[1]李东泽,吴良邦△.槲皮素通过小分子核糖核酸196b调节磷脂酰肌醇激酶/蛋白激酶B/雷帕霉素靶蛋白通路从而抑制骨肉瘤细胞增殖研究[J].中国中医骨伤科杂志,2023,31(06):8-14.[doi:10.20085/j.cnki.issn1005-0205.230602]
　LI Dongze,WU Liangbang.Quercetin Inhibits the Proliferation of Osteosarcoma Cells by Regulating miR196b-PI3K/AKT/mTOR Pathway[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2023,31(06):8-14.[doi:10.20085/j.cnki.issn1005-0205.230602]
点击复制

槲皮素通过小分子核糖核酸196b调节磷脂酰肌醇激酶/蛋白激酶B/雷帕霉素靶蛋白通路从而抑制骨肉瘤细胞增殖研究()
分享到：

《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第31卷
期数:
2023年06期
页码:
8-14
栏目:
实验研究
出版日期:
2023-06-15

文章信息/Info

Title:
Quercetin Inhibits the Proliferation of Osteosarcoma Cells by Regulating miR196b-PI3K/AKT/mTOR Pathway
文章编号:
1005-0205(2023)06-0008-07
作者:
李东泽1; 吴良邦1△
1解放军联勤保障部队第九〇三医院骨科(杭州,310009)
Author(s):
LI Dongze1; WU Liangbang1△
1Orthopedics Department of the 903 Hospital of the PLA Joint Service Support Force,Hangzhou 310009,China.
关键词:
槲皮素;  骨肉瘤细胞;  小分子核糖核酸196b;  磷脂酰肌醇激酶/蛋白激酶B/雷帕霉素靶蛋白信号通路
Keywords:
quercetin;  osteosarcoma cells;  miR-196b;  PI3K/AKT/mTOR signaling pathway
分类号:
R738.1
DOI:
10.20085/j.cnki.issn1005-0205.230602
文献标志码:
A
摘要:
目的:探讨槲皮素抗骨肉瘤细胞增殖的可能机制。方法:体外培养骨肉瘤细胞系MG-63、143B和U2OS,用25,50和100 μmol/L的槲皮素干预后,通过实时荧光定量聚合酶链式反应(RT-PCR)检测小分子核糖核酸196b(miR-196b)的水平。随后利用miR-196b抑制剂,联用100 μmol/L槲皮素后通过CCK8检测细胞的存活率; 通过划痕和Transwell实验检测细胞的迁移和侵袭; 通过蛋白质印迹法(Western Blot)检测细胞中磷脂酰肌醇激酶/蛋白激酶B/雷帕霉素靶蛋白(PI3K/AKT/mTOR)信号通路的表达。结果:槲皮素上调了三种骨肉瘤细胞中miR-196b的水平,且呈现剂量依赖性。在应用miR-196b抑制剂后,miR-196b的表达水平被显著抑制。槲皮素能够显著抑制骨肉瘤细胞的生长,但在联用miR-196b抑制剂后,槲皮素抑制骨肉瘤细胞增殖的作用被减弱。划痕和Transwell实验结果显示,槲皮素能够显著抑制骨肉瘤细胞的迁移和侵袭,但在联用miR-196b抑制剂后,槲皮素抑制骨肉瘤细胞迁移和侵袭的作用被减弱。此外,槲皮素显著抑制了骨肉瘤细胞中p-mTOR、p-AKT和p-PI3K的表达,对mTOR、AKT和PI3K的表达则无明显影响。联用miR-196b抑制剂后,p-mTOR、p-AKT和p-PI3K的表达显著上升,与单用槲皮素组相比差异有统计学意义(P<0.05)。结论:槲皮素通过上调miR-196b调节PI3K/AKT/mTOR信号通路,从而抑制骨肉瘤细胞的增殖、迁移和侵袭。
Abstract:
Objective:To evaluate the anti-proliferation effect of quercetin on osteosarcoma cells and the related mechanism.Methods:Osteosarcoma cell lines MG-63,143B and U2OS were cultured in vitro.After the treatment with 25,50 and 100 μmol/L quercetin,the level of miR-196b was detected by real-time fluorescence quantitative polymerase chain reaction(RT-PCR).Then,the survival rate of cells was detected by CCK8 after using miR-196b inhibitor combined with 100 μmol/L quercetin.Cell migration and invasion were detected by wound healing and Transwell assay.The expressions of phosphatidylinositol kinase/protein kinase B/rapamycin target protein(PI3K/AKT/mTOR)signal pathway were detected by Western Blot.Results:Quercetin upregulated the level of miR-196b in three osteosarcoma cells in a dose-dependent manner.After using miR-196b inhibitor,the expression level of miR-196b was significantly inhibited.Quercetin can significantly inhibit the growth of osteosarcoma cells,but the inhibitory effect was weakened when combined with miR-196b inhibitor.The results of wound healing and Transwell assay showed that quercetin could significantly inhibit the migration and invasion of osteosarcoma cells.However,when combined with miR-196b inhibitor,the effect of quercetin on the migration and invasion of osteosarcoma cells was weakened.In addition,quercetin significantly inhibited the expressions of p-mTOR,p-AKT and p-PI3K in osteosarcoma cells,but had no significant effect on the expressions of mTOR,AKT and PI3K.The expression of p-mTOR,p-AKT and p-PI3K increased significantly after the combination of miR-196b inhibitor,which was significantly different from that of quercetin alone.Conclusion:Quercetin regulates PI3K/AKT/mTOR signaling pathway by upregulating miR-196b,thereby inhibiting the proliferation,migration and invasion of osteosarcoma cells.

