白杨素调控miR-320和Twist1表达抑制卵巢癌干细胞样细胞体外致瘤功能
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摘要
目的 :研究白杨素能否调节miR-320和Twist1表达抑制人卵巢癌SKOV3源性干细胞样细胞体外致瘤能力。方法 :不同浓度白杨素(5.0、10.0和20.0μM)处理肿瘤球培养得到的第2代球细胞即卵巢癌干细胞样细胞。实时定量PCR检测miR-320表达水平。球形成率测定和琼脂集落形成试验评价体外致瘤能力。Western blot分析Twist1蛋白表达。分别用miR-320模拟物或Twist1基因转导探讨白杨素的作用机制。结果 :与亲本细胞相比,SKOV3源性卵巢癌干细胞样细胞miR-320低表达,而Twist1蛋白高表达。白杨素(5、10和20μM)以浓度依赖方式降低SKOV3源性卵巢癌干细胞样细胞球形成率和集落形成率;同时,上调miR-320表达和下调Twist1蛋白表达。miR-320模拟物转染增强白杨素(10μM)的效应。Twist1基因转导能有效逆转白杨素(10μM)的作用。结论 :白杨素通过调控miR-320/Twist1信号轴抑制SKOV3源性卵巢癌干细胞样细胞体外致瘤性。
Objective To investigate whether chrysin can inhibit in vitro tumorigenicity of ovarian cancer stem-like cells(OCSLCs) derived from human ovarian cancer SKOV3 cell line by modulating the expression of Mi R-320 and Twist1. MethodsSKOV3-derived OCSLCs were obtained from the second generation of cancer cells using tumor spheroid formation culture, which were treated with various concentrations(5.0, 10.0 and 20.0μM) of chrysin. Real-time quantitative PCR was used to detect the expression of Mi R-320. Ttumorigenicity in vitro were evaluated by Spheroid and colony formation assay. Western blot was used to examine Twist1 protein expression. Mi R-320 mimics transfection or Twist1 gene transduction were used to elucidate the mechanism underlying the action of chrysin in OCSLCs. Results The miR-320 of SKOV3-derived OCSLCs was low-expressed and Twist1 protein was highly expressed, compared to the parental cells. Chrysin(5, 10 and 20μM) decreased spheroid and colony forming rates of SKOV3-derived OCSLCs in a concentration-dependent manner, while up-regulated the expression of miR-320 and down-regulated Twist1. miR-320 mimics transfection enhances the effect of chrysin(10μM). Twist1 gene transduction can effectively reverse the effect of chrysin(10μM). Conclusion Chrysin inhibits in vitro tumorigenicity of SKOV3-derived OCSLCs by regulating the miR-320/Twist1 axis.
Objective To investigate whether chrysin can inhibit in vitro tumorigenicity of ovarian cancer stem-like cells(OCSLCs) derived from human ovarian cancer SKOV3 cell line by modulating the expression of Mi R-320 and Twist1. MethodsSKOV3-derived OCSLCs were obtained from the second generation of cancer cells using tumor spheroid formation culture, which were treated with various concentrations(5.0, 10.0 and 20.0μM) of chrysin. Real-time quantitative PCR was used to detect the expression of Mi R-320. Ttumorigenicity in vitro were evaluated by Spheroid and colony formation assay. Western blot was used to examine Twist1 protein expression. Mi R-320 mimics transfection or Twist1 gene transduction were used to elucidate the mechanism underlying the action of chrysin in OCSLCs. Results The miR-320 of SKOV3-derived OCSLCs was low-expressed and Twist1 protein was highly expressed, compared to the parental cells. Chrysin(5, 10 and 20μM) decreased spheroid and colony forming rates of SKOV3-derived OCSLCs in a concentration-dependent manner, while up-regulated the expression of miR-320 and down-regulated Twist1. miR-320 mimics transfection enhances the effect of chrysin(10μM). Twist1 gene transduction can effectively reverse the effect of chrysin(10μM). Conclusion Chrysin inhibits in vitro tumorigenicity of SKOV3-derived OCSLCs by regulating the miR-320/Twist1 axis.
引文
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[12]He X,Wei Y,Wang Y,et al.Mi R-381 functions as a tumor suppressor in colorectal cancer by targeting Twist1[J].Onco Targets Ther,2016,9:1231-9.
