长期低氧条件下LOX促进胃癌转移的机制
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摘要
目的探讨赖氨酰氧化酶(LOX)促进低氧耐性胃癌(HRGC)细胞转移的机制。方法胃癌细胞系MGC803和BGC823常规培养,HRGC细胞系MGC803/hypo和BGC823/hypo培养于2%O_2条件下。Transwell法检测各胃癌细胞迁移能力差异;PCR检测各胃癌细胞LOX mRNA水平;siRNA瞬时沉默LOX表达;Western blot法检测各胃癌细胞LOX、AKT等表达变化及AKT通路活化情况。结果与亲本胃癌细胞比较,HRGC细胞的迁移能力显著增强(P <0.05),且LOX mRNA和蛋白表达水平明显增加(P <0.05);瞬时沉默LOX后,HRGC细胞的迁移能力显著下降(P <0.05)。与亲本胃癌细胞比较,HRGC细胞中AKT磷酸化水平明显增加;瞬时沉默LOX明显降低了HRGC细胞中的AKT磷酸化水平(P <0.05)。结论长期低氧条件下LOX上调,并通过活化AKT通路促进胃癌细胞转移。
Objective To investigate the mechanism of lysyl oxidase(LOX) on metastasis promotion in hypoxia-resistant gastric cancer(HRGC) cells. Methods The gastric cancer cell lines, MGC803 and BGC823 were routinely cultured.HRGC cell lines, MGC803/hypo and BGC823/hypo were cultured under 2%O_2 conditions. The migration ability of gastric cancer cells was detected by Transwell assay. LOX mRNA level was detected by PCR; LOX siRNA was used for LOX transiently silencing; Western blot was used to detect the protein level of LOX, AKT and the activation of AKT pathways. Results Compared with the parental gastric cancer cells, the migration ability of HRGC cells was significantly enhanced(P < 0.05), the expression level of mRNA and protein of LOX were significantly increased(P < 0.05). After transient silencing of LOX, the migration ability was significantly decreased in HRGC cells(P < 0.05). Compared with parental gastric cancer cells, AKT phosphorylation was significantly increased(P < 0.05), whereas AKT phosphorylation was significantly decreased after transient silencing of LOX in HRGC cells(P < 0.05). Conclusion Long-term hypoxic conditions increase LOX and promote gastric cancer cell metastasis by activating the AKT pathway.
Objective To investigate the mechanism of lysyl oxidase(LOX) on metastasis promotion in hypoxia-resistant gastric cancer(HRGC) cells. Methods The gastric cancer cell lines, MGC803 and BGC823 were routinely cultured.HRGC cell lines, MGC803/hypo and BGC823/hypo were cultured under 2%O_2 conditions. The migration ability of gastric cancer cells was detected by Transwell assay. LOX mRNA level was detected by PCR; LOX siRNA was used for LOX transiently silencing; Western blot was used to detect the protein level of LOX, AKT and the activation of AKT pathways. Results Compared with the parental gastric cancer cells, the migration ability of HRGC cells was significantly enhanced(P < 0.05), the expression level of mRNA and protein of LOX were significantly increased(P < 0.05). After transient silencing of LOX, the migration ability was significantly decreased in HRGC cells(P < 0.05). Compared with parental gastric cancer cells, AKT phosphorylation was significantly increased(P < 0.05), whereas AKT phosphorylation was significantly decreased after transient silencing of LOX in HRGC cells(P < 0.05). Conclusion Long-term hypoxic conditions increase LOX and promote gastric cancer cell metastasis by activating the AKT pathway.
引文
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[19]Lu H,Jiang T,Ren K,et al.RUNX2 Plays An Oncogenic Role in Esophageal Carcinoma by Activating the PI3K/AKT and ERK Signaling Pathways[J].Cell Physiol Biochem,2018,49(1):217-225.
[20]Shi L,Zhang N,Liu H,et al.Lysyl oxidase inhibition viaβ-aminoproprionitrile hampers human umbilical vein endothelial cell angiogenesis and migration in vitro[J].Mol Med Rep,2018,17(4):5029-5036.
