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核因子白细胞介素3调节因子促进乳腺癌细胞增殖和侵袭能力研究

Nuclear factor interleukin 3-regulated promotes the proliferation and metastasis of breast cancer cells
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摘要 目的探讨核因子白细胞介素3调节因子(NFIL3)对人乳腺癌细胞增殖和侵袭能力的影响。方法采用慢病毒载体构建过表达及敲减NFIL3乳腺癌细胞系BT549和Hs578T(中国科学院上海细胞库),以荧光定量聚合酶链反应(PCR)及蛋白印迹实验(Western blot)进行验证。通过细胞计数试剂盒(MTT)检测细胞增殖、小室细胞迁移实验(Transwell)和侵袭实验以及动物模型裸鼠皮下荷瘤和鼠尾静脉肺转移实验,验证NFIL3对乳腺癌细胞增殖和侵袭能力。组间比较采用t检验及One-way ANOVA检验。结果MTT细胞增殖实验证实,过表达NFIL3组与对照组相比促进细胞增殖(BT549-对照4.37±0.27比BT549-NFIL35.98±0.31,t=6.783,P<0.01;Hs578T-对照4.75±0.32比Hs578T-NFIL36.05±0.39,t=4.463,P<0.05),敲减NFIL3抑制细胞增殖(BT549-PLKO 4.17±0.20比BT549-sh12.83±0.29,t=6.557,P<0.01;BT549-PLKO 4.17±0.20比BT549-sh23.11±0.17,t=4.943,P<0.01;Hs578T-PLKO 4.11±0.31比Hs578T-sh13.11±0.21,t=12.540,P<0.01;Hs578T-PLKO 4.11±0.31比Hs578T-sh23.01±0.23,t=10.930,P<0.01);Transwell细胞迁移证实,过表达NFIL3组与对照组比较促进细胞迁移[BT549-对照(198.00±28.03)个比BT549-NFIL3(397.00±52.10)个,t=10.640,P<0.01;Hs578T-对照(294.00±32.01)个比Hs578T-NFIL3(598.00±46.11)个,t=17.130,P<0.01],敲减NFIL3抑制细胞迁移[BT549-PLKO(178.00±23.11)个比BT549-sh1(41.01±15.12)个,t=9.820,P<0.01;BT549-PLKO(178.00±23.11)个比BT549-sh2(56.10±24.27)个,t=10.430,P<0.01;Hs578T-PLKO(197.60±21.13)个比Hs578T-sh1(69.80±19.96)个,t=8.403,P<0.01;Hs578T-PLKO(197.60±21.13)个比Hs578T-sh2(58.00±14.26)个,t=8.042,P<0.01];Transwell细胞侵袭证实,过表达NFIL3组与对照组比较促进细胞侵袭[BT549-对照(74.6±23.14)个比BT549-NFIL3(142.50±28.30)个,t=5.874,P<0.01;Hs578T-对照(103.20±28.14)个比Hs578T-NFIL3(183.70±22.14)个,t=7.110,P<0.01],敲减NFIL3抑制细胞侵袭[BT549-PLKO(89.13±19.86)个比BT549-sh1(37.45±15.63)个,t=9.062,P<0.01;BT549-PLKO(89.13±19.86)个比BT549-sh2(50.17±17.12)个 Objective To investigate the effects of Nuclear factor interleukin 3regulator(NFIL3)on proliferation and invasion of human breast cancer cells.Methods we successfully established cell models of NFIL3 over-expression and knockdown in breast cancer cell lines BT549 and Hs578T respectively(Shanghai cell bank of Chinese Academy of Sciences)thought lentiviral vectorsand validated by fluorescence quantitative polymerase chain reaction and Western blotting.Methyl thiazolyl tetrazolium assay(MTT)test cell proliferation,Transwell assays,Subcutaneous tumor bearing and tail vein lung metastasis in nude mice,were verified the proliferation and invasion of NFIL3 in breast cancer cells.Data between sample groups expressed by mean±standard deviation,and comparison between groups performed by T-test and One-way ANOVA test.Results MTT cell proliferation assay demonstrated that NFIL3 over-expression significantly promoted cell proliferation(BT549-control 4.37±0.27 vs.BT549-NFIL35.98±0.31,t=6.783,P<0.01;Hs578T-control 4.75±0.32 vs.Hs578T-NFIL36.05±0.39,t=4.463,P<0.05),and NFIL3 knockdown significantly inhibited cell proliferation(BT549-PLKO 4.17±0.20 vs.BT549-sh12.83±0.29,t=6.557,P<0.01;BT549-PLKO 4.17±0.20 vs.BT549-sh23.11±0.17,t=4.943,P<0.01;Hs578T-PLKO 4.11±0.31 vs.Hs578T-sh13.11±0.21,t=12.540,P<0.01;Hs578T-PLKO 4.11±0.31 vs.Hs578T-sh23.01±0.23,t=10.930,P<0.01).Transwell assay showed that NFIL3 over-expression significantly promoted cell migration(BT549-control 198.00±28.03 vs.BT549-NFIL3397.0±52.1,t=10.640,P<0.01;Hs578T-control 294±32.01 vs.Hs578T-NFIL3598±46.11,t=17.130,P<0.01),and oNFIL3 knockdown significantly inhibited cell migration(BT549-PLKO 178±23.11 vs.BT549-sh141.01±15.12,t=9.820,P<0.01;BT549-PLKO 178±23.11 vs.BT549-sh256.1±24.27,t=10.430,P<0.01;Hs578T-PLKO 197.6±21.13 vs.Hs578T-sh169.8±19.96,t=8.403,P<0.01;Hs578T-PLKO 197.6±21.13 vs.Hs578T-sh258.0±14.26,t=8.042,P<0.01).Transwell assay showed that NFIL3 over-expression significantly promoted cell invasion(BT549-control 74.6±23.14 vs.BT549-NFI
作者 宁宁 张雁凯 颜艺超 黄贞华 Ning Ning;Zhang Yankai;Yan Yichao;Huang Christine(Department of Gastroentestinal,Peking University International Hospital,Beijing 102206,China)
出处 《中华实验外科杂志》 CAS 北大核心 2022年第4期617-620,共4页 Chinese Journal of Experimental Surgery
基金 国家自然科学基金青年项目(81201688) 国家自然科学基金青年项目(81702336) 北京大学国际医院院内科研基金(YN2019XQ01) 北京大学国际医院院内课题中青年启动资助项目(YN2020QN08)。
关键词 乳腺癌 增殖 侵袭 Breast cancer Proliferation Metastasis
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