1,25-(OH)_2VitD_3对人胰腺癌细胞侵袭迁移能力及MMP-2、9、14表达的影响
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摘要
目的观察1,25-二羟基维生素D3[1,25-(OH)_2VitD_3]对人胰腺癌PANC-1细胞侵袭迁移能力的影响,以及基质金属蛋白酶(MMP)-2、9、14的表达变化。方法将不同浓度(0、25、50、75、100μmol/L)的1,25-(OH)_2VitD_3作用于PANC-1细胞,干预24 h时分别用Transwell细胞侵袭、迁移实验观察细胞的侵袭和迁移能力,干预48 h时用Western blot法检测细胞MMP-2、9、14蛋白。结果 0、25、50、75、100μmol/L的1,25-(OH)_2VitD_3干预PANC-1细胞24 h时,侵袭细胞数分别为(46.20±0.84)、(30.80±2.28)、(18.80±1.64)、(13.20±2.39)、(5.20±1.30)个/HP,迁移细胞数分别为(66.03±1.25)、(55.82±1.65)、(41.43±1.51)、(19.86±1.45)、(10.23±1.95)个/HP;随着1,25-(OH)_2VitD_3浓度增加侵袭及迁移细胞数逐渐减少,差异有统计学意义(P均<0.05)。1,25-(OH)_2VitD_3干预PANC-1细胞48 h后,随着1,25-(OH)_2VitD_3浓度增加PANC-1细胞MMP-2、9、14蛋白相对表达量逐渐减少,差异有统计学意义(P均<0.05)。结论 1,25-(OH)_2VitD_3可通过下调MMP-2、9、14蛋白的表达,从而抑制胰腺癌PANC-1细胞的侵袭与迁移。
Objective To investigate the effect of 1,25-dihydroxyvitamin D_3[1,25(OH)_2D_3]on invasive and migratory abilities of human pancreatic cancer cell line PANC-1 and the changes in the expression of matrix metalloproteinases( MMP)-2,9 and 14. Methods The routinely cultured human pancreatic cancer cell line PANC-1 in vitro was treated with different concentrations of 1,25( OH)_2D_3( 0,25,50,75 and 100 μmol/L). After 24 hours,the abilities of migration and invasion in vitro in each group were detected by Transwell migration and invasion assays. After 48 hours,the protein expression of MMP-2,9 and 14 in each group was measured by Western blotting. Results After intervention for 24 h with different concentrations of 1,25( OH)_2D_3( 0,25,50,75 and 100 μmol/L),every field's invasive cells in each group were( 46. 20 ± 0. 84),( 30. 80 ± 2. 28),( 18. 80 ± 1. 64),( 13. 20 ± 2. 39) and( 5. 20 ± 1. 30) / HP; and the migratory cells in each group were( 66. 03 ± 1. 25),( 55. 82 ± 1. 65),( 41. 43 ± 1. 51),( 19. 86 ± 1. 45) and( 10. 23 ± 1. 95) /HP,respectively. The migratory and invasive cells were decreased with the increase of concentration,and the difference was statistically significant( all P < 0. 05). After intervention for 48 h with different concentrations of 1,25( OH)2D_3,MMP-2,9 and 14 protein expression showed a declining trend with the increase of concentration,and the difference was statistically significant( all P < 0. 05). Conclusion 1,25( OH)_2D_3 can inhibit the invasion and migration of PANC-1cells through down-regulating the expression of MMP-2,9 and 14.
