重楼皂苷Ⅰ通过下调CIP2A/PP2A/ERK信号通路抑制前列腺癌转移的机制研究
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摘要
目的:本文旨在探讨重楼皂苷Ⅰ(Polyphyllin I,PPI)对前列腺癌(prostate cancer,PC)细胞的增殖、转移及上皮-间质细胞转化(epithelial-mesenchymal transition,EMT)的抑制作用及具体分子机制。方法:通过实时荧光定量PCR法检测PC细胞(DU145和PC3)中EMT及转移相关基因的表达情况;通过MTT法及克隆形成实验检测PPI处理对细胞增殖能力的影响;通过Transwell以及划痕实验检测细胞的侵袭和迁移能力;通过Western blot检测转移相关蛋白的表达水平。结果:PPI处理后细胞的增殖能力显著下降,并具有显著的时间和剂量依赖性特点; PPI能够抑制PC细胞的侵袭、迁移并逆转EMT; PPI对细胞侵袭、迁移及EMT的抑制作用是通过CIP2A/PP2A/ERK信号通路来实现的。结论:在前列腺癌细胞中,PPI可能通过下调CIP2A/PP2A/ERK信号通路抑制肿瘤细胞的生长、转移及逆转EMT。因此,PPI可能成为前列腺癌治疗的新药物。
Objective This study was to investigate the inhibitory effect of Paris Polyphyllin Ⅰ(PPI) on proliferation,metastasis and epithelial-mesenchymal transition(EMT) of prostate cancer(PC) cells and its molecular mechanism.Methods The expression of EMT and metatastasis-related genes(DU145 and PC3) in PC cells were detected by real-time quantitative PCR(RT-q PCR).MTT and colony formation assay were used to detect proliferation,while transwell and wound healing assays were used to detect cell invasion and migration ability.Western blot was used to detect the expression levels of metastasis-related proteins.Results The cell proliferation was significantly decreased by PPI treatment in a dosage-and time-dependent manner.PPI could inhibit the invasion,migration and reversal of EMT in PC cells.The effects of PPI may be mediated by CIP2A/PP2A/ERK signaling pathway.Conclusion PPI may inhibit the growth,metastasis and reverse EMT of prostate cancer cells by down-regulating CIP2A/PP2A/ERK signaling pathway.Therefore,PPI may become a new drug for prostate cancer treatment.
Objective This study was to investigate the inhibitory effect of Paris Polyphyllin Ⅰ(PPI) on proliferation,metastasis and epithelial-mesenchymal transition(EMT) of prostate cancer(PC) cells and its molecular mechanism.Methods The expression of EMT and metatastasis-related genes(DU145 and PC3) in PC cells were detected by real-time quantitative PCR(RT-q PCR).MTT and colony formation assay were used to detect proliferation,while transwell and wound healing assays were used to detect cell invasion and migration ability.Western blot was used to detect the expression levels of metastasis-related proteins.Results The cell proliferation was significantly decreased by PPI treatment in a dosage-and time-dependent manner.PPI could inhibit the invasion,migration and reversal of EMT in PC cells.The effects of PPI may be mediated by CIP2A/PP2A/ERK signaling pathway.Conclusion PPI may inhibit the growth,metastasis and reverse EMT of prostate cancer cells by down-regulating CIP2A/PP2A/ERK signaling pathway.Therefore,PPI may become a new drug for prostate cancer treatment.
引文
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[15] Cai F,Zhang L,Xiao X,et al.Cucurbitacin B reverses multidrug resistance by targeting CIP2A to reactivate protein phosphatase 2A in MCF-7/adriamycin cells[J].Oncol Rep,2016,36(2):1 180-1 186.
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[18] Chen JC,Hsieh MJ,Chen CJ,et al. Polyphyllin G induce apoptosis and autophagy in human nasopharyngeal cancer cells by modulation of AKT and mitogen-activated protein kinase pathways in vitro and in vivo[J].Oncotarget,2016,7(43):70 276-70 289.
[19] He H,Zheng L,Sun YP,et al.Steroidal saponins from Paris polyphylla suppress adhesion,migration and invasion of human lung cancer A549 cells via down-regulating MMP-2 and MMP-9[J]. Asian Pac J Cancer Prev,2014,15(24):10 911-10 916.
