槲皮素通过抑制β-catenin入核抑制人宫颈癌细胞增殖、迁移的实验研究
|
摘要
[目的]探讨槲皮素通过Wnt/β-catenin信号通路对人宫颈癌上皮细胞增殖及迁移的影响及其可能的作用机制,为抑制宫颈癌进展的临床治疗提供新的靶点。[方法]人宫颈癌上皮细胞系HeLa细胞为研究对象,给予不同浓度槲皮素处理24 h后,通过CCK-8实验检测槲皮素对HeLa细胞增殖能力的影响。采用Transwell及细胞划痕实验检测槲皮素对HeLa细胞垂直及水平运动能力的影响。免疫荧光法检测β-catenin表达及TopFlash荧光素酶报告系统检测槲皮素对HeLa细胞Wnt/β-catenin信号通路的抑制作用。[结果] CCK-8检测表明,槲皮素处理组HeLa细胞增殖率明显低于对照组(P<0.01),且抑制作用具有浓度依赖性。槲皮素处理组HeLa细胞垂直及水平运动能力均较对照细胞降低,差异有统计学意义(P<0.01)。对照组HeLa细胞中β-catenin蛋白密集分布于胞核和胞质中,而槲皮素处理组中β-catenin蛋白主要分布于细胞质中。TopFlash荧光素酶报告系统表明槲皮素处理组HeLa细胞Wnt/β-catenin信号通路受到抑制。[结论]槲皮素可能通过抑制Wnt/β-catenin信号通路活化抑制宫颈癌细胞的增殖和迁移。
[Objective] To investigate the effects of quercetinon proliferation and migration of human cervical cancer(CC) cells by Wnt/β-catenin signaling pathway and its mechanism and to provide a new gene target in the treatment of CC. [ Methods] The research objects were the human CC HeLa cells. After treatment with different concentrations ofquercetin for 24 hours, the proliferation of HeLa cells was detected by CCK-8 test, the horizontal and vertical migration were detected by Transwell and wound scraching assays, the activation of Wnt/β-catenin signaling pathway was detected by β-catenin immunofluorescence assay and TOP-flash luciferase reporter system.[Results] CCK-8 test indicated that the cell proliferation rate of the quercetin group was significantly lower than the control group(P<0.01),and the inhibition of quercetinon proliferation was dose-dependent. The horizontal and vertical migration of Hela cells in the quercetin group was significantly decreased compared to the control group(P<0.01). The β-catenin immunofluorescence was distributed in nuclear and cytoplasm in the control cells, while mainly in the cytoplasm in the quercetin-treated cells. Top Flashluciferase reporter system indicated the Wnt/β-catenin signaling pathway was inhibited in the quercetin-treated cells. [Conclusion] Quercetin may inhibit proliferation and migration of human cervical cancer cells via inhibiting Wnt/β-catenin signaling pathway.
[Objective] To investigate the effects of quercetinon proliferation and migration of human cervical cancer(CC) cells by Wnt/β-catenin signaling pathway and its mechanism and to provide a new gene target in the treatment of CC. [ Methods] The research objects were the human CC HeLa cells. After treatment with different concentrations ofquercetin for 24 hours, the proliferation of HeLa cells was detected by CCK-8 test, the horizontal and vertical migration were detected by Transwell and wound scraching assays, the activation of Wnt/β-catenin signaling pathway was detected by β-catenin immunofluorescence assay and TOP-flash luciferase reporter system.[Results] CCK-8 test indicated that the cell proliferation rate of the quercetin group was significantly lower than the control group(P<0.01),and the inhibition of quercetinon proliferation was dose-dependent. The horizontal and vertical migration of Hela cells in the quercetin group was significantly decreased compared to the control group(P<0.01). The β-catenin immunofluorescence was distributed in nuclear and cytoplasm in the control cells, while mainly in the cytoplasm in the quercetin-treated cells. Top Flashluciferase reporter system indicated the Wnt/β-catenin signaling pathway was inhibited in the quercetin-treated cells. [Conclusion] Quercetin may inhibit proliferation and migration of human cervical cancer cells via inhibiting Wnt/β-catenin signaling pathway.
