贝母素乙调控PI3K/Akt/mTOR通路减缓上皮-间质转化进程抑制人肺癌A549细胞侵袭及迁移的研究
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摘要
目的探讨贝母素乙对人肺癌A549细胞侵袭及迁移的影响。方法采用终浓度为0、50、100、200μmol/L贝母素乙处理A549细胞,通过细胞侵袭实验、细胞划痕实验、实时荧光定量PCR(q RT-PCR)法、ELISA法及Western blotting实验检测贝母素乙对A549细胞侵袭、迁移的影响及潜在机制。结果与对照组比较,贝母素乙各浓度组的A549细胞穿膜数、细胞间划痕的愈合率及MMP-9、MMP-2表达显著下降(P<0.01);与对照组比较,贝母素乙各浓度组的A549细胞E-cadherin m RNA表达明显增加(P<0.01),而N-cadherin和vimentin mRNA表达均显著降低(P<0.01);与对照组比较,除50μmol/L贝母素乙处理A549细胞24h后的FN蛋白表达无显著性变化外(P>0.05),其余贝母素乙各浓度组在各时间点上的FN蛋白表达均显著下降(P<0.05、0.01);与对照组比较,贝母素乙各浓度组的p-PI3K/PI3K、p-mTOR/mTOR值及100、200μmol/L贝母素乙组的p-Akt/Akt值显著下降(P<0.05、0.01)。结论贝母素乙具有抑制A549细胞侵袭及迁移能力的作用,该作用与贝母素乙调控PI3K/Akt/mTOR通路活性进而减缓上皮-间质转化(EMT)进程有关。
Objective To detect the influence of peiminine on the invasion and migration of human lung cancer A549 cells. Methods A549 cells were treated with peiminine at the final concentrations of 0, 50, 100, and 200 μmol/L, respectively. The influence of peiminine on the invasion and migration of A549 cells and its underlying mechanisms were investigated by cell invasion experiment,cell scratch experiment, real-time quantitative PCR, ELISA, and Western blotting. Results Compared with 0 μmol/L peiminine group,the cell transmembrane number and scratch wound healing rate and expressions of MMP-9 and MMP-2 in the group treated with different concentrations of peiminine significantly decreased(P < 0.01). Compared with 0 μmol/L peiminine group, the m RNA expression of E-cadherin increased significantly(P < 0.01), while the m RNA expressions of N-cadherin and vimentin decreased significantly(P < 0.01). Compared with 0 μmol/L peiminine group, FN protein expression was significantly decreased in all the groups with different concentrations of peiminine group except treatment with 50 μmol/L peiminine after 24 h(P < 0.05, 0.01). Compared with0 μmol/L peiminine group, the ratio of p-PI3 K/PI3 K and p-mTOR/mTOR in all concentrations of peiminine groups and p-Akt/Akt in100 and 200 peiminine groups were significantly decreased(P < 0.05, 0.01). Conclusion Peiminine can inhibit the invasion and migration of A549 cells, which may be related to the activation of PI3 K/Akt/mTOR pathway and decreasing the epithelialmesenchymal transition process.
Objective To detect the influence of peiminine on the invasion and migration of human lung cancer A549 cells. Methods A549 cells were treated with peiminine at the final concentrations of 0, 50, 100, and 200 μmol/L, respectively. The influence of peiminine on the invasion and migration of A549 cells and its underlying mechanisms were investigated by cell invasion experiment,cell scratch experiment, real-time quantitative PCR, ELISA, and Western blotting. Results Compared with 0 μmol/L peiminine group,the cell transmembrane number and scratch wound healing rate and expressions of MMP-9 and MMP-2 in the group treated with different concentrations of peiminine significantly decreased(P < 0.01). Compared with 0 μmol/L peiminine group, the m RNA expression of E-cadherin increased significantly(P < 0.01), while the m RNA expressions of N-cadherin and vimentin decreased significantly(P < 0.01). Compared with 0 μmol/L peiminine group, FN protein expression was significantly decreased in all the groups with different concentrations of peiminine group except treatment with 50 μmol/L peiminine after 24 h(P < 0.05, 0.01). Compared with0 μmol/L peiminine group, the ratio of p-PI3 K/PI3 K and p-mTOR/mTOR in all concentrations of peiminine groups and p-Akt/Akt in100 and 200 peiminine groups were significantly decreased(P < 0.05, 0.01). Conclusion Peiminine can inhibit the invasion and migration of A549 cells, which may be related to the activation of PI3 K/Akt/mTOR pathway and decreasing the epithelialmesenchymal transition process.
引文
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[20]姜爽,任艳平,魏琳,等.人参皂苷CK对MCF-7细胞增殖、凋亡、上皮间质转化、PI3K/Akt信号通路的影响[J].中成药,2018,40(9):1925-1929.
[21]Baek S H,Ko J H,Lee J H,et al.Ginkgolic acid inhibits invasion and migration and TGF-β-Induced EMT of lung cancer cells through PI3K/Akt/mTOR inactivation[J].JCell Physiol,2017,232(2):346-354.