Objective To investigate the effect of 1,25-dihydroxyvitamin D_3[1,25(OH)_2D_3]on invasive and migratory abilities of human pancreatic cancer cell line PANC-1 and the changes in the expression of matrix metalloproteinases( MMP)-2,9 and 14. Methods The routinely cultured human pancreatic cancer cell line PANC-1 in vitro was treated with different concentrations of 1,25( OH)_2D_3( 0,25,50,75 and 100 μmol/L). After 24 hours,the abilities of migration and invasion in vitro in each group were detected by Transwell migration and invasion assays. After 48 hours,the protein expression of MMP-2,9 and 14 in each group was measured by Western blotting. Results After intervention for 24 h with different concentrations of 1,25( OH)_2D_3( 0,25,50,75 and 100 μmol/L),every field's invasive cells in each group were( 46. 20 ± 0. 84),( 30. 80 ± 2. 28),( 18. 80 ± 1. 64),( 13. 20 ± 2. 39) and( 5. 20 ± 1. 30) / HP; and the migratory cells in each group were( 66. 03 ± 1. 25),( 55. 82 ± 1. 65),( 41. 43 ± 1. 51),( 19. 86 ± 1. 45) and( 10. 23 ± 1. 95) /HP,respectively. The migratory and invasive cells were decreased with the increase of concentration,and the difference was statistically significant( all P < 0. 05). After intervention for 48 h with different concentrations of 1,25( OH)2D_3,MMP-2,9 and 14 protein expression showed a declining trend with the increase of concentration,and the difference was statistically significant( all P < 0. 05). Conclusion 1,25( OH)_2D_3 can inhibit the invasion and migration of PANC-1cells through down-regulating the expression of MMP-2,9 and 14.
引文
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[14]Ulasov I,Yi R,Guo D,et al.The emerging role of MMP-14 in brain tumorigenesis and future therapeutics[J].Biochim Biophys Acta,2014,1846(1):113-120.
[15]Hu B,Zhang K,Li S,et al.HIC1 attenuates invasion and metastasis by inhibiting the IL-6/STAT3 signalling pathway in human pancreatic cancer[J].Cancer Lett,2016,376(2):387-398.
[2]Ibrahim AM,Wang YH.Viro-immune therapy:a new strategy for treatment of pancreatic cancer[J].World J Gastroenterol,2016,22(2):748-763.
[3]Ryu JK.Recent advances in palliative chemotherapy for unresectable pancreatic cancer[J].Korean J Gastroenterol,2015,66(3):150-153.
[4]Chen S,Zhu J,Zuo S,et al.1,25(OH)2D3attenuates TGF-β1/β2-induced increased migration and invasion via inhibiting epithelial-mesenchymal transition in colon cancer cells[J].Biochem Biophys Res Commun,2015,468(1-2):130-135.
[5]Chiang KC,Kuo SF,Chen CH,et al.MART-10,the vitamin D analog,is a potent drug to inhibit anaplastic thyroid cancer cell metastatic potential[J].Cancer Lett,2015,369(1):76-85.
[6]Chiang KC,Chen SC,Yeh CN,et al.MART-10,a less calcemic vitamin D analog,is more potent than 1α,25-dihydroxyvitamin D3in inhibiting the metastatic potential of MCF-7 breast cancer cells in vitro[J].J Steroid Biochem Mol Biol,2014,139:54-60.
[7]Fink K,Boratyński J.The role of metalloproteinases in modification of extracellular matrix in invasive tumor growth,metastasis and angiogenesis[J].Postepy Hig Med Dosw(Online),2012,66(151):609-628.
[8]Torzilli PA,Bourne JW,Cigler T,et al.A new paradigm for mechanobiological mechanisms in tumor metastasis[J].Semin Cancer Biol,2012,22(5-6):385-395.
[9]Mittal R,Patel AP,Debs LH,et al.Intricate functions of matrix metalloproteinases in physiological and pathological conditions[J].J Cell Physiol,2016.[Epub ahead of print]
[10]Al-Alem L,Curry TE Jr.Ovarian cancer:involvement of the matrix metalloproteinases[J].Reproduction,2015,150(2):55-64.
[11]吴俊本,张洁,成丕光,等.胰腺癌组织中MMP-2、MMP-9的表达[J].中华胰腺病杂志,2012,12(1):30-32.
[12]Ren F,Tang R,Zhang X,et al.Overexpression of MMP family members functions as prognostic biomarker for breast cancer patients:a systematic review and meta-analysis[J].PLo S One,2015,10(8):e0135544.
[13]张振华,文娣娣,付欣,等.Survivin和MMP-14在胰腺癌中的表达及临床病理意义[J].现代生物医学进展,2015,15(16):3022-3027.
[14]Ulasov I,Yi R,Guo D,et al.The emerging role of MMP-14 in brain tumorigenesis and future therapeutics[J].Biochim Biophys Acta,2014,1846(1):113-120.
[15]Hu B,Zhang K,Li S,et al.HIC1 attenuates invasion and metastasis by inhibiting the IL-6/STAT3 signalling pathway in human pancreatic cancer[J].Cancer Lett,2016,376(2):387-398.