title: The long non-coding RNA HOTTIP promotes progression and gemcitabine resistance by regulating HOXA13 in pancreatic cancer.
abstract: BACKGROUND
  The human genome encodes many long non-coding RNAs (lncRNAs). However, their biological functions, molecular mechanisms, and the prognostic value associated with pancreatic ductal adenocarcinoma (PDAC) remain to be elucidated. Here, we identify a fundamental role for the lncRNA HOXA transcript at the distal tip (HOTTIP) in the progression and chemoresistance of PDAC.
  
  
  METHODS
  High-throughput microarrays were performed to detect the expression profiles of lncRNAs and messenger RNAs in eight human PDAC tissues and four pancreatic tissues. Quantitative real-time PCR was used to determine the levels of HOTTIP and HOXA13 transcripts in PDAC cell lines and 90 PDAC samples from patients. HPDE6 cells (immortalized human pancreatic ductal epithelial cells) and corresponding adjacent non-neoplastic tissues were used as controls, respectively. The functions of HOTTIP and HOXA13 in cell proliferation, invasion, and epithelial-mesenchymal transition were evaluated by targeted knockdown in vitro. CCK-8 assays, colony formation assays, and xenografts in nude mice were used to investigate whether targeted silencing of HOTTIP could sensitize pancreatic cancer cells to gemcitabine. Immunohistochemistry was performed to investigate the relationship between HOXA13 expression and patient outcome.
  
  
  RESULTS
  Microarray analyses revealed that HOTTIP was one of the most significantly upregulated lncRNAs in PDAC tissues compared with pancreatic tissues. Quantitative PCR further verified that HOTTIP levels were increased in PDAC cell lines and patient samples compared with controls. Functionally, HOTTIP silencing resulted in proliferation arrest by altering cell-cycle progression, and impaired cell invasion by inhibiting epithelial-mesenchymal transition in pancreatic cancer. Additionally, inhibition of HOTTIP potentiated the antitumor effects of gemcitabine in vitro and in vivo. Furthermore, knockdown of HOXA13 by RNA interference (siHOXA13) revealed that HOTTIP promoted PDAC cