Sphk1在舌鳞状细胞癌组织中的表达及其对舌癌细胞生物学行为的影响
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摘要
目的研究鞘氨醇激酶1(Sphk1)在舌鳞状细胞癌(TSCC)组织中的表达及意义,并探讨其对舌癌Tca8113P160细胞增殖、迁移及侵袭能力的影响。方法采用免疫组化法检测50例TSCC、28例口腔白斑病及10例癌旁正常舌黏膜标本中Sphk1的表达水平,分析其与TSCC患者临床病理特征的关系。将舌癌Tca8113P160细胞分为N,N-二甲基鞘氨醇(DMS)组和对照组,DMS组加入Sphk1抑制剂DMS,对照组加入无药物培养基。采用CCK-8法检测两组培养24、48、72 h的细胞增殖能力,Transwell实验检测细胞迁移和侵袭能力,Western blotting法检测Sphk1、N-cadherin、E-cadherin蛋白表达。结果 Sphk1在TSCC组织中的高表达率高于白斑及正常舌黏膜组织(P均<0.01)。Sphk1在TSCC组织中的高表达与肿瘤临床分期、淋巴结转移、T分期及组织学分级有关(P均<0.05)。DMS组培养24、48、72 h的细胞增殖OD值均低于对照组(P均<0.05)。DMS组在迁移实验和侵袭实验中的穿膜细胞数均较对照组减少(P均<0.01)。与对照组相比,DMS组Sphk1、N-cadherin蛋白表达减少,E-cadherin蛋白表达增加(P均<0.05)。结论 Sphk1在TSCC组织中高表达,参与TSCC的发生发展;其表达下调可抑制舌癌细胞的增殖、迁移及侵袭,可能与逆转上皮间质转化有关,Sphk1有望成为TSCC治疗的潜在靶点。
Objective To study the expression and significance of sphingosine kinase 1( Sph K1) in the tongue squamous cell carcinoma( TSCC) tissues and to observe the effects of Sph K1 on the proliferation,invasion,and migration of tongue cancer Tca8113 P160 cells. Methods Immunohistochemistry was used to detect the expression of Sphk1 in TSCC( 50 cases),leukoplakia( 28 cases),and normal tongue mucosa( 10 cases) specimens. The correlation between Sphk1 expression and clinical features was analyzed. Tca8113 P160 cells were divided into two groups: the N,N-dimethylsphingosine( DMS) group which was treated with Sphk1 inhibitor DMS,and the control group which was added with medium with no medication. The proliferation of cells was detected by CCK-8 at 24,48,and 72 h. The migration and invasion of cells were detected by Transwell assay. The protein expression of Sph K1,N-cadherin,and E-cadherin was detected by Western blotting. Results The high expression rate of Sphk1 in TSCC tissues was higher than that in leukoplakia and normal mucosa tissues( all P < 0. 01),which was correlated with clinical stage,lymph node metastasis,T stage,and histological differentiation( all P < 0. 05). After incubation for 24,48,and 72 h,the OD value of DMS group was lower than that in the control group( all P < 0. 05). The migration and invasion assay showed the number of transmembrane cells in the DMS group was significantly less than that in the control group( P < 0. 01). The expression of Sphk1 and N-cadherin of the DMS group was lower,while the expression of E-cadherin protein was higher than that in the control group( all P <0. 05). Conclusions Sphk1 is highly expressed in TSCC,and the down-regulation of Sphk1 can inhibit the proliferation,invasion,and migration of Tca8113 P160 cells and reverse the epithelial-mesenchymal transition. Sphk1 may be a potentialtarget for TSCC treatment.
Objective To study the expression and significance of sphingosine kinase 1( Sph K1) in the tongue squamous cell carcinoma( TSCC) tissues and to observe the effects of Sph K1 on the proliferation,invasion,and migration of tongue cancer Tca8113 P160 cells. Methods Immunohistochemistry was used to detect the expression of Sphk1 in TSCC( 50 cases),leukoplakia( 28 cases),and normal tongue mucosa( 10 cases) specimens. The correlation between Sphk1 expression and clinical features was analyzed. Tca8113 P160 cells were divided into two groups: the N,N-dimethylsphingosine( DMS) group which was treated with Sphk1 inhibitor DMS,and the control group which was added with medium with no medication. The proliferation of cells was detected by CCK-8 at 24,48,and 72 h. The migration and invasion of cells were detected by Transwell assay. The protein expression of Sph K1,N-cadherin,and E-cadherin was detected by Western blotting. Results The high expression rate of Sphk1 in TSCC tissues was higher than that in leukoplakia and normal mucosa tissues( all P < 0. 01),which was correlated with clinical stage,lymph node metastasis,T stage,and histological differentiation( all P < 0. 05). After incubation for 24,48,and 72 h,the OD value of DMS group was lower than that in the control group( all P < 0. 05). The migration and invasion assay showed the number of transmembrane cells in the DMS group was significantly less than that in the control group( P < 0. 01). The expression of Sphk1 and N-cadherin of the DMS group was lower,while the expression of E-cadherin protein was higher than that in the control group( all P <0. 05). Conclusions Sphk1 is highly expressed in TSCC,and the down-regulation of Sphk1 can inhibit the proliferation,invasion,and migration of Tca8113 P160 cells and reverse the epithelial-mesenchymal transition. Sphk1 may be a potentialtarget for TSCC treatment.
