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

• 1:贵州医科大学附属医院肝胆外科贵州医科大学肝胆胰脾重点实验室贵州省肝胆胰脾疾病研究所

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

DOI: 10.19367/j.cnki.2096-8388.2020.08.002

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