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长链非编码RNA AFAP1-AS1促进胆管癌细胞的生长和转移。

LncRNA AFAP1-AS1 promotes growth and metastasis of cholangiocarcinoma cells.

作者信息

Shi Xiuhui, Zhang Hang, Wang Min, Xu Xiaodong, Zhao Yan, He Ruizhi, Zhang Min, Zhou Min, Li Xu, Peng Feng, Shi Chengjian, Shen Ming, Wang Xin, Guo Xingjun, Qin Renyi

机构信息

Department of Biliary-Pancreatic Surgery, Affiliated Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.

出版信息

Oncotarget. 2017 Apr 6;8(35):58394-58404. doi: 10.18632/oncotarget.16880. eCollection 2017 Aug 29.

DOI:10.18632/oncotarget.16880
PMID:28938565
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5601661/
Abstract

We investigated the role of actin filament associated protein 1 antisense RNA1 (AFAP1-AS1) lncRNA in promoting cholangiocarcinoma (CCA). qRT-PCR analysis of patient samples showed that AFAP1-AS1 expression was higher in CCA tumors than matched adjacent non-tumor tissue. AFAP1-AS1 levels were also higher in CCA cell lines (HuCCT1 and TFK-1) than a normal biliary epithelium cell line (HIBEpic). AFAP1-AS1 knockdown in CCA cell lines using shAFAP1-AS1 reduced cell proliferation and colony formation in CCK-8 and colony formation assays, respectively. Cell cycle analysis demonstrated that AFAP1-AS1 knockdown resulted in G0/G1 cell cycle arrest and inhibition of S-G2/M transition compared to the controls. CCA cells transfected with shAFAP1-AS1 also exhibited reduced metastasis and invasiveness in Transwell and wound healing assays. This was further confirmed in xenograft experiments with nude mice using CCA cells transfected with shAFAP1-AS1 or control shRNA. AFAP1-AS1 knockdown cells produced smaller tumors, demonstrating that AFAP1-AS1 promotes tumor growth . AFAP1-AS1 knockdown also increased expression of actin filament associated protein 1 (AFAP1) and reduced cell stress filament integrity, as determined from western blot and immunofluorescence assays, respectively. These findings indicate that AFAP1-AS1 exerts oncogenic effects in CCA. We postulate that AFAP1-AS1 is a potentially useful diagnostic and prognostic biomarker and therapeutic target for CCA.

摘要

我们研究了肌动蛋白丝相关蛋白1反义RNA1（AFAP1-AS1）长链非编码RNA在促进胆管癌（CCA）发生中的作用。对患者样本进行的qRT-PCR分析显示，AFAP1-AS1在CCA肿瘤中的表达高于配对的癌旁非肿瘤组织。AFAP1-AS1在CCA细胞系（HuCCT1和TFK-1）中的水平也高于正常胆管上皮细胞系（HIBEpic）。在CCA细胞系中使用shAFAP1-AS1敲低AFAP1-AS1，分别在CCK-8和集落形成试验中降低了细胞增殖和集落形成。细胞周期分析表明，与对照组相比，AFAP1-AS1敲低导致G0/G1期细胞周期停滞并抑制S-G2/M期转换。在Transwell和伤口愈合试验中，用shAFAP1-AS1转染的CCA细胞也表现出转移和侵袭能力降低。在用shAFAP1-AS1或对照shRNA转染的CCA细胞对裸鼠进行的异种移植实验中，这一点得到了进一步证实。AFAP1-AS1敲低的细胞产生的肿瘤较小，表明AFAP1-AS1促进肿瘤生长。分别通过蛋白质印迹和免疫荧光分析确定，AFAP1-AS1敲低还增加了肌动蛋白丝相关蛋白1（AFAP1）的表达并降低了细胞应激丝的完整性。这些发现表明AFAP1-AS1在CCA中发挥致癌作用。我们推测AFAP1-AS1是一种潜在有用的CCA诊断和预后生物标志物及治疗靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ab6/5601661/edcb1c101f4b/oncotarget-08-58394-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ab6/5601661/aa41b3e25142/oncotarget-08-58394-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ab6/5601661/287c05fb38b7/oncotarget-08-58394-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ab6/5601661/05afe22724d2/oncotarget-08-58394-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ab6/5601661/a0344359960a/oncotarget-08-58394-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ab6/5601661/d97782a29aff/oncotarget-08-58394-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ab6/5601661/c4d623fcd0a4/oncotarget-08-58394-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ab6/5601661/edcb1c101f4b/oncotarget-08-58394-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ab6/5601661/aa41b3e25142/oncotarget-08-58394-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ab6/5601661/287c05fb38b7/oncotarget-08-58394-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ab6/5601661/05afe22724d2/oncotarget-08-58394-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ab6/5601661/a0344359960a/oncotarget-08-58394-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ab6/5601661/d97782a29aff/oncotarget-08-58394-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ab6/5601661/c4d623fcd0a4/oncotarget-08-58394-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ab6/5601661/edcb1c101f4b/oncotarget-08-58394-g007.jpg

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