[20] Yue G,Wei J,Qian X,et al. Synergistic anticancer effects of polyphyllin I and evodiamine on freshly-removed human gastric tumors[J].PloS one,2013,8(6):e65 164.
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[23] Chen CM,Lin CL,Chiou HL,et al.Loss of endothelial cell-specific molecule 1 promotes the tumorigenicity and metastasis of prostate cancer cells through regulation of the TIMP-1/MMP-9 expression[J]. Oncotarget,2017,8(8):13 886-13 897.
[24] Terracciano D,Mazzarella C,Di Carlo A,et al.Effects of the Erb B1/Erb B2 kinase inhibitor GW2974 on androgen-independent prostate cancer PC-3 cell line growth and NSE,chromogranin A and osteopontin content[J].Oncol Rep,2010,24(1):213-217.
[25] Feng T,Cao W,Shen W,et al. Arctigenin inhibits STAT3 and exhibits anticancer potential in human triple-negative breast cancer therapy[J].Oncotarget,2017,8(1):329-344.
[26] Liu Y,Dong Y,Zhang B,et al.Small compound 6-Oangeloylplenolin induces caspase-dependent apoptosis in human multiple myeloma cells[J]. Oncology letters,2013,6(2):556-558.
[27]张云飞,张特,马文静,等.神农架中草药博落回提取物血根碱的抗肿瘤作用研究进展[J].湖北医药学院学报,2018,37(5):476-483.
[28]袁晓宁,季娟丽,许佳欣,等.天然草药———癌症治疗中的潜在自噬诱导剂[J].现代肿瘤医学,2019,27(5):879-885.
[29] Zhang Q,Liu S,Parajuli KR,et al.Interleukin-17 promotes prostate cancer via MMP7-induced epithelial-to-mesenchymal transition[J]. Oncogene,2017,36(5):687-699.
[30] Cao Z,Koochekpour S,Strup SE,et al,Reversion of epithelial-mesenchymal transition by a novel agent DZ-50via IGF binding protein-3 in prostate cancer cells[J].Oncotarget,2017,8(45):78 507-78 519.
[31] Chang MC,Chen CA,Chen PJ,et al. Mesothelin enhances invasion of ovarian cancer by inducing MMP-7through MAPK/ERK and JNK pathways[J]. Biochem J,2012,442(2):293-302.
[32] Chen HN,Yuan K,Xie N,et al.PDLIM1 Stabilizes the E-Cadherin/beta-Catenin Complex to Prevent Epithelial-Mesenchymal Transition and Metastatic Potential of Colorectal Cancer Cells[J].Cancer Res,2016,76(5):1122-1 134.
[33] Andreolas C,Kalogeropoulou M,Voulgari A,et al.Fra-1 regulates vimentin during Ha-RAS-induced epithelial mesenchymal transition in human colon carcinoma cells[J].Int J Cancer,2008,122(8):1 745-1 756.
[34] Zhao J,Ou B,Han D,et al.Tumor-derived CXCL5 promotes human colorectal cancer metastasis through activation of the ERK/Elk-1/Snail and AKT/GSK3beta/beta-catenin pathways[J]. Mol Cancer,2017,16(1):70.
[35] Wen-Sheng W. ERK signaling pathway is involved in p15INK4b/p16INK4a expression and Hep G2 growth inhibition triggered by TPA and Saikosaponin a[J].Oncogene,2003,22(7):955-963.
[36] Rincon R,Cristobal I,Zazo S,et al.PP2A inhibition determines poor outcome and doxorubicin resistance in early breast cancer and its activation shows promising therapeutic effects[J].Oncotarget,2015,6(6):4 299-4 314.
[37] Tsukamoto S,Huang Y,Umeda D,et al.67-k Da laminin receptor-dependent protein phosphatase 2A(PP2A)activation elicits melanoma-specific antitumor activity overcoming drug resistance[J].J Biol Chem,2014,289(47):32 671-32 681.
[38] Wang J,Okkeri J,Pavic K,et al.Oncoprotein CIP2A is stabilized via interaction with tumor suppressor PP2A/B56[J].EMBO Rep,2017,18(3):437-450.
