LncRNA AFAP1-AS1 promotes growth and metastasis of cholangiocarcinoma cells
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Xiuhui Shi1,*, Hang Zhang1,*, Min Wang1,*, Xiaodong Xu1, Yan Zhao1, Ruizhi He1, Min Zhang1, Min Zhou1, Xu Li1, Feng Peng1, Chengjian Shi1, Ming Shen1, Xin Wang1, Xingjun Guo1 and Renyi Qin1
1Department of Biliary-Pancreatic Surgery, Affiliated Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
*These authors have contributed equally to this study
Renyi Qin, email: firstname.lastname@example.org
Xingjun Guo, email: email@example.com
Keywords: long noncoding RNA, AFAP1-AS1, cholangiocarcinoma, proliferation, metastasis
Received: February 07, 2017 Accepted: March 15, 2017 Published: April 06, 2017
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 in vivo. 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.
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