雷公藤红素对人胰腺癌细胞PANC-1增殖、侵袭和迁移的抑制作用
|
摘要
背景与目的:雷公藤红素(celastrol)是雷公藤的主要活性成分,现代研究发现其具有广泛的抗癌活性,对胰腺癌细胞凋亡有一定的促进作用。该研究旨在探讨celastrol对胰腺癌细胞PANC-1增殖、侵袭和迁移的作用。方法:将PANC-1细胞分为PANC-1组、celastrol(5μmol/L)组、celastrol(10μmol/L)组和celastrol(20μmol/L)组,分别用0、5、10和20μmol/L的celastrol处理PANC-1细胞,细胞计数试剂盒(cell counting kit-8,CCK-8)检测细胞增殖倍数,Transwell检测各组PANC-1细胞的侵袭能力,划痕实验检测各组细胞迁移能力。蛋白质印迹法(Western blot)检测Ki-67、血管内皮生长因子(vascular endothelial growth factor,VEGF)和基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)的蛋白表达水平。结果:与PANC-1组比较,celastrol(5μmol/L)组、celastrol(10μmol/L)组和celastrol(20μmol/L)组细胞增殖倍数明显降低,细胞增殖标记蛋白Ki-67的蛋白表达水平与PANC-1组比较也明显降低;同时,celastrol(5μmol/L)组、celastrol(10μmol/L)组和celastrol(20μmol/L)组侵袭细胞数与PANC-1组相比明显减少;此外,与PANC-1组比较,celastrol(5μmol/L)组、celastrol(10μmol/L)组和celastrol(20μmol/L)组划痕闭合率显著降低,VEGF和MMP-9的蛋白表达水平也显著低于PANC-1组。结论:Celastrol能抑制人胰腺癌细胞PANC-1增殖、侵袭及迁移。
Background and purpose: Celastrol is the primary active component of Tripterygium wilfordii. The present studies indicate that celastrol displays extensive antitumor activity and promotes apoptosis in pancreatic cancer. This study aimed to investigate the effect of celastrol on cell proliferation, invasion and migration of human pancreatic cancer cell line PANC-1. Methods: PANC-1 cells were divided into PANC-1 group, celastrol (5 μmol/L) group, celastrol (10 μmol/L) group and celastrol (20 μmol/L) group, and the cells were treated with different levels of celastrol (5, 10 and 20 μmol/L). Cell counting kit-8 (CCK-8) was used to evaluate cell proliferation. The invasion ability was determined using Transwell assay, and cell migration ability was determined by wound healing assay. The expressions of proteins, including Ki-67, vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) were measured by Western blot. Results: Compared with PANC-1 group, the cell proliferation was decreased in celastrol (5 μmol/L) group, celastrol (10 μmol/L) group and celastrol (20 μmol/L) group, and the protein level of Ki-67 was decreased significantly in celastrol-treated groups compared with PANC-1 group. Meanwhile, cell invasion was decreased in celastrol (5 μmol/L) group, celastrol (10 μmol/L) group and celastrol (20 μmol/L) group compared with PANC-1 group. In addition, compared with PANC-1 group, the wound closure rates in celastrol (5 μmol/L) group, celastrol (10 μmol/L) group and celastrol (20 μmol/L) group were decreased notably, and the protein levels of VEGF and MMP-9 were decreased compared with PANC-1 group. Conclusion: Celastrol can inhibit cell proliferation, invasion and migration of human pancreatic cancer cell line PANC-1.
Background and purpose: Celastrol is the primary active component of Tripterygium wilfordii. The present studies indicate that celastrol displays extensive antitumor activity and promotes apoptosis in pancreatic cancer. This study aimed to investigate the effect of celastrol on cell proliferation, invasion and migration of human pancreatic cancer cell line PANC-1. Methods: PANC-1 cells were divided into PANC-1 group, celastrol (5 μmol/L) group, celastrol (10 μmol/L) group and celastrol (20 μmol/L) group, and the cells were treated with different levels of celastrol (5, 10 and 20 μmol/L). Cell counting kit-8 (CCK-8) was used to evaluate cell proliferation. The invasion ability was determined using Transwell assay, and cell migration ability was determined by wound healing assay. The expressions of proteins, including Ki-67, vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) were measured by Western blot. Results: Compared with PANC-1 group, the cell proliferation was decreased in celastrol (5 μmol/L) group, celastrol (10 μmol/L) group and celastrol (20 μmol/L) group, and the protein level of Ki-67 was decreased significantly in celastrol-treated groups compared with PANC-1 group. Meanwhile, cell invasion was decreased in celastrol (5 μmol/L) group, celastrol (10 μmol/L) group and celastrol (20 μmol/L) group compared with PANC-1 group. In addition, compared with PANC-1 group, the wound closure rates in celastrol (5 μmol/L) group, celastrol (10 μmol/L) group and celastrol (20 μmol/L) group were decreased notably, and the protein levels of VEGF and MMP-9 were decreased compared with PANC-1 group. Conclusion: Celastrol can inhibit cell proliferation, invasion and migration of human pancreatic cancer cell line PANC-1.
