RNA干扰FAK表达抑制宫颈癌细胞侵袭、迁移能力的研究
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摘要
目的探讨黏着斑激酶(FAK)表达量降低对宫颈癌细胞侵袭、迁移能力的影响以及机制研究。方法以宫颈癌He La细胞为研究对象,转染RNA干扰载体,实验分3组:空白对照组(未做任何处理的细胞)、阴性对照组(细胞转染对照载体FAK-p Genesil-Ctrl)、干扰组(细胞转染重组质粒FAK-pGenesil-siRNA)。Transwell法检测细胞的迁移、侵袭能力,Western blot法检测细胞中的FAK、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)、上皮细胞钙黏蛋白(E-cadherin)的蛋白表达量。结果干扰组细胞中FAK蛋白的表达量低于空白对照组和阴性对照组(P﹤0.05);干扰组细胞的迁移、侵袭数少于空白对照组和阴性对照组(P﹤0.05);干扰组细胞中MMP-2、MMP-9蛋白的表达量低于空白对照组和阴性对照组,但E-cadherin蛋白的表达量高于空白对照组和阴性对照组(P﹤0.05)。结论 RNA干扰FAK的表达量可以降低宫颈癌He La细胞的侵袭、迁移能力,其作用机制可能是通过影响细胞上皮间质转化相关因子MMP-2、MMP-9、E-cadherin蛋白的表达量。
Objective To investigate the effect and mechanism of FAK on the invasion and migration of cervical cancer cells. Method Cervical cancer cells He La were used in the study, the cells were transfected with RNAi interfering vector, and then were divided as 3 groups: blank control group(untreated cells); negative control group(transfected with FAK-p Genesil-Ctrl vector); interference group(transfected with recombinant plasmid FAK-p Genesil-siRNA). Transwell was applied to determine the migration and invasion of cells, while the expression of FAK, MMP-2, MMP-9 and E-cadherin protein in the cells were detected by Western blot. Result The expression of FAK protein in the interference group was significantly lower than that in the blank control group and the negative control group(P<0.05); lower ability of migration and invasion of the cells was observed in interference group than in the blank control group and the negative control group(P<0.05); the expression of MMP-2 and MMP-9 in interference group were significantly lower than that of the control group and the negative control group, but the expression of E-cadherin was significantly higher than that of blank control group and negative control group(P<0.05). Conclusion The expression of FAK interfered by RNA may lead to decreased ability of invasion, and migration of cervical cancer cells He La, which be related to the expression epithelialmesenchymal transformation-related factors, including MMP-2, MMP-9 and E-cadherin protein.
Objective To investigate the effect and mechanism of FAK on the invasion and migration of cervical cancer cells. Method Cervical cancer cells He La were used in the study, the cells were transfected with RNAi interfering vector, and then were divided as 3 groups: blank control group(untreated cells); negative control group(transfected with FAK-p Genesil-Ctrl vector); interference group(transfected with recombinant plasmid FAK-p Genesil-siRNA). Transwell was applied to determine the migration and invasion of cells, while the expression of FAK, MMP-2, MMP-9 and E-cadherin protein in the cells were detected by Western blot. Result The expression of FAK protein in the interference group was significantly lower than that in the blank control group and the negative control group(P<0.05); lower ability of migration and invasion of the cells was observed in interference group than in the blank control group and the negative control group(P<0.05); the expression of MMP-2 and MMP-9 in interference group were significantly lower than that of the control group and the negative control group, but the expression of E-cadherin was significantly higher than that of blank control group and negative control group(P<0.05). Conclusion The expression of FAK interfered by RNA may lead to decreased ability of invasion, and migration of cervical cancer cells He La, which be related to the expression epithelialmesenchymal transformation-related factors, including MMP-2, MMP-9 and E-cadherin protein.
引文
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[13]O’Brien S,Golubovskaya VM,Conroy J,et al.FAK inhibition with small molecule inhibitor Y15 decreases viability,clonogenicity,and cell attachment in thyroid cancer cell lines and synergizes with targeted therapeutics[J].Oncotarget,2014,5(17):7945-7959.
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[15]郝臻凤,王娟,周留林,等.FAK在宫颈癌中的表达及临床意义[J].扬州大学学报:农业与生命科学版,2016,37(4):21-26.
[16]李晓锋,陈葳,李旭,等.宫颈癌细胞系中EMT相关基因的表达及其意义[J].西安交通大学学报(医学版),2017,38(2):210-214.
[17]Voutsadakis IA.Epithelial-mesenchymal transition(EMT)and regulation of EMT factors by steroid nuclear receptors in breast cancer:a review and in silico investigation[J].J Clin Med,2016,5(1):E11.