引文
[1]Torre LA,Bray F,Siegel RL,et al.Global cancer statistics,2012[J].CA Cancer J Clin,2015,65(2):87-108.
[2]Andreucci M,Faga T,Pisani A,et al.Quercetin protects against radiocontrast medium toxicity in human renal proximal tubular cells[J].J Cell Physiol,2018,233(5):4116-4125.
[3]Sharma A,Kashyap D,Sak K,et al.Therapeutic charm of quercetin and its derivatives:a review of research and patents[J].Pharm Pat Anal,2018,7(1):15-32.
[4]Wilsher NE,Arroo RR,Matsoukas MT,et al.Cytochrome P450CYP1 metabolism of hydroxylated flavones and flavonols:Selective bioactivation of luteolin in breast cancer cells[J].Food Chem Toxicol,2017,110:383-394.
[5]Wang P,Henning SM,Magyar CE,et al.Green tea and quercetin sensitize PC-3 xenograft prostate tumors to docetaxel chemotherapy[J].J Exp Clin Cancer Res,2016,35(1):1-11.
[6]Lee WJ,Chen YR and Tseng TH.Quercetin induces FasL-related apoptosis,in part,through promotion of histone H3 acetylation in human leukemia HL-60 cells[J].Oncol Rep,2011,25(2):583-591.
[7]Li H,Jiao S,Li X,et al.Therapeutic effects of antibiotic drug tigecycline against cervical squamous cell carcinoma by inhibiting Wnt/beta-catenin signaling[J].Biochem Biophys Res Commun 2015,467(1):14-20.
[8]Qi L,Sun B,Liu Z,et al.Dickkopf-1 inhibits epithelialmesenchymal transition of colon cancer cells and contributes to colon cancer suppression[J].Cancer Sci,2012,103(4):828-835.
[9]Granato M,Rizzello C,Gilardini Montani MS,et al.Quercetin induces apoptosis and autophagy in primary effusion lymphoma cells by inhibiting PI3K/AKT/mTOR and STAT3 signaling pathways[J].JNutr Biochem,2017(41):124-136.
[10]Kim H,Seo EM,Sharma AR,et al.Regulation of Wnt signaling activity for growth suppression induced by quercetin in 4T1 murine mammary cancer cells[J].Int J Oncol,2013,43(4):1319-1325.
[11]Ahmed HH,Aglan HA,Zaazaa AM,et al.Quercetin confers tumoricidal activity through multipathway mechanisms in a N-methylnitrosourea rat model of colon cancer[J].Asian Pac J Cancer Prev,2016,17(11):4991-4998.
[12]Bishayee K,Ghosh S,Mukherjee A,et al.Quercetin induces cytochrome-c release and ROS accumulation to promote apoptosis and arrest the cell cycle in G2/M,in cervical carcinoma:signal cascade and drug-DNA interaction[J].Cell Prolif,2013,46(2):153-163.
[13]陈娇,张蔚,敖良飞,等.槲皮素对宫颈癌HeLa细胞STAT3的表达及其信号通路的影响[J].中华临床医师杂志(电子版),2011,5(19):5656-5661.
[14]程静,张蔚,吕琼莹,等.槲皮素和乙酰肝素酶与宫颈癌细胞侵袭与迁移机制的研究[J].实妇产科杂志,2014,30(3):212-214.
[15]陈眣冰,崔元璐.载槲皮素-粉防己碱纳米凝胶的制备、表征与体外评价[J].天津中医,2017,34(11):770-774.
[16]Bahrami A,Hasanzadeh M,ShahidSales S,et al.Clinical significance and prognosis value of Wnt signaling pathway in cervical cancer[J].J Cell Biochem 2017,118:3028-3033.
