ZLM-7 inhibits the occurrence and angiogenesis of breast cancer through miR-212-3p/Sp1/VEGFA signal axis.
ZLM-7 inhibits the occurrence and angiogenesis of breast cancer through miR-212-3p/Sp1/VEGFA signal axis.
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ZLM-7通过miR-212-3p/Sp1/VEGFA信号轴抑制乳腺癌的发生和血管生成
DOI：
10.1186/s10020-020-00239-2
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发表时间：
2020-11-13
期刊：
影响因子：
--
通讯作者：
Luo ZY
中科院分区：
文献类型：
--
作者：
Li X;Zou ZZ;Wen M;Xie YZ;Peng KJ;Luo T;Liu SY;Gu Q;Li JJ;Luo ZY
乳腺癌是一种常见的恶性肿瘤，预后差。血管生成与实体瘤的生长和进展有关，并与预后有关。ZLM-7、SP1、VEGFA和miR-212- 3 p与BC的血管生成和增殖有关，但具体机制尚不清楚。本研究旨在揭示乳腺癌血管生成的调控机制。用10 nM ZLM-7处理BC细胞系8小时。Western blot检测蛋白表达水平，qRT-PCR检测RNA表达水平。使用miR-212- 3 p模拟物或miR-212 - 3 p抑制剂、使用pcDNA3.1载体的Sp1过表达来进行miR-212 - 3 p的过表达或抑制。通过共培养BC细胞系和HUVEC细胞来分析血管生成。为了评估miR-212- 3 p和Sp1之间的调控关系，进行双荧光素酶测定。此外，利用ChIP技术分析了Sp1与VEGFA的直接相互作用。transwell法检测细胞迁移和侵袭能力，克隆形成法检测细胞增殖能力。在BC细胞建立的小鼠异种移植瘤模型中，我们还通过免疫组化检测了血管生成标志物CD 31。ZLM-7可上调miR-212- 3 p表达，抑制BC的侵袭、迁移、增殖和血管生成，而miR-212- 3 p抑制剂可拮抗这种作用。miR-212- 3 p与Sp1的结合序列显示，miR-212- 3 p在蛋白和mRNA水平均抑制Sp1的表达。Sp1可与VEGFA相互作用并促进其表达。miR-212- 3 p过表达抑制BC细胞的迁移、侵袭、增殖和血管生成，而Sp1过表达则表现出相反的作用，并可拮抗miR-212- 3 p过表达的这些作用。ZLM-7降低VEGFA表达，这通过与miR-212- 3 p抑制剂共转染来挽救。同样，ZLM-7在体内可通过miR-212- 3 p/Sp1/VEGFA轴抑制肿瘤生长和血管生成。ZLM-7可直接上调BC中miR-212- 3 p的表达。miR-212- 3 p通过Sp1抑制VEGFA表达，从而抑制BC的血管生成和进展。
Breast cancer (BC) is a common malignant tumor with poor prognosis. Angiogenesis is related to the growth and progression of solid tumors and associated with prognosis. ZLM-7, SP1, VEGFA and miR-212-3p were associated with BC angiogenesis and proliferation, however the detailed mechanism was not clear. This study aimed to reveal the regulatory mechanism of angiogenesis of BC. BC cell lines were treated with 10 nM ZLM-7 for 8 h. We detected protein expression level by western blot and RNA expression level by qRT-PCR. Overexpression or inhibition of miR-212-3p is performed using miR-212-3p mimics or miR-212-3p inhibitor, Sp1 overexpression using pcDNA3.1 vector. Angiogenesis was analyzed by co-culturing BC cell lines and HUVEC cells. To evaluate regulatory relationship between miR-212-3p and Sp1, dual luciferase assay was performed. Besides, the direct interaction between Sp1 and VEGFA was analyzed by ChIP. Migration and invasion were analyzed by transwell assay and proliferation was detected by clone formation assay. In mice xenograft model developed using BC cells, we also detected angiogenesis marker CD31 through immunohistochemistry. ZLM-7 up-regulated miR-212-3p and inhibited invasion, migration, proliferation and angiogenesis of BC, while miR-212-3p inhibitor antagonized such effects. Binding sequence was revealed between miR-212-3p and Sp1, and expression of Sp1 was inhibited by miR-212-3p on both protein and mRNA level. Sp1 could interact with VEGFA and promoted its expression. Overexpression of miR-212-3p inhibited migration, invasion, proliferation and angiogenesis of BC cell lines, while Sp1 overexpression showed the opposite effect and could antagonize these effects of miR-212-3p overexpression. ZLM-7 decreased VEGFA expression, which was rescued by co-transfection with miR-212-3p inhibitor. Similar, ZLM-7 could inhibit tumor growth and angiogenesis through the miR-212-3p/Sp1/VEGFA axis in vivo. ZLM-7 could directly up-regulate miR-212-3p in BC. MiR-212-3p could inhibit VEGFA expression through Sp1, thereby inhibiting angiogenesis and progression of BC.
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影响因子：
4
作者：
Li P;Xu T;Zhou X;Liao L;Pang G;Luo W;Han L;Zhang J;Luo X;Xie X;Zhu K
通讯作者：
Zhu K
影响因子：
9.8
作者：
Ramjiawan RR;Griffioen AW;Duda DG
通讯作者：
Duda DG
影响因子：
--
作者：
Kulshrestha A;Katara GK;Ibrahim SA;Patil R;Patil SA;Beaman KD
通讯作者：
Beaman KD
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影响因子：
8.8
作者：
Wang, Di;Zheng, Jian;Liu, Yunhui
通讯作者：
Liu, Yunhui
影响因子：
3.1
作者：
Elghoroury, Eman A.;ElDine, Hesham G.;Ibrahim, Mona H.
通讯作者：
Ibrahim, Mona H.