[2] Chen W,Zheng R,Baade PD,et al.Cancer statistics in China,2015[J].CA Cancer J Clin,2016,66(2):115-132.
[3] Siegel RL,Miller KD,Jemal A. Cancer statistics,2016[J].CA Cancer J Clin,2016,66(1):7-30.
[4]孙智婷,刘雪文,刘莹,等.癌蛋白CIP2A与多发性骨髓瘤患者预后不良相关性研究[J].湖北医药学院学报,2018,37(1):1-5,113.
[5] Junttila MR,Puustinen P,Niemela M,et al.CIP2A inhibits PP2A in human malignancies[J].Cell,2007,130(1):51-62.
[6] Khanna A,Bockelman C,Hemmes A,et al. MYC-dependent regulation and prognostic role of CIP2A in gastric cancer[J].J Natl Cancer Inst,2009,101(11):793-805.
[7] Teng HW,Yang SH,Lin JK,et al.CIP2A is a predictor of poor prognosis in colon cancer[J]. J Gastrointest Surg,2012,16(5):1 037-1 047.
[8] Vaarala MH,Vaisanen MR,Ristimaki A.CIP2A expression is increased in prostate cancer[J]. J Exp Clin Cancer Res,2010,29:136.
[9] Bockelman C,Lassus H,Hemmes A,et al. Prognostic role of CIP2A expression in serous ovarian cancer[J].Br J Cancer,2011,105(7):989-995.
[10] Tseng LM,Liu CY,Chang KC,et al.CIP2A is a target of bortezomib in human triple negative breast cancer cells[J].Breast Cancer Res,2012,14(2):R68.
[11] Liu Z,Ma L,Wen ZS,et al. Cancerous inhibitor of PP2A is targeted by natural compound celastrol for degradation in non-small-cell lung cancer[J].Carcinogenesis,2014,35(4):905-914.
[12] Come C,Laine A,Chanrion M,et al.CIP2A is associated with human breast cancer aggressivity[J]. Clin Cancer Res,2009,15(16):5 092-5 100.
[13] De P,Carlson JH,Leyland-Jones B,et al.Role of"oncogenic nexus"of CIP2A in breast oncogenesis:how does it work?[J]. Am J Cancer Res,2015,5(9):2872-2 891.
[14] Wu Y,Gu TT,Zheng PS.CIP2A cooperates with H-Ras to promote epithelial-mesenchymal transition in cervical-cancer progression[J]. Cancer letters,2015,356(2Pt B):646-655.
[15] Cai F,Zhang L,Xiao X,et al.Cucurbitacin B reverses multidrug resistance by targeting CIP2A to reactivate protein phosphatase 2A in MCF-7/adriamycin cells[J].Oncol Rep,2016,36(2):1 180-1 186.
[16] Xiao X,He Z,Cao W,et al.Oridonin inhibits gefitinibresistant lung cancer cells by suppressing EGFR/ERK/MMP-12 and CIP2A/Akt signaling pathways[J].Int J Oncol,2016,48(6):2 608-2 618.
[17] Li GB,Fu RQ,Shen HM,et al. Polyphyllin I induces mitophagic and apoptotic cell death in human breast cancer cells by increasing mitochondrial PINK1 levels[J].Oncotarget,2017,8(6):10 359-10 374.
[18] Chen JC,Hsieh MJ,Chen CJ,et al. Polyphyllin G induce apoptosis and autophagy in human nasopharyngeal cancer cells by modulation of AKT and mitogen-activated protein kinase pathways in vitro and in vivo[J].Oncotarget,2016,7(43):70 276-70 289.
[19] He H,Zheng L,Sun YP,et al.Steroidal saponins from Paris polyphylla suppress adhesion,migration and invasion of human lung cancer A549 cells via down-regulating MMP-2 and MMP-9[J]. Asian Pac J Cancer Prev,2014,15(24):10 911-10 916.
[20] Yue G,Wei J,Qian X,et al. Synergistic anticancer effects of polyphyllin I and evodiamine on freshly-removed human gastric tumors[J].PloS one,2013,8(6):e65 164.
[21] Shi YM,Yang L,Geng YD,et al.Polyphyllin I inducedapoptosis is enhanced by inhibition of autophagy in human hepatocellular carcinoma cells[J].Phytomedicine,2015,22(13):1 139-1 149.