[17]Wei H,Wang N,Zhang Y,et al.Wnt-11 overexpression promoting the invasion of cervical cancer cells[J].Tumour Biol,2016,37(9):11789-11798.
[2]Andreucci M,Faga T,Pisani A,et al.Quercetin protects against radiocontrast medium toxicity in human renal proximal tubular cells[J].J Cell Physiol,2018,233(5):4116-4125.
[3]Sharma A,Kashyap D,Sak K,et al.Therapeutic charm of quercetin and its derivatives:a review of research and patents[J].Pharm Pat Anal,2018,7(1):15-32.
[4]Wilsher NE,Arroo RR,Matsoukas MT,et al.Cytochrome P450CYP1 metabolism of hydroxylated flavones and flavonols:Selective bioactivation of luteolin in breast cancer cells[J].Food Chem Toxicol,2017,110:383-394.
[5]Wang P,Henning SM,Magyar CE,et al.Green tea and quercetin sensitize PC-3 xenograft prostate tumors to docetaxel chemotherapy[J].J Exp Clin Cancer Res,2016,35(1):1-11.
[6]Lee WJ,Chen YR and Tseng TH.Quercetin induces FasL-related apoptosis,in part,through promotion of histone H3 acetylation in human leukemia HL-60 cells[J].Oncol Rep,2011,25(2):583-591.
[7]Li H,Jiao S,Li X,et al.Therapeutic effects of antibiotic drug tigecycline against cervical squamous cell carcinoma by inhibiting Wnt/beta-catenin signaling[J].Biochem Biophys Res Commun 2015,467(1):14-20.
[8]Qi L,Sun B,Liu Z,et al.Dickkopf-1 inhibits epithelialmesenchymal transition of colon cancer cells and contributes to colon cancer suppression[J].Cancer Sci,2012,103(4):828-835.
[9]Granato M,Rizzello C,Gilardini Montani MS,et al.Quercetin induces apoptosis and autophagy in primary effusion lymphoma cells by inhibiting PI3K/AKT/mTOR and STAT3 signaling pathways[J].JNutr Biochem,2017(41):124-136.
[10]Kim H,Seo EM,Sharma AR,et al.Regulation of Wnt signaling activity for growth suppression induced by quercetin in 4T1 murine mammary cancer cells[J].Int J Oncol,2013,43(4):1319-1325.
[11]Ahmed HH,Aglan HA,Zaazaa AM,et al.Quercetin confers tumoricidal activity through multipathway mechanisms in a N-methylnitrosourea rat model of colon cancer[J].Asian Pac J Cancer Prev,2016,17(11):4991-4998.
[12]Bishayee K,Ghosh S,Mukherjee A,et al.Quercetin induces cytochrome-c release and ROS accumulation to promote apoptosis and arrest the cell cycle in G2/M,in cervical carcinoma:signal cascade and drug-DNA interaction[J].Cell Prolif,2013,46(2):153-163.
[13]陈娇,张蔚,敖良飞,等.槲皮素对宫颈癌HeLa细胞STAT3的表达及其信号通路的影响[J].中华临床医师杂志(电子版),2011,5(19):5656-5661.
[14]程静,张蔚,吕琼莹,等.槲皮素和乙酰肝素酶与宫颈癌细胞侵袭与迁移机制的研究[J].实妇产科杂志,2014,30(3):212-214.
[15]陈眣冰,崔元璐.载槲皮素-粉防己碱纳米凝胶的制备、表征与体外评价[J].天津中医,2017,34(11):770-774.
[16]Bahrami A,Hasanzadeh M,ShahidSales S,et al.Clinical significance and prognosis value of Wnt signaling pathway in cervical cancer[J].J Cell Biochem 2017,118:3028-3033.
[17]Wei H,Wang N,Zhang Y,et al.Wnt-11 overexpression promoting the invasion of cervical cancer cells[J].Tumour Biol,2016,37(9):11789-11798.