[13]Li CW,Xia W,Lim SO,et al.AKT1 Inhibits Epithelial-to-Mesenchymal Transition in Breast Cancer through Phosphorylation-Dependent Twist1 Degradation[J].Cancer Res,2016,76(6):1451-62.
[14]Wu K,Wang B,Chen Y,et al.DAB2IP regulates the chemoresistance to pirarubicin and tumor recurrence of non-muscle invasive bladder cancer through STAT3/Twist1/P-glycoprotein signaling[J].Cell Signal,2015,27(12):2515-23.
[15]Zhu X,Shen H,Yin X,et al.miR-186 regulation of Twist1 and ovarian cancer sensitivity to cisplatin[J].Oncogene,2016,35(3):323-32.
[2]Zhao XC,Cao XC,Liu F,et al.Regulation of the FOXO3a/Bim signaling pathway by 5,7-dihydroxy-8-nitrochrysin in MDA-MB-453 breast cancer cells[J].Oncol Lett,2013,5(3):929-934.
[3]Yang JF,Cao JG,Tian L,et al.5,7-Dimethoxyflavone sensitizes TRAIL-induced apoptosis through DR5 upregulation in hepatocellular carcinoma cells[J].Cancer Chemother Pharmacol,2012,69(1):195-206.
[4]李华珍,陈艳红,方银兰,等.白杨素对SKOV3细胞系干样细胞球形成和CK2α表达的影响[J].中华医学杂志,2016,96(25):2013-2016.
[5]董巍檑,宁映霞,陈阿,等.TWIST1基因敲低对DFOG抑制OVCAR3源性球细胞自我更新和Fox M1表达的影响[J].湖南师范大学学报(医学版),2017,14(05):4-7.
[6]Ren KQ,Cao XZ,Liu ZH,et al.8-bromo-5-hydroxy-7-methoxychrysin targeting for inhibition of the properties of liver cancer stem cells by modulation of Twist signaling[J].Int J Oncol,2013,43(5):1719-29.
[7]He M,Cao XC,He GC,et al.Casticin inhibits epithelial-mesenchymal transition of liver cancer stem cells of the SMMC-7721 cell line through downregulating Twist[J].Oncol Lett,2014,7(5):1625-1631.
[8]Li C,Duan P,Wang J,et al.miR-320 inhibited ovarian cancer oncogenicity via targeting TWIST1 expression[J].Am J Transl Res,2017,9(8):3705-3713.
[9]Wang SC,Lin XL,Li J,et al.Micro RNA-122 triggers mesenchymalepithelial transition and suppresses hepatocellular carcinoma cell motility and invasion by targeting Rho A[J].PLo S One,2014,9(7):e101330.
[10]Lei T,Zhu Y,Jiang C,et al.Micro RNA-320 was downregulated in nonsmall cell lung cancer and inhibited cell proliferation,migration and invasion by targeting fatty acid synthase[J].Mol Med Rep,2016,14(2):1255-62.
[11]Vishnubalaji R,Hamam R,Yue S,et al.Micro RNA-320 suppresses colorectal cancer by targeting SOX4,FOXM1,and FOXQ1[J].Oncotarget,2016,7(24):35789-35802.
[12]He X,Wei Y,Wang Y,et al.Mi R-381 functions as a tumor suppressor in colorectal cancer by targeting Twist1[J].Onco Targets Ther,2016,9:1231-9.
[13]Li CW,Xia W,Lim SO,et al.AKT1 Inhibits Epithelial-to-Mesenchymal Transition in Breast Cancer through Phosphorylation-Dependent Twist1 Degradation[J].Cancer Res,2016,76(6):1451-62.
[14]Wu K,Wang B,Chen Y,et al.DAB2IP regulates the chemoresistance to pirarubicin and tumor recurrence of non-muscle invasive bladder cancer through STAT3/Twist1/P-glycoprotein signaling[J].Cell Signal,2015,27(12):2515-23.
[15]Zhu X,Shen H,Yin X,et al.miR-186 regulation of Twist1 and ovarian cancer sensitivity to cisplatin[J].Oncogene,2016,35(3):323-32.