鼠李素下调Notch-1的表达对乳腺癌MCF-7细胞增殖、侵袭和迁移的调节作用
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摘要
目的探索鼠李素对乳腺癌MCF-7细胞增殖,侵袭和迁移的影响及其分子机制。方法MCF-7细胞分为4组:MCF-7(对照组)、鼠李素(1、2.5、5μM)组。CCK-8检测细胞增殖。Transwell检测细胞侵袭。划痕实验分析细胞迁移。蛋白印记检测Notch-1、CSL、Hes1、Ki67和VEGF表达。结果低浓度的(<5μM)的鼠李素不会影响MCF-7细胞活力;高浓度的(> 5μM)的鼠李素对MCF-7有显著毒性。2.5和5μM鼠李素组细胞增殖倍数明显低于对照组(P<0.05)。与对照组相比,鼠李素(1、2.5、5μM)组细胞侵袭和迁移明显降低(P <0.05)。另外,鼠李素(1、2.5、5μM)组Notch-1、CSL和Hes1相对蛋白表达量都明显少于对照组(P <0.05)。而且,Notch-1通路激活剂Jagged1(10μM)还可明显抑制鼠李素(5μM)诱导的Notch-1、CSL、Hes1、Ki67和VEGF表达的下降(P <0.05)。结论鼠李素通过下调Notch-1的表达抑制乳腺癌MCF-7细胞增殖,侵袭和迁移。
Objective To explore the effect, and its molecular mechanism, of rhamnetin on cell proliferation, invasion and migration of MCF-7 cell which is a kind of breast cancer cell. Methods MCF-7 cells were divided into four groups: MCF-7(control group) and rhamnetin(1, 2.5, 5 μM) groups. Cell proliferaion was detected by CCK-8. Cell invasion was measured by transwell. Cell migration was tested by wound healing. The expression of Notch-1, CSL, Hes1, Ki67 and VEGF were detected by western blot.Results Low concentration(< 5 μM) rhamnetin had no effect on the cell viability of MCF-7, while it was quite opposite under the circumstance of high concentration(> 5 μM) rhamnetin. The proliferation in rhamnetin(2.5, 5 μM) groups were lower than that of control group(P<0.05). Compared with control group,the invasion and migration in rhamnetin(1, 2.5, 5 μM) groups were decreased(P<0.05) with lower expression of Notch-1, CSL and Hes1 in rhamnetin(P<0.05). Moreover, the rhamnetin(5 μM)-induced declined expression of Notch-1, CSL, Hes1, Ki67 and VEGF were repressed by Notch-1 pathway activating agent Jagged1(10 μM). Conclusions Rhamnetin can restrain the proliferation, invasion and migration of MCF-7 via down-regulating the expression of Notch-1.
Objective To explore the effect, and its molecular mechanism, of rhamnetin on cell proliferation, invasion and migration of MCF-7 cell which is a kind of breast cancer cell. Methods MCF-7 cells were divided into four groups: MCF-7(control group) and rhamnetin(1, 2.5, 5 μM) groups. Cell proliferaion was detected by CCK-8. Cell invasion was measured by transwell. Cell migration was tested by wound healing. The expression of Notch-1, CSL, Hes1, Ki67 and VEGF were detected by western blot.Results Low concentration(< 5 μM) rhamnetin had no effect on the cell viability of MCF-7, while it was quite opposite under the circumstance of high concentration(> 5 μM) rhamnetin. The proliferation in rhamnetin(2.5, 5 μM) groups were lower than that of control group(P<0.05). Compared with control group,the invasion and migration in rhamnetin(1, 2.5, 5 μM) groups were decreased(P<0.05) with lower expression of Notch-1, CSL and Hes1 in rhamnetin(P<0.05). Moreover, the rhamnetin(5 μM)-induced declined expression of Notch-1, CSL, Hes1, Ki67 and VEGF were repressed by Notch-1 pathway activating agent Jagged1(10 μM). Conclusions Rhamnetin can restrain the proliferation, invasion and migration of MCF-7 via down-regulating the expression of Notch-1.
引文
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[2]Torre LA,Bray F,Siegel RL,et al.Global cancer statistics,2012[J]CA Cancer J Clin,2015,65(2):87-108.