[22] Livak KJ,Schmittgen TD.Analysis of relative gene expression data using real-time quantitative PCR and the2(-Delta Delta C(T))Method[J].Methods,2001,25(4):402-408.
[23] Chen CM,Lin CL,Chiou HL,et al.Loss of endothelial cell-specific molecule 1 promotes the tumorigenicity and metastasis of prostate cancer cells through regulation of the TIMP-1/MMP-9 expression[J]. Oncotarget,2017,8(8):13 886-13 897.
[24] Terracciano D,Mazzarella C,Di Carlo A,et al.Effects of the Erb B1/Erb B2 kinase inhibitor GW2974 on androgen-independent prostate cancer PC-3 cell line growth and NSE,chromogranin A and osteopontin content[J].Oncol Rep,2010,24(1):213-217.
[25] Feng T,Cao W,Shen W,et al. Arctigenin inhibits STAT3 and exhibits anticancer potential in human triple-negative breast cancer therapy[J].Oncotarget,2017,8(1):329-344.
[26] Liu Y,Dong Y,Zhang B,et al.Small compound 6-Oangeloylplenolin induces caspase-dependent apoptosis in human multiple myeloma cells[J]. Oncology letters,2013,6(2):556-558.
[27]张云飞,张特,马文静,等.神农架中草药博落回提取物血根碱的抗肿瘤作用研究进展[J].湖北医药学院学报,2018,37(5):476-483.
[28]袁晓宁,季娟丽,许佳欣,等.天然草药———癌症治疗中的潜在自噬诱导剂[J].现代肿瘤医学,2019,27(5):879-885.
[29] Zhang Q,Liu S,Parajuli KR,et al.Interleukin-17 promotes prostate cancer via MMP7-induced epithelial-to-mesenchymal transition[J]. Oncogene,2017,36(5):687-699.
[30] Cao Z,Koochekpour S,Strup SE,et al,Reversion of epithelial-mesenchymal transition by a novel agent DZ-50via IGF binding protein-3 in prostate cancer cells[J].Oncotarget,2017,8(45):78 507-78 519.
[31] Chang MC,Chen CA,Chen PJ,et al. Mesothelin enhances invasion of ovarian cancer by inducing MMP-7through MAPK/ERK and JNK pathways[J]. Biochem J,2012,442(2):293-302.
[32] Chen HN,Yuan K,Xie N,et al.PDLIM1 Stabilizes the E-Cadherin/beta-Catenin Complex to Prevent Epithelial-Mesenchymal Transition and Metastatic Potential of Colorectal Cancer Cells[J].Cancer Res,2016,76(5):1122-1 134.
[33] Andreolas C,Kalogeropoulou M,Voulgari A,et al.Fra-1 regulates vimentin during Ha-RAS-induced epithelial mesenchymal transition in human colon carcinoma cells[J].Int J Cancer,2008,122(8):1 745-1 756.
[34] Zhao J,Ou B,Han D,et al.Tumor-derived CXCL5 promotes human colorectal cancer metastasis through activation of the ERK/Elk-1/Snail and AKT/GSK3beta/beta-catenin pathways[J]. Mol Cancer,2017,16(1):70.
[35] Wen-Sheng W. ERK signaling pathway is involved in p15INK4b/p16INK4a expression and Hep G2 growth inhibition triggered by TPA and Saikosaponin a[J].Oncogene,2003,22(7):955-963.
[36] Rincon R,Cristobal I,Zazo S,et al.PP2A inhibition determines poor outcome and doxorubicin resistance in early breast cancer and its activation shows promising therapeutic effects[J].Oncotarget,2015,6(6):4 299-4 314.
[37] Tsukamoto S,Huang Y,Umeda D,et al.67-k Da laminin receptor-dependent protein phosphatase 2A(PP2A)activation elicits melanoma-specific antitumor activity overcoming drug resistance[J].J Biol Chem,2014,289(47):32 671-32 681.
[38] Wang J,Okkeri J,Pavic K,et al.Oncoprotein CIP2A is stabilized via interaction with tumor suppressor PP2A/B56[J].EMBO Rep,2017,18(3):437-450.