Long non-coding RNA UBE2CP3 enhances HCC cell secretion of VEGFA and promotes angiogenesis by activating ERK1/2/HIF-1α/VEGFA signalling in hepatocellular carcinoma.
Long non-coding RNA UBE2CP3 enhances HCC cell secretion of VEGFA and promotes angiogenesis by activating ERK1/2/HIF-1α/VEGFA signalling in hepatocellular carcinoma.
复制标题
长非编码RNA UBE2CP3通过激活肝细胞癌中的ERK1/2/HIF-1 α/VEGFA信号增强HCC细胞分泌VEGFA并促进血管生成
DOI：
10.1186/s13046-018-0727-1
复制
发表时间：
2018-06-04
期刊：
影响因子：
--
通讯作者：
Zheng L
中科院分区：
文献类型：
--
作者：
Lin J;Cao S;Wang Y;Hu Y;Liu H;Li J;Chen J;Li P;Liu J;Wang Q;Zheng L
血管生成被认为是恶性肿瘤发展的重要过程，与肿瘤的进展和转移有关。肝细胞癌（Hepatocellular carcinoma，HCC）是肝脏最常见的原发性肿瘤，是一种典型的血管生成性肿瘤。因此，研究肝癌血管生成的机制具有重要意义。长链非编码RNA（lncRNA）泛素结合酶E2 C假基因3（UBE 2CP 3）已被报道为促进HCC中的肿瘤转移的癌基因。然而，UBE 2CP 3在HCC血管生成中的作用和潜在机制仍不清楚。我们通过原位杂交（ISH）和实时定量聚合酶链反应（qRT-PCR）检测HCC患者样本中UBE 2CP 3的表达水平。我们还同时使用CD 31/PAS双染色来测量内皮血管（EV）密度，并使用qRT-PCR来测量CD 31 mRNA水平。用Lv-UBE 2CP 3或Sh-UBE 2CP 3病毒转染HepG 2和SMMC-7721细胞以获得稳定过表达或敲低的UBE 2CP 3细胞系。采用0.4 μm孔径的Transwell小室建立共培养体系，研究UBE 2CP 3对内皮细胞的间接作用。共培养体系中的HCC细胞和EC是分离的，但细胞因子和生长因子能够相互交流。暴露于肝癌细胞后，收集EC进行功能研究。最后，我们通过鸡胚绒毛尿囊膜（CAM）血管生成实验和裸鼠致瘤实验研究了UBE 2CP 3在体内的功能。在这项研究中，我们发现UBE 2CP 3在HCC组织中的表达高于癌旁组织，并且在EV密度高的组织中上调。在功能上，我们发现在共培养系统中，过表达UBE 2CP 3的HCC细胞通过激活ERK/HIF-1α/p70 S6 K/VEGFA信号通路促进HUVEC增殖、迁移和管腔形成，增加HCC细胞上清液中VEGFA的水平。此外，当UBE 2CP 3在HCC细胞中的表达被敲低时，出现相反的结果。与这些结果一致，CAM血管生成测定和裸鼠致瘤性测定显示UBE 2CP 3表达上调体内EV密度。我们的研究表明UBE 2CP 3可以增强HCC肿瘤细胞和HUVEC之间的相互作用，并通过促进血管生成来促进HCC的致瘤性。本文的在线版本（10.1186/s13046-018-0727-1）包含补充材料，可供授权用户使用。
Angiogenesis is considered as an important process in the development of malignancies and is associated with cancer progression and metastasis. Hepatocellular carcinoma (HCC) is the most common primary tumor of the liver and is recognized as a typical angiogenic tumor. Thus, it is of great importance to study the underlying mechanism of angiogenesis in HCC. The long non-coding RNA (lncRNA) ubiquitin conjugating enzyme E2C pseudogene 3 (UBE2CP3) has been reported as an oncogene that promotes tumor metastasis in HCC. However, the role and underlying mechanisms of UBE2CP3 in HCC angiogenesis are still unclear. We measured the expression levels of UBE2CP3 by in situ hybridization (ISH) and quantitative real-time polymerase chain reaction (qRT-PCR) in HCC patient samples. We also concomitantly used CD31/PAS double-staining to measure endothelial vessel (EV) density and used qRT-PCR to measure the CD31 mRNA level. HepG2 and SMMC-7721 cells were transfected with Lv-UBE2CP3 or Sh-UBE2CP3 virus to obtain stably over-expressing or knocking-down UBE2CP3 cell lines. The indirect effects of UBE2CP3 on ECs were studied by establishing a co-culture system using Transwell chambers with a 0.4-μm pore size. HCC cells and ECs in the co-culture system were separated, but the cytokines and growth factors were able to communicate with each other. Following exposed to HCC cells, ECs were collected for functional studies. Finally, we studied the function of UBE2CP3 in vivo by chick embryo chorioallantoic membrane (CAM) angiogenesis assays and nude mouse tumorigenicity assays. In this study, we found that UBE2CP3 expression was higher in HCC tissues than in para-tumor tissues and was up-regulated in tissues with high EV density. Functionally, we found that in the co-culture systems, HCC cells overexpressing UBE2CP3 promoted HUVEC proliferation, migration and tube formation via the activation of ERK/HIF-1α/p70S6K/VEGFA signalling, increasing the level of VEGFA in HCC cell supernatant. In addition, the opposite results appeared when the expression of UBE2CP3 in HCC cells was knocked down. Consistent with these results, CAM angiogenesis assays and nude mouse tumorigenicity assays showed that UBE2CP3 expression up-regulated EV density in vivo. Our study suggests that UBE2CP3 can enhance the interaction between HCC tumor cells and HUVECs and promote HCC tumorigenicity by facilitating angiogenesis. The online version of this article (10.1186/s13046-018-0727-1) contains supplementary material, which is available to authorized users.
登录
查看更多内容
影响因子：
--
作者：
Cao SW;Huang JL;Chen J;Hu YW;Hu XM;Ren TY;Zheng SH;Lin JD;Tang J;Zheng L;Wang Q
通讯作者：
Wang Q
影响因子：
8
作者：
通讯作者：
--
影响因子：
37.3
作者：
Hoshino Y;Hayashida T;Hirata A;Takahashi H;Chiba N;Ohmura M;Wakui M;Jinno H;Hasegawa H;Maheswaran S;Suematsu M;Kitagawa Y
通讯作者：
Kitagawa Y
前列腺癌中脂质运载蛋白 2 的过度表达通过激活 ERK 信号传导增加 SLUG 表达来促进细胞迁移和侵袭
影响因子：
2.8
作者：
Ding, Guanxiong;Fang, Jie;Liu, Jun
通讯作者：
Liu, Jun
DOI：
10.1007/978-981-10-1498-7_8
发表时间：
2016-01-01
期刊：
LONG AND SHORT NONCODING RNAS IN CANCER BIOLOGY
影响因子：
--
作者：
Khorshidi, Azam;Dhaliwal, Preet;Yang, Burton B.
通讯作者：
Yang, Burton B.