Zhang Ying, Liu Yao, Guo Xingrong, Hu Zhenhua, Shi Huirong
Department of Gynaecology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450000, People's Republic of China.
Department of Gynaecology, Hami Central Hospital, Hami, Xinjiang 839000, People's Republic of China.
Onco Targets Ther. 2020 May 13;13:4145-4155. doi: 10.2147/OTT.S239979. eCollection 2020.
The dysregulation of the human papillomavirus 18 E6 and E7 oncogenes plays a critical role in the angiogenesis of cervical cancer (CC), including the proliferation, migration, and tube formation of vascular endothelial cells. Interfering E6/E7 increases the number of CC cell-derived microvesicles (CC-MVs). Additionally, microRNAs (miRNAs) can modulate CC angiogenesis and can be encapsulated in MVs.
We aim to investigate whether E6/E7 affects CC angiogenesis via regulating miRNAs in CC-MVs.
CC-MVs were isolated from a CC cell line (HeLa) which were transfected with small interfering RNAs (siRNAs) against E6/E7 or co-transfected with miR-377 mimics/inhibitors. The expression of several miRNAs in CC-MVs was detected using quantitative real-time PCR. After co-incubating CC-MVs with human umbilical vein endothelial cells (HUVECs), cell proliferation, migration, and tube formation of HUVECs were determined using cell counting kit-8, transwell, and tube formation assays, respectively.
MiR-377 was increased in E6/E7-interfering CC-MVs. Overexpressing miR-377 in CC-MVs suppressed HUVEC proliferation, migration, and tube formation. LPAR2, the cell surface G protein-coupled receptor, was the downstream target of miR-377 in HUVECs. The co-transfection of E6/E7 siRNAs and miR-377 inhibitors in CCs negated the effect of E6/E7 siRNAs on the elevation of miR-377 in CC-MVs. In HUVECs, the co-transfection of E6/E7 siRNAs and miR-377 inhibitors restored the LPAR2 expression which was reduced by the E6/E7 siRNA transfection. Meanwhile, miR-377 mimic reduced LPAR2 expression and inhibited HUVEC proliferation, migration, and tube formation, while such response was negated by LPAR2 overexpression.
Interfering E6/E7 increased miR-377 in CC-MVs, and overexpressing miR-377 in CC-MVs inhibited angiogenesis of HUVECs via reducing LPAR2.
人乳头瘤病毒18 E6和E7癌基因的失调在宫颈癌（CC）血管生成中起关键作用，包括血管内皮细胞的增殖、迁移和管腔形成。干扰E6/E7可增加CC细胞衍生微泡（CC-MVs）的数量。此外，微小RNA（miRNAs）可调节CC血管生成并可封装在微泡中。
我们旨在研究E6/E7是否通过调节CC-MVs中的miRNAs影响CC血管生成。
从CC细胞系（HeLa）中分离CC-MVs，该细胞系用针对E6/E7的小干扰RNA（siRNAs）转染或与miR-377模拟物/抑制剂共转染。使用定量实时PCR检测CC-MVs中几种miRNAs的表达。将CC-MVs与人脐静脉内皮细胞（HUVECs）共孵育后，分别使用细胞计数试剂盒-8、Transwell和管腔形成试验测定HUVECs的细胞增殖、迁移和管腔形成。
在干扰E6/E7的CC-MVs中miR-377增加。在CC-MVs中过表达miR-377可抑制HUVECs的增殖、迁移和管腔形成。LPAR2是细胞表面G蛋白偶联受体，是HUVECs中miR-377的下游靶点。在CC中，E6/E7 siRNAs与miR-377抑制剂共转染可消除E6/E7 siRNAs对CC-MVs中miR-377升高的影响。在HUVECs中，E6/E7 siRNAs与miR-377抑制剂共转染可恢复因E6/E7 siRNA转染而降低的LPAR2表达。同时，miR-377模拟物降低LPAR2表达并抑制HUVECs的增殖、迁移和管腔形成，而LPAR2过表达可消除这种反应。
干扰E6/E7可增加CC-MVs中的miR-377，在CC-MVs中过表达miR-377可通过降低LPAR2抑制HUVECs的血管生成。