参考文献/References:

[1] WHELAN J,MCTIERNAN A,COOPER N,et al.Incidence and survival of malignant bone sarcomas in England 1979-2007[J].Int J Cancer,2012,131(4):E508-E517.
[2] 中国临床肿瘤学会肉瘤专家委员会,中国抗癌协会肿瘤临床化疗专业委员会.中国蒽环类药物治疗骨肉瘤专家共识[J].癌症,2022,41(8):368-384.
[3] SHENG G,GAO Y,YANG Y,et al.Osteosarcoma and metastasis[J].Front Oncol,2021,11:780264.
[4] KIM V N,NAM J W.Genomics of microRNA[J].Trends Genet,2006,22(3):165-173.
[5] 袁长深,容伟明,卢智贤,等.基于生物信息学构建骨肉瘤miRNA-mRNA的调控网络[J].中国组织工程研究,2021,25(17):2740-2746.
[6] CAI H,ZHAO H,TANG J,et al.Serum miR-195 is a diagnostic and prognostic marker for osteosarcoma[J].J Surg Res,2015,194(2):505-510.
[7] 刘念,张红,薛中柱.miRNA-199a-5p对骨肉瘤细胞增殖及自噬的影响[J].肿瘤防治研究,2017,44(3):197-201.
[8] ISLAM M S,QUISPE C,HOSSAIN R,et al.Neuropharmacological effects of quercetin:a literature-based review[J].Front Pharmacol,2021,12:665031.
[9] ASGHARIAN P,TAZEKAND A P,HOSSEINI K,et al.Potential mechanisms of quercetin in cancer prevention:focus on cellular and molecular targets[J].Cancer Cell Int,2022,22(1):257.
[10] 申昌梅,廖振蓉,余瑛.基于PI3K/Akt信号通路探讨槲皮素增强顺铂对人宫颈癌顺铂耐药细胞HeLa/DDP的作用[J].中成药,2022,44(11):3667-3671.
[11] KHAN F,NIAZ K,MAQBOOL F,et al.Molecular targets underlying the anticancer effects of quercetin:an update[J].Nutrients,2016,8(9):529.
[12] KARSY M,ARSLAN E,MOY F.Current progress on understanding microRNAs in glioblastoma multiforme[J].Genes Cancer,2012,3(1):3-15.
[13] ZHANG C,YAO C,LI H,et al.Combined elevation of microRNA-196a and microRNA-196b in sera predicts unfavorable prognosis in patients with osteosarcomas[J].Int J Mol Sci,2014,15(4):6544-6555.
[14] 李欣,林明哲,赵久达.槲皮素对胃癌相关p53/AMPK/mTOR信号通路的影响[J].天津医药,2021,49(11):1143-1147.
[15] SHEN X,SI Y,WANG Z,et al.Quercetin inhibits the growth of human gastric cancer stem cells by inducing mitochondrial-dependent apoptosis through the inhibition of PI3K/Akt signaling[J].Int J Mol Med,2016,38(2):619-626.
[16] NWAEBURU C C,BAUER N,ZHAO Z,et al.Up-regulation of microRNA let-7c by quercetin inhibits pancreatic cancer progression by activation of Numbl[J].Oncotarget,2016,7(36):58367-58380.
[17] ZHAO J,FANGZ,ZHA Z,et al.Quercetin inhibits cell viability,migration and invasion by regulating miR-16/HOXA10 axis in oral cancer[J].Eur J Pharmacol,2019,847:11-18.
[18] ZHOU J,GONG J,DING C,et al.Quercetin induces the apoptosis of human ovarian carcinoma cells by upregulating the expression of microRNA-145[J].Mol Med Rep,2015,12(2):3127-3131.
[19] ZHANG Z,LI B,XU P,et al.Integrated whole transcriptome profiling and bioinformatics analysis for revealing regulatory pathways associated with quercetin-induced apoptosis in HCT-116 cells[J].Front Pharmacol,2019,10:798.
[20] ZHANG X,GUO Q,CHEN J,et al.Quercetin enhances cisplatin sensitivity of human osteosarcoma cells by modulating microRNA-217-KRAS axis[J].Mol Cells,2015,38(7):638-642.
[21] ZANG X,CHENG M,ZHANG X,et al.Quercetin nanoformulations:a promising strategy for tumor therapy[J].Food Funct,2021,12(15):6664-6681.
[22] LOU M,ZHANG L N,JI P G,et al.Quercetin nanoparticles induced autophagy and apoptosis through AKT/ERK/Caspase-3 signaling pathway in human neuroglioma cells:in vitro and in vivo[J].Biomed Pharmacother,2016,84:1-9.
[23] RIVA A,RONCHI M,PETRANGOLINI G,et al.Improved oral absorption of quercetin from quercetin phytosome:a new delivery system based on food grade lecithin[J].Eur J Drug Metab Pharmacokinet,2019,44(2):169-177.
[24] 胡菲,车智慧,王哲,等.不同稳定剂修饰的槲皮素纳米晶在大鼠体内注射药动学及组织分布研究[J].中草药,2022,53(9):2697-2705.

备注/Memo

备注/Memo:
基金项目:浙江省教育厅一般科研项目(Y202249274)
△通信作者 E-mail:wuliangbang88@163.com
更新日期/Last Update: 2023-06-10