贵州医科大学学报

2020, v.45;No.239(08) 875-881

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敲低LEF1-AS1对胰腺癌细胞的影响
The Effect of Knocking down LEF1-AS1 on Pancreatic Cancer Cells and its Mechanism

杨哲豪;喻超;潘耀振;邓路;郑迪杰;孙诚谊;
YANG Zhehao;YU Chao;PAN Yaozhen;DENG Lu;ZHEN Dijie;SUN Chenyi;Department of Hepatobiliary Surgery,the Affiliated Hospital of Guizhou Medical University Key Laboratory of Liver,Gallbladder,Pancreas and Spleen,Guizhou Medical University Institute of Liver,Gallbladder,Pancreas and Spleen Diseases Diseases of Guizhou;

摘要(Abstract):

目的:探究淋巴增强因子1反义RNA1(LEF1-AS1)对胰腺癌(PC)细胞的影响及机制。方法:使用Gene Expression Profiling Interactive Analysis(GEPIA)网站分析来源于癌症基因组图谱(TCGA)数据库中PC组织和正常胰腺组织LEF1-AS1的表达;取对数生长期的正常人胰管上皮细胞株HPDE和PC细胞株Panc-1、Bxpc-3、As PC-1、Capan-1、CFPAC-1及MIA Pa Ca-2,采用q PCR检测LEF1-AS1在各细胞中的表达;选取q PCR检测后LEF1-AS1表达较高的PC细胞株Panc-1和MIA Pa Ca-2,用si LEF1-AS1与si Control分别转染后分为下调组和对照组,分别采用细胞计数试剂盒(CCK-8)、平板克隆实验、划痕实验和Transwell实验检测敲低LEF1-AS1对各组PC细胞增殖、细胞侵袭及迁移能力的影响,采用Western blot实验检测敲低LEF1-AS1对各组PC细胞中B细胞白血病淋巴瘤2(Bcl-2)、B细胞白血病淋巴瘤相关蛋白(Bax)、钙黏附蛋白E(E-cadherin)及波形蛋白(Vimentin)表达的影响。结果:PC组织中LEF1-AS1的表达高于正常组织(P <0.05),Panc-1、Bxpc-3、As PC-1、Capan-1、CFPAC-1及MIA Pa Ca-2细胞中LEF1-AS1表达水平较HPDE细胞上调(P <0.05);下调组Panc-1和MIA Pa Ca-2细胞24 h、48 h和72 h时OD值均较对照组下降,下调组细胞集落形成数较对照组降低(P <0.05);下调组Panc-1和MIA PaCa-2细胞迁移和侵袭能力较对照组细胞降低(P <0.05);敲低LEF1-AS1表达后,下调组Panc-1和MIA Pa Ca-2细胞Bcl-2和Vimentin蛋白的表达量均较与对照组下降,Bax和E-cadherin蛋白的表达量均较与对照组升高(P <0.05)。结论:敲低LEF1-AS1表达后抑制PC细胞的增殖、迁移和侵袭,其机制可能与促进PC细胞的凋亡和抑制上皮细胞-间充质转化进程有关。
Objective: To explore the effect and mechanism of lymphaden enhancer-binding factor 1 antisense RNA 1( LEF1-AS1) on pancreatic cancer( PC) cells. Methods: The expression of LEF1-AS1 in PC tissues and normal tissues based on the cancer genome atlas( TCGA) database was analyzed and compared by Gene Expression Profiling Interactive Analysis( GEPIA). The expression of LEF1-AS1 in PC cell lines Panc-1,Bxpc-3,As PC-1,Capan-1,CFPAC-1,MIA PaCa-2 and normal human pancreatic duct epithelial cell line HPDE was detected by qPCR. PC cell lines with high expression of LEF1-AS1 Panc-1 and MIA PaCa-2 were divided into the down-regulated group and the control group.The two groups were transfected by si LEF1-AS1 and si Control separately. The effects of LEF1-AS1 on proliferation,migration and invasion of PC cells of each group were detected by CCK-8 assay,plate clone formation assay,Transwell assay and scratch assay. The expression of Bcl-2,Bax,E-cadherin,Vimentin in PC cells of each group was detected by Western blot. Results: The expression of LEF1-AS1 in PC tissues was higher than that in normal pancreatic tissues( P < 0. 05). The expression of LEF1-AS1 in PC cell lines was higher than that in normal pancreatic epithelial cells( P < 0. 05). The OD values of down-regulated Panc-1 and MIA PaCa-2 cells in 24 h,48 h and 72 h were lower than those of the control group. The number of cell colony formation in the down-regulated group was lower than that in the control group( P < 0. 05). The migration and invasion of down-regulated Panc-1 and MIA PaCa-2 cells were significantly lower than those in the control group( P < 0. 05),and the expression of Bcl-2 and Vimentin proteins in down-regulated Panc-1 and MIA PaCa-2 cells was lower than that in the control group. The expression of Bax and E-cadherin proteins in down-regulated Panc-1 and MIA PaCa-2 cells was higher than that in the control group( P < 0. 05). Conclusion: Knocking down expression of LEF1-AS1 may inhibit the proliferation,migration and invasion of PC cells. The mechanism may be related to the promotion of apoptosis and the inhibition of the epithelialmesenchymal transformation process.

关键词(KeyWords): 胰腺肿瘤;细胞增殖;细胞迁移分析;LEF1-AS1基因;长链非编码RNA;侵袭
pancreatic neoplasms;cell proliferation;cell migration assays;LEF1-AS1 gene;long non-coding RNA(lncRNA);invasion

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(81860505,81860506);; 贵州省科学技术厅-贵州医科大学附属医院联合基金[黔科合LH字(2016)7229];贵州省肝胆外科临床医学研究中心[黔科合平台人才(2017)5404]

作者(Author): 杨哲豪;喻超;潘耀振;邓路;郑迪杰;孙诚谊;
YANG Zhehao;YU Chao;PAN Yaozhen;DENG Lu;ZHEN Dijie;SUN Chenyi;Department of Hepatobiliary Surgery,the Affiliated Hospital of Guizhou Medical University Key Laboratory of Liver,Gallbladder,Pancreas and Spleen,Guizhou Medical University Institute of Liver,Gallbladder,Pancreas and Spleen Diseases Diseases of Guizhou;

Email:

DOI: 10.19367/j.cnki.2096-8388.2020.08.002

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