引文
[1]CAO L,CHEN X,XIAO X,et al.Resveratrol inhibits hyperglycemia-driven ROS-induced invasion and migration of pancreatic cancer cells via suppression of the ERK and p38MAPK signaling pathways[J].Int J Oncol,2016,49(2):735-743.
[2]PEKALA K R,MA X,KROPP P A,et al.Loss of HNF6expression correlates with human pancreatic cancer progression[J].Lab Invest,2014,94(5):517-527.
[3]CHEN W,ZHENG R,BAADE P D,et al.Cancer statistics in China,2015[J].Ca Cancer J Clin,2016,66(2):115-132.
[4]ZHENG W,LU S,CAI H,et al.Deguelin inhibits proliferation and migration of human pancreatic cancer cells in vitro targeting hedgehog pathway[J].Oncol Lett,2016,12(4):2761-2765.
[5]CHEN L,QU C,CHEN H,et al.Chinese herbal medicine suppresses invasion-promoting capacity of cancer-associated fibroblasts in pancreatic cancer[J].PLoS One,2014,9(4):e96177.
[6]GUO J,HUANG X,WANG H,et al.Celastrol induces autophagy by targeting AR/miR-101 in prostate cancer cells[J].PLoS One,2015,10(10):e0140745.
[7]XU J,WU C L.Anti-metastasis of celastrol on esophageal cancer cells and its mechanism[J].Sheng LI Xue Bao,2015,67(3):341-347.
[8]SHA M,YE J,LUAN Z,et al.Celastrol induces cell cycle arrest by MicroRNA-21-mTOR-mediated inhibition p27 protein degradation in gastric cancer[J].Cancer Cell Inter,2015,15:101.
[9]ZHAO X,GAO S,REN H,et al.Inhibition of autophagy strengthens celastrol-induced apoptosis in human pancreatic cancer in vitro and in vivo models[J].Curr Mol Med,2014,14(4):555-563.
[10]XU Q,ZONG L,CHEN X,et al.Resveratrol in the treatment of pancreatic cancer[J].Ann N Y Acad Sci,2015,1348(1):10-19.
[11]LAHIANI M H,EASSA S,PARNELL C,et al.Carbon nanotubes as carriers of Panax ginseng metabolites and enhancers of ginsenosides Rb1 and Rg1 anti-cancer activity[J].Nanotechnology,2017,28(1):015101.
[12]SHRIVASTAVA S,JEENGAR M K,Reddy V S,et al.Anticancer effect of celastrol on human triple negative breast cancer:possible involvement of oxidative stress,mitochondrial dysfunction,apoptosis and PI3K/Akt pathways[J].Exp Mol Pathol,2015,98(3):313-327.
[13]PANG X,YI Z,ZHANG J,et al.Celastrol suppresses angiogenesis-mediated tumor growth through inhibition of AKT/mammalian target of rapamycin pathway[J].Cancer Res,2010,70(5):1951-1959.
[14]LEE H W,JANG K S,CHOI H J,et al.Celastrol inhibits gastric cancer growth by induction of apoptosis and autophagy[J].Bmb Reports,2014,47(12):697-702.
[15]XU J,WU C L,HUANG J.Effect of celastrol in inhibiting metastasis of lung cancer cells by influencing Akt signaling pathway and expressing integrins[J].Zhongguo Zhong Yao Za Zhi,2015,40(6):1129-1133.
[16]CHAKRAVARTHY R,CLEMENS M J,PIRIANOV G,et al.Role of the eIF4E binding protein 4E-BP1 in regulation of the sensitivity of human pancreatic cancer cells to TRAIL and celastrol-induced apoptosis[J].Biol Cell,2013,105(9):414-429.
[17]RAZIDLO G L,MAGNINE C,SLETTEN A C,et al.Targeting pancreatic cancer metastasis by inhibition of vav1,a driver of tumor cell invasion[J].Cancer Res,2015,75(14):2907-2915.
[18]YU X,WANG Q,ZHOU X,et al.Celastrol negatively regulates cell invasion and migration ability of human osteosarcoma via downregulation of the PI3K/Akt/NF-κB signaling pathway in vitro[J].Oncol Lett,2016,12(5):3423-3428.
[19]WANG Z,ZHAI Z,DU X.Celastrol inhibits migration and invasion through blocking the NF-κB pathway in ovarian cancer cells[J].Exp Ther Med,2017,14(1):819-824.
[20]KANG H,JANG SW,KO J.Celastrol inhibits breast cancer cell invasion via suppression of NF-κB-mediated matrix metalloproteinase-9 expression[J].Cell Physiol Biochem,2011,28(2):175-184.
[21]AKTER H,PARK M,KWON O S,et al.Activation of matrix metalloproteinase-9(MMP-9) by neurotensin promotes cell invasion and migration through ERK pathway in gastric cancer[J].Tumour Biol,2015,36(8):6053-6062.