[22]Feng L M,Wang X F,Huang Q X.Thymoquinone induces cytotoxicity and reprogramming of EMT in gastric cancer cells by targeting PI3K/Akt/mTORpathway[J].J Biosci,2017,42(4):547-554.
[2]Okamoto T,Yano T,Shimokawa M,et al.A phase IIrandomized trial of adjuvant chemotherapy with S-1versus S-1 plus cisplatin for completely resected pathological stage II/IIIA non-small cell lung cancer[J].Lung Cancer,2018,124(1):255-259.
[3]Liao Y H,Li C I,Lin C C,et al.Traditional Chinese medicine as adjunctive therapy improves the long-term survival of lung cancer patients[J].J Cancer Res Clin Oncol,2017,143(12):2425-2435.
[4]Li L,Wang S,Yang X,et al.Traditional Chinese medicine,Fuzheng Kang-Ai decoction,inhibits metastasis of lung cancer cells through the STAT3/MMP9pathway[J].Mol Med Rep,2017,16(3):2461-2468.
[5]丁常宏,郭盛磊,孙海峰,等.药用植物平贝母的研究进展[J].中医药导报,2018,24(3):73-75.
[6]Pan F,Hou K,Gao F,et al.Peimisine and peiminine production by endophytic fungus Fusarium sp.isolated from Fritillaria unibracteata var.wabensis[J].Phytomedicine,2014,21(8/9):1104-1109.
[7]Zheng Z,Xu L,Zhang S,et al.Peiminine inhibits colorectal cancer cell proliferation by inducing apoptosis and autophagy and modulating key metabolic pathways[J].Oncotarget,2017,8(29):47619-47631.
[8]张喆,何勤思,吴晨雯,等.中药提取物贝母素乙对人结肠癌HCT-116细胞基因表达的影响[J].中医杂志,2016,57(17):1504-1509.
[9]Tang Q,Wang Y,Ma L,et al.Peiminine serves as an adriamycin chemosensitizer in gastric cancer by modulating the EGFR/FAK pathway[J].Oncol Rep,2018,39(3):1299-1305.
[10]王云飞,顾政一,聂勇战,等.贝母素乙增强阿霉素对胃癌多药耐药裸鼠移植瘤的抑制作用及其机制研究[J].中草药,2014,45(5):686-690.
[11]孙师钢,杜佳蕾,孙晓琦,等.人参皂苷CK抑制乳腺癌MCF-7细胞迁移和侵袭的实验研究[J].时珍国医国药,2018,29(5):1068-1070.
[12]Teoh-Fitzgerald M L,Fitzgerald M P,Jensen T J,et al.Genetic and epigenetic inactivation of extracellular superoxide dismutase promotes an invasive phenotype in human lung cancer by disrupting ECM homeostasis[J].Mol Cancer Res,2012,10(1):40-51.
[13]Guo B,Zhang J,Li Q,et al.Hypermethylation of mi R-338-3p and Impact of its suppression on cell metastasis through N-cadherin accumulation at the cell-cell junction and degradation of MMP in gastric cancer[J].Cell Physiol Biochem,2018,50(2):411-425.
[14]谢珂,郑卫红,谭潇.KLKs促进肿瘤增殖、迁移与侵袭的分子机制[J].中国生物化学与分子生物学报,2018,34(9):935-941.
[15]Kim C W,Hwang K A,Choi K C.Anti-metastatic potential of resveratrol and its metabolites by the inhibition of epithelial-mesenchymal transition,migration,and invasion of malignant cancer cells[J].Phytomedicine,2016,23(14):1787-1796.
[16]朱宏明,尹丽,何侠.Notch信号通路介导EMT与肿瘤侵袭转移的研究进展[J].肿瘤学杂志,2018,24(8):808-812.
[17]Topalovski M,Brekken R A.Matrix control of pancreatic cancer:New insights into fibronectin signaling[J].Cancer Lett,2016,381(1):252-258.
[18]Qin S,Zhang B,Xiao G,et al.Fibronectin protects lung cancer cells against docetaxel-induced apoptosis by promoting Src and caspase-8 phosphorylation[J].Tumour Biol,2016,37(10):13509-13520.
[19]Guerrero-Zotano A,Mayer I A,Arteaga C L.PI3K/AKT/mTOR:Role in breast cancer progression,drug resistance,and treatment[J].Cancer Metastasis Rev,2016,35(4):515-524.
[20]姜爽,任艳平,魏琳,等.人参皂苷CK对MCF-7细胞增殖、凋亡、上皮间质转化、PI3K/Akt信号通路的影响[J].中成药,2018,40(9):1925-1929.
[21]Baek S H,Ko J H,Lee J H,et al.Ginkgolic acid inhibits invasion and migration and TGF-β-Induced EMT of lung cancer cells through PI3K/Akt/mTOR inactivation[J].JCell Physiol,2017,232(2):346-354.
[22]Feng L M,Wang X F,Huang Q X.Thymoquinone induces cytotoxicity and reprogramming of EMT in gastric cancer cells by targeting PI3K/Akt/mTORpathway[J].J Biosci,2017,42(4):547-554.