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[19]Chen CC,Sureshbabul M,Chen HW,et al.Curcumin suppresses metastasis via Sp-1,FAK inhibition,and E-Cadherin upregulation in colorectal cancer[J].Evid Based Complement Alternat Med,2013,2013:541695.
[20]陈劲松,黄炯强,董世濠,等.微小RNA-379-5p对肝癌细胞迁移和侵袭的影响[J].中华医学杂志,2016,96(18):1450-1453.
[21]张小博,李艳会,李开良,等.VEGF、MMP-2、MMP-9在胰腺癌中的表达及临床意义[J].中国老年学杂志,2014,34(2):327-329.
[2]李苗,刘芳芳,邸媛媛,等.DNA定量分析联合阴道镜在诊断宫颈癌及高级别宫颈上皮内瘤变中的价值[J].中国妇幼保健,2012,27(18):2841-2844.
[3]王翔宇,许晋铨,孙丽,等.Rsf-1/HBXAP基因在宫颈癌中的表达及临床病理意义[J].现代妇产科进展,2017,26(2):104-108.
[4]陈瑛,王丹丹,朱虹,等.抗肿瘤新靶点黏着斑激酶FAK及其抑制剂研究进展[J].中国现代应用药学,2016,33(2):255-260.
[5]Shanthi E,Krishna MH,Arunesh GM,et al.Focal adhesion kinase inhibitors in the treatment of metastatic cancer:a patent review[J].Expert Opin Ther Pat,2014,24(10):1077-1100.
[6]李吉友,贾栋.抑制FAK的表达对人胃癌细胞SGC-7901凋亡的影响[J].现代肿瘤医学,2017,25(1):34-37.
[7]王静雯,王思念,孙锁柱,等.CXCR7沉默对He La细胞侵袭和迁移能力的影响及其在宫颈癌中的表达[J].国际妇产科学杂志,2017,44(2):163-166;后插1页.
[8]Schaller MD,Borgman CA,Cobb BS,et al.pp125FAK a structurally distinctive protein-tyrosine kinase associated with focal adhesions[J].Proc Natl Acad Sci U S A,1992,89(11):5192-5196.
[9]高宁,张静,杜娟,等.沉默黏着斑激酶促进人胃癌SGC-7901细胞骨架蛋白解聚及细胞形态损伤[J].中国组织化学与细胞化学杂志,2017,26(1):7-12.
[10]Brami-Cherrier K,Gervasi N,Arsenieva D,et al.FAK dimerization controls its kinase-dependent functions at focal adhesions[J].EMBO J,2014,33(4):356-370.
[11]Nader GP,Ezratty EJ,Gundersen GG.FAK,talin and PIPKIγregulate endocytosed integrin activation to polarize focal adhesion assembly[J].Nat Cell Biol,2016,18(5):491-503.
[12]Liu L,Zong C,Li B,et al.The interaction betweenβ1 integrins and ERK1/2 in osteogenic differentiation of human mesenchymal stem cells under fluid shear stress modelled by a perfusion system[J].J Tissue Eng Regen Med,2014,8(2):85-96.
[13]O’Brien S,Golubovskaya VM,Conroy J,et al.FAK inhibition with small molecule inhibitor Y15 decreases viability,clonogenicity,and cell attachment in thyroid cancer cell lines and synergizes with targeted therapeutics[J].Oncotarget,2014,5(17):7945-7959.
[14]Lee BY,Timpson P,Horvath LG,et al.FAK signaling in human cancer as a target for therapeutics[J].Pharmacol Ther,2015,146:132-149.
[15]郝臻凤,王娟,周留林,等.FAK在宫颈癌中的表达及临床意义[J].扬州大学学报:农业与生命科学版,2016,37(4):21-26.
[16]李晓锋,陈葳,李旭,等.宫颈癌细胞系中EMT相关基因的表达及其意义[J].西安交通大学学报(医学版),2017,38(2):210-214.
[17]Voutsadakis IA.Epithelial-mesenchymal transition(EMT)and regulation of EMT factors by steroid nuclear receptors in breast cancer:a review and in silico investigation[J].J Clin Med,2016,5(1):E11.
[18]诸葛春凤,刘诗权,谭林,等.Sph K1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.
[19]Chen CC,Sureshbabul M,Chen HW,et al.Curcumin suppresses metastasis via Sp-1,FAK inhibition,and E-Cadherin upregulation in colorectal cancer[J].Evid Based Complement Alternat Med,2013,2013:541695.
[20]陈劲松,黄炯强,董世濠,等.微小RNA-379-5p对肝癌细胞迁移和侵袭的影响[J].中华医学杂志,2016,96(18):1450-1453.
[21]张小博,李艳会,李开良,等.VEGF、MMP-2、MMP-9在胰腺癌中的表达及临床意义[J].中国老年学杂志,2014,34(2):327-329.