[22]LI Y,ZHAI Z,LIU D,et al.CD105 promotes hepatocarcinoma cell invasion and metastasis through VEGF[J].Tumour Biol,2015,36(2):737-745.
[2]PEKALA K R,MA X,KROPP P A,et al.Loss of HNF6expression correlates with human pancreatic cancer progression[J].Lab Invest,2014,94(5):517-527.
[3]CHEN W,ZHENG R,BAADE P D,et al.Cancer statistics in China,2015[J].Ca Cancer J Clin,2016,66(2):115-132.
[4]ZHENG W,LU S,CAI H,et al.Deguelin inhibits proliferation and migration of human pancreatic cancer cells in vitro targeting hedgehog pathway[J].Oncol Lett,2016,12(4):2761-2765.
[5]CHEN L,QU C,CHEN H,et al.Chinese herbal medicine suppresses invasion-promoting capacity of cancer-associated fibroblasts in pancreatic cancer[J].PLoS One,2014,9(4):e96177.
[6]GUO J,HUANG X,WANG H,et al.Celastrol induces autophagy by targeting AR/miR-101 in prostate cancer cells[J].PLoS One,2015,10(10):e0140745.
[7]XU J,WU C L.Anti-metastasis of celastrol on esophageal cancer cells and its mechanism[J].Sheng LI Xue Bao,2015,67(3):341-347.
[8]SHA M,YE J,LUAN Z,et al.Celastrol induces cell cycle arrest by MicroRNA-21-mTOR-mediated inhibition p27 protein degradation in gastric cancer[J].Cancer Cell Inter,2015,15:101.
[9]ZHAO X,GAO S,REN H,et al.Inhibition of autophagy strengthens celastrol-induced apoptosis in human pancreatic cancer in vitro and in vivo models[J].Curr Mol Med,2014,14(4):555-563.
[10]XU Q,ZONG L,CHEN X,et al.Resveratrol in the treatment of pancreatic cancer[J].Ann N Y Acad Sci,2015,1348(1):10-19.
[11]LAHIANI M H,EASSA S,PARNELL C,et al.Carbon nanotubes as carriers of Panax ginseng metabolites and enhancers of ginsenosides Rb1 and Rg1 anti-cancer activity[J].Nanotechnology,2017,28(1):015101.
[12]SHRIVASTAVA S,JEENGAR M K,Reddy V S,et al.Anticancer effect of celastrol on human triple negative breast cancer:possible involvement of oxidative stress,mitochondrial dysfunction,apoptosis and PI3K/Akt pathways[J].Exp Mol Pathol,2015,98(3):313-327.
[13]PANG X,YI Z,ZHANG J,et al.Celastrol suppresses angiogenesis-mediated tumor growth through inhibition of AKT/mammalian target of rapamycin pathway[J].Cancer Res,2010,70(5):1951-1959.
[14]LEE H W,JANG K S,CHOI H J,et al.Celastrol inhibits gastric cancer growth by induction of apoptosis and autophagy[J].Bmb Reports,2014,47(12):697-702.
[15]XU J,WU C L,HUANG J.Effect of celastrol in inhibiting metastasis of lung cancer cells by influencing Akt signaling pathway and expressing integrins[J].Zhongguo Zhong Yao Za Zhi,2015,40(6):1129-1133.
[16]CHAKRAVARTHY R,CLEMENS M J,PIRIANOV G,et al.Role of the eIF4E binding protein 4E-BP1 in regulation of the sensitivity of human pancreatic cancer cells to TRAIL and celastrol-induced apoptosis[J].Biol Cell,2013,105(9):414-429.
[17]RAZIDLO G L,MAGNINE C,SLETTEN A C,et al.Targeting pancreatic cancer metastasis by inhibition of vav1,a driver of tumor cell invasion[J].Cancer Res,2015,75(14):2907-2915.
[18]YU X,WANG Q,ZHOU X,et al.Celastrol negatively regulates cell invasion and migration ability of human osteosarcoma via downregulation of the PI3K/Akt/NF-κB signaling pathway in vitro[J].Oncol Lett,2016,12(5):3423-3428.
[19]WANG Z,ZHAI Z,DU X.Celastrol inhibits migration and invasion through blocking the NF-κB pathway in ovarian cancer cells[J].Exp Ther Med,2017,14(1):819-824.
[20]KANG H,JANG SW,KO J.Celastrol inhibits breast cancer cell invasion via suppression of NF-κB-mediated matrix metalloproteinase-9 expression[J].Cell Physiol Biochem,2011,28(2):175-184.
[21]AKTER H,PARK M,KWON O S,et al.Activation of matrix metalloproteinase-9(MMP-9) by neurotensin promotes cell invasion and migration through ERK pathway in gastric cancer[J].Tumour Biol,2015,36(8):6053-6062.
[22]LI Y,ZHAI Z,LIU D,et al.CD105 promotes hepatocarcinoma cell invasion and metastasis through VEGF[J].Tumour Biol,2015,36(2):737-745.