082.
[9]HUSS W J,HANRAHAN C F,BARRIOS R J,et al.Angiogenesis and prostate cancer:identification of a molecular progression switch[J].Cancer Res,2001,61(6):2736-2743.
[10]PAYTON S.Prostate cancer:galectin signature reveals gal-1 as key player in angiogenesis[J].Nat Rev Urol,2012,9(12):667-671.
[11]MIYATA Y,OHBA K,MATSUO T,et al.Tumor-associated stromal cells expressing e-prostanoid 2 or 3 receptors in prostate cancer:correlation with tumor aggressiveness and outcome by angiogenesis and lymphangiogenesis[J].Urology,2013,81(1):136-142.
[12]FERRANDO M,GUERON G,ELGUERO B,et al.Heme oxygenase 1(ho-1)challenges the angiogenic switch in prostate cancer[J].Angiogenesis,2011,14(4):467-479.
[13]BAGLI E,STEFANIOTOU M,MORBIDELLI L,et al.Luteolin inhibits vascular endothelial growth factor-induced angiogenesis;inhibition of endothelial cell survival and proliferation by targeting phosphatidylinositol 3'-kinase activity[J].Cancer Res,2004,64(21):7936-7946.
[14]SUI J Q,XIE K P,XIE M J.Inhibitory effect of luteolin on the proliferation of human breast cancer cell lines induced by epidermal growth factor[J].Acta Physiol Sin(生理学报),2016,68(1):27-34.
[15]CAI X,LU W,YE T,et al.The molecular mechanism of luteolin-induced apoptosis is potentially related to inhibition of angiogenesis in human pancreatic carcinoma cells[J].Oncol Rep,2012,28(4):1353-1361.
[16]KIM J A,KIM D K,KANG O H,et al.Inhibitory effect of luteolin on tnf-alpha-induced il-8 production in human colon epithelial cells[J].Int Immunopharmacol,2005,5(1):209-217.
[17]LI W F,ZHOU K,ZHAO Z B.Inhibition effect of luteolin on IL-8 signal path inbreast cancer cells MDA MB 231[J].Chin J Adv Gen Sure(中国现代普通外科进展),2014,17(6):435-438.
[18]DING H,LI D,ZHANG Y,et al.Luteolin inhibits smooth muscle cell migration and proliferation by attenuating the production of nox4,p-akt and vegf in endothelial cells[J].Curr Pharm Biotechnol,2014,14(12):1009-1015.
[19]WU S H,LOU Y,HUANG L.Progress of in the mechanism of monocyte chemoattractant protein-1[J].Chin J Clin(Elect Ed)(中华临床医师杂志电子版),2012,6(10):2773-2775.
[20]BHARDWAJ S,ROY H,BABU M,et al.Adventitial gene transfer of vegfr-2 specific vegf-e chimera induces mcp-1 expression in vascular smooth muscle cells and enhances neointimal formation[J].Atherosclerosis,2011,219(1):84-91.