引文
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[14]徐香兰,韩冰,钟启平,等.RNA干扰沉默HIF-1α基因对舌癌Tca8113细胞侵袭和迁移的影响[J].山东医药,2013,53(43):5-8.
[15]Liu H,Ma Y,He HW,et al.SPHK1(sphingosine kinase 1)induces epithelial-mesenchymal transition by promoting the autophagy-linked lysosomal degradation of CDH1/E-cadherin in hepatoma cells[J].Autophagy,2017,13(5):900-913.
[16]诸葛春风,刘诗权,谭林,等.Sphk1和FAK对人结肠癌HCT116细胞上皮间质转化的影响[J].中国病理生理杂志,2016,32(3):439-444.
[2]Siegel R,Naishadham D,Jemal A.Cancer statistics,2013[J].CA Cancer J Clin,2013,63(1):11-30.
[3]Pitson SM,Powell JA,Bonder CS.Regulation of sphingosine kinase in hematological malignancies and other cancers[J].Anticancer Agents Med Chem,2011,11(9):799-809.
[4]Heernan-Stroud LA,Obeid LM.Sphingosine kinase 1 in cancer[J].Adv Cancer Res,2013,117:201-235.
[5]张彩霞,何红伟,邵荣光.鞘氨醇激酶与肿瘤[J].药学学报,2013,48(7):971-978.
[6]Xia P,Gamble JR,Wang L,et al.An oncogenic role of sphingosine kinase[J].Curr Biol,2000,10(23):1527-1530.
[7]Zheng XD,Zhang Y,Qi XW,et al.Role of Sphk1 in the malignant transformation of breast epithelial cells and breast cancer progression[J].Indian J Cancer,2014,51(4):524.
[8]Shirai K,Kaneshiro T,Wada M,et al.A role of sphingosine kinase 1 in head and neck carcinogenesis[J].Cancer Prev Res,2011,4(3):454-462.
[9]Liu SQ,Su YJ,Qin MB,et al.Sphingosine kinase 1 promotes tumor progression and confers malignancy phenotypes of colon cancer by regulating the focal adhesion kinase pathway and adhesion molecules[J].Int J Oncol,2012,42(2):617-626.
[10]Li W,Tian Z,Qin H,et al.High expression of sphingosine kinase1 is associated with poor prognosis in nasopharyngeal carcinoma[J].Biochem Biophys Res Commun,2015,460(2):341-347.
[11]Song L,Xiong H,Li J,et al.Sphingosine kinase-1 enhances resistance to apoptosis through activation of PI3K/Akt/NF-κB pathway in human non-small cell lung cancer[J].Clin Cancer Res,2011,17(7):1839-1849.
[12]Zhao Y,Ling ZQ,Hao YB,et al.MiR-124 acts as a tumor suppressor by inhibiting the expression of sphingosine kinase 1 and its downstream signaling in head and neck squamous cell carcinoma[J].Oncotarget,2017,8(15):25005-25020.
[13]Pan J,Tao YF,Zhou Z,et al.An novel role of sphingosine kinase-1(SPHK1)in the invasion and metastasis of esophageal carcinoma[J].J Transl Med,2011,9:157.
[14]徐香兰,韩冰,钟启平,等.RNA干扰沉默HIF-1α基因对舌癌Tca8113细胞侵袭和迁移的影响[J].山东医药,2013,53(43):5-8.
[15]Liu H,Ma Y,He HW,et al.SPHK1(sphingosine kinase 1)induces epithelial-mesenchymal transition by promoting the autophagy-linked lysosomal degradation of CDH1/E-cadherin in hepatoma cells[J].Autophagy,2017,13(5):900-913.
[16]诸葛春风,刘诗权,谭林,等.Sphk1和FAK对人结肠癌HCT116细胞上皮间质转化的影响[J].中国病理生理杂志,2016,32(3):439-444.