[3]Magne Nde CB,Zingue S,Winter E,et al.Flavonoids,breast cancer chemopreventive and/or chemotherapeutic agents[J].Curr Med Chem,2015,22(30):3434-3446.
[4]Maggioni D,Biffi L,Nicolini G,et al.Flavonoids in oral cancer prevention and therapy[J].Eur J Cancer Prev,2015,24(6):517-528.
[5]Novo Belchor M,Hessel Gaeta H,Fabri Bittencourt Rodrigues C,et al.Evaluation of rhamnetin as an inhibitor of the pharmacological effect of secretory phospholipase A2[J].Molecules,2017,22(9).pii:E1441.
[6]Zhang W,Li B,Guo Y,et al.Rhamnetin attenuates cognitive deficit and inhibits hippocampal inflammatory response and oxidative stress in rats with traumatic brain injury[J].Cent Eur J Immunol,2015,40(1):35-41.
[7]Manu KA,Shanmugam MK,Ramachandran L,et al.Isorhamnetin augments the anti-tumor effect of capeciatbine through the negative regulation of NF-kappaB signaling cascade in gastric cancer[J].Cancer Lett,2015,363(1):28-36.
[8]Fan L,Strasser-Weippl K,Li JJ,et al.Breast cancer in China[J].Lancet Oncol,2014,15(7):e279-e289.
[9]钮红岺,黄晓萍,刘晓珑,等.阿司匹林对血小板诱导的乳腺癌MCF-7细胞上皮间质转化和迁移侵袭能力的影响[J].中国临床解剖学杂志,2017,35(2):183-187.
[10]Prasain JK,Rajbhandari R,Keeton AB,et al.Metabolism and growth inhibitory activity of cranberry derived flavonoids in bladder cancer cells[J].Food Funct,2016,7(9):4012-4019.
[11]Li Q,Ren FQ,Yang CL,et al.Anti-proliferation effects of isorhamnetin on lung cancer cells in vitro and in vivo[J].Asian Pac J Cancer Prev,2015,16(7):3035-3042.
[12]Hu S,Huang L,Meng L,et al.Isorhamnetin inhibits cell proliferation and induces apoptosis in breast cancer via Akt and mitogenactivated protein kinase kinase signaling pathways[J].Mol Med Rep,2015,12(5):6745-6751.
[13]Ramachandran L,Manu KA,Shanmugam MK,et al.Isorhamnetin inhibits proliferation and invasion and induces apoptosis through the modulation of peroxisome proliferator-activated receptor gamma activation pathway in gastric cancer[J].J Biol Chem,2012,287(45):38028-38040.
[14]Seo S,Seo K,Ki SH,et al.Isorhamnetin Inhibits Reactive Oxygen Species-Dependent Hypoxia Inducible Factor(HIF)-1alpha Accumulation[J].Biol Pharm Bull,2016,39(11):1830-1838.
[15]Li C,Yang D,Zhao Y,et al.Inhibitory effects of isorhamnetin on the invasion of human breast carcinoma cells by downregulating the expression and activity of matrix metalloproteinase-2/9[J].Nutr Cancer,2015,67(7):1191-1200.
[16]Gao XJ,Liu JW,Zhang QG,et al.Nobiletin inhibited hypoxia-induced epithelial-mesenchymal transition of lung cancer cells by inactivating of Notch-1 signaling and switching on miR-200b[J].Pharmazie,2015,70(4):256-262.
[17]Sun DW,Zhang HD,Mao L,et al.Luteolin inhibits breast cancer development and progression in vitro and in vivo by suppressing notch signaling and regulating miRNAs[J].Cell Physiol Biochem,2015,37(5):1693-1711.
[18]Kang J,Kim E,Kim W,et al.Rhamnetin and cirsiliol induce radiosensitization and inhibition of epithelial-mesenchymal transition(EMT)by miR-34a-mediated suppression of Notch-1 expression in nonsmall cell lung cancer cell lines[J].J Biol Chem,2013,288(38):27343-27357.
[19]El-Alfy TS,Ezzat SM,Hegazy AK,et al.Isolation of biologically active constituents from Moringa peregrina(Forssk.)Fiori.(family:Moringaceae)growing in Egypt[J].Pharmacogn Mag,2011,7(26):109-115.