王琴容,宋科伟,吴昌学,赵艳,谢渊,周建奖
WANG Qinrong,SONG Kewei,WU Changxue,ZHAO Yan,XIE Yuan,ZHOU Jianjiang

• 1:贵州医科大学地方病与少数民族性疾病教育部重点实验室
• 2:贵州医科大学贵州省医学分子生物学重点实验室
• 3:贵州医科大学运动与健康学院

摘要(Abstract)：

目的:探讨ALX1(aristaless-like homeobox 1)基因过表达对非小细胞肺癌细胞系PC-9上皮间质转化(EMT)能力的影响。方法:构建pc DNA3.1-ALX1过表达重组载体,将pc DNA3.1-ALX1和pc DNA3.1分别转染PC-9细胞;采用细胞划痕实验和Transwell实验分析PC-9细胞迁移及侵袭能力的改变,用qRT-PCR和Western印迹法检测EMT过程相关标志物N-钙黏蛋白(N-cadherin)、紧密连接蛋白(ZO-1)以及转录因子slug和snail表达水平的改变。结果:与对照组相比,ALX1过表达组中PC-9细胞的迁移和侵袭能力明显增强(P<0.001),ZO-1表达量明显降低(P<0.05),N-cadherin、slug和snail表达量明显增高(P<0.001)。结论:ALX1基因过表达能够促进非小细胞肺癌细胞PC-9的EMT转化,其机制可能与ZO-1表达量下调和N-cadherin、slug和snail表达量上调有关。
Objective: To explore the effects of ALX1( Aristaless-like homeobox 1) overexpression on the EMT capability of non-small cell lung cancer cell line PC-9. Methods: The ALX1 overexpression vector( pc DNA3. 1-ALX1) was constructed. Then pc DNA3. 1-ALX1 and pc DNA3. 1 were transfected into PC-9 cells using transfection reagent. The migration and invasion abilities of PC-9 cells were analyzed by wound healing assay and invasion assay,respectively. The expression changes of EMT hallmarks,including N-cadherin,ZO-1,slug and snail,were detected by qRT-PCR and Western blotting. Results: Compared with the control group,the migration and invasion abilities of PC-9 cells in ALX1 overexpression group significantly increased( P < 0. 001). The expression of ZO-1 was decreased but the expression of N-cadherin,slug and snail increased after ALX1 overexpression( P <0. 001). Conclusion: ALX1 overexpression could promote the EMT of PC-9 cells,and the mechanism may be related to the down-regulation of ZO-1 and up-regulation of N-cadherin,slug and snail.

关键词(KeyWords)： 癌,非小细胞肺;ALX1;上皮间质转化;侵袭;迁移
ALX1;epithelial-mesenchymal transformation;non-small cell lung cancer;invasion;migration

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(No.81660474);; 贵州省科技合作计划项目[黔科合LH字(2016)7347];; 贵州医科大学博士启动基金(院博合J字004)

作者(Author): 王琴容,宋科伟,吴昌学,赵艳,谢渊,周建奖
WANG Qinrong,SONG Kewei,WU Changxue,ZHAO Yan,XIE Yuan,ZHOU Jianjiang

DOI: 10.19367/j.cnki.1000-2707.2017.04.009

参考文献(References)：

• [1]Shtivelman E,Hensing T,Simon GR,et al.Molecular pathways and therapeutic targets in lung cancer[J].Oncotarget,2014(6):1392-1433.
• [2]Dela Cruz CS,Tanoue LT,Matthay RA.Lung cancer:epidemiology,etiology,and prevention[J].Clinics in chest medicine,2011(4):605-644.
• [3]Larsen JE,Minna JD.Molecular biology of lung cancer:clinical implications[J].Clinics in chest medicine,2011(4):703-740.
• [4]Ding W,You H,Dang H,et al.Epithelial-to-mesenchymal transition of murine liver tumor cells promotes invasion[J].Hepatology(Baltimore,Md),2010(3):945-953.
• [5]Li J,Wang Q,Wen R,et al.MiR-138 inhibits cell proliferation and reverses epithelial-mesenchymal transition in non-small cell lung cancer cells by targeting GIT1and SEMA4C[J].Journal of cellular and molecular medicine,2015(12):2793-2805.
• [6]Yang M,Pan Y,Zhou Y.Depletion of ALX1 causes inhibition of migration and induction of apoptosis in human osteosarcoma[J].Tumour biology,2015(8):5965-5970.
• [7]Yuan H,Kajiyama H,Ito S,et al.ALX1 induces snail expression to promote epithelial-to-mesenchymal transition and invasion of ovarian cancer cells[J].Cancer research,2013(5):1581-1590.
• [8]管停宏,苏琳茜,易永芬,等.软骨同源蛋白1(CART1)敲低诱导MDA-MB-231乳腺癌细胞S期阻滞并抑制其侵袭和迁移[J].细胞与分子免疫学杂志,2016(4):474-478.
• [9]Sandoval J,Mendez-Gonzalez J,Nadal E,et al.A prognostic DNA methylation signature for stage I nonsmall-cell lung cancer[J].Journal of clinical oncology,2013(32):4140-4147.
• [10]Cai RL.Human CART1,a paired-class homeodomain protein,activates transcription through palindromic binding sites[J].Biochemical and biophysical research communications,1998(2):305-311.
• [11]Regnier CH,Tomasetto C,Moog-Lutz C,et al.Presence of a new conserved domain in CART1,a novel member of the tumor necrosis factor receptor-associated protein family,which is expressed in breast carcinoma[J].The Journal of biological chemistry,1995(43):25715-25721.
• [12]Uz E,Alanay Y,Aktas D,et al.Disruption of ALX1causes extreme microphthalmia and severe facial clefting:expanding the spectrum of autosomal-recessive ALXrelated frontonasal dysplasia[J].American journal of human genetics,2010(5):789-796.
• [13]Zhao GQ,Zhou X,Eberspaecher H,et al.Cartilage homeoprotein 1,a homeoprotein selectively expressed in chondrocytes[J].Proceedings of the National Academy of Sciences of the United States of America,1993(18):8633-8637.
• [14]Zhao Q,Behringer RR,de Crombrugghe B.Prenatal folic acid treatment suppresses acrania and meroanencephaly in mice mutant for the Cart1 homeobox gene[J].Nature genetics,1996(3):275-283.

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