[2]Bertout JA,Patel SA,Simon MC.The impact of O2availability on human cancer[J].Nat Rev Cancer,2008,8(12):967-975.
[3]Kasashima H,Yashiro M,Kinoshita H,et al.Lysyl oxidase is associated with the epithelial-mesenchymal transition of gastric cancer cells in hypoxia[J].Gastric Cancer,2016,19(2):431-442.
[4]Johnston KA,Lopez KM.Lysyl oxidase in cancer inhibition and metastasis[J].Cancer Lett,2018,417:174-181.
[5]Xie Q,Xie J,Tian T,et al.Hypoxia triggers angiogenesis by increasing expression of LOX genes in 3-D culture of ASCs and ECs[J].Exp Cell Res,2017,352(1):157-163.
[6]Balkwill FR,Capasso M,Hagemann T.The tumor microenvironment at a glance[J].J Cell SCI,2012,125(23):5591-5596.
[7]Mazure NM.Hypoxia signalling in cancer and approaches to enforce tumour regression[J].Nature,2006,441(7092):437-443.
[8]Cox TR,Gartland A,Erler JT.Lysyl Oxidase,a Targetable Secreted Molecule Involved in Cancer Metastasis[J].Cancer Res,2016,76(2):188-192.
[9]Erler JT,Bennewith KL,Cox TR,et al.Hypoxia-induced lysyl oxidase is a critical mediator of bone marrow cell recruitment to form the premetastatic niche[J].Cancer Cell,2009,15(1):35-44.
[10]Baker AM,Cox TR,Bird D,et al.The Role of Lysyl Oxidase in SRC-Dependent Proliferation and Metastasis of Colorectal Cancer[J].J Natl Cancer I,2011,103(5):407-424.
[11]Baker AM,Bird D,Lang G,et al.Lysyl oxidase enzymatic function increases stiffness to drive colorectal cancer progression through FAK[J].Oncogene,2016,35(49):6341-6349.
[12]El-Haibi CP,Bell GW,Zhang J,et al.Critical role for lysyl oxidase in mesenchymal stem cell-driven breast cancer malignancy[J].Proc Natl Acad Sci U S A,2012,109(43):17460-17465.
[13]Huang SP,Chiou J,Jan YH,et al.Over-expression of lysyl oxidase is associated with poor prognosis and response to therapy of patients with lower grade gliomas[J].Biochem Biophys Res Commun,2018,501(3):619-627.
[14]Hay N,Sonenberg N.Upstream and downstream of m TOR[J].Gene Dev,2004,18(16):1926-1945.
[15]Hamzehzadeh L,Atkin SL,Majeed M,et al.The versatile role of curcumin in cancer prevention and treatment:Afocus on PI3K/AKT pathway[J].J Cell Physiol,2018,233(10):6530-6537.
[16]Fukui H,Zhang X,Sun C,et al.IL-22 produced by cancer-associated fibroblasts promotes gastric cancer cell invasion via STAT3 and ERK signaling[J].Br J Cancer,2014,111(4):763-771.
[17]Bahrami A,Hassanian SM,Shahidsales S,et al.Targeting the RAS Signaling Pathway as a Potential Therapeutic Target in the Treatment of Colorectal Cancer[J].J Cell Physiol,2018,233(3):2058-2066.
[18]D′Amico S,Shi J,Martin BL,et al.STAT3 is a master regulator of epithelial identity and KRAS-driven tumorigenesis[J].Genes Dev,2018,32(17/18):1175-1187
[19]Lu H,Jiang T,Ren K,et al.RUNX2 Plays An Oncogenic Role in Esophageal Carcinoma by Activating the PI3K/AKT and ERK Signaling Pathways[J].Cell Physiol Biochem,2018,49(1):217-225.
[20]Shi L,Zhang N,Liu H,et al.Lysyl oxidase inhibition viaβ-aminoproprionitrile hampers human umbilical vein endothelial cell angiogenesis and migration in vitro[J].Mol Med Rep,2018,17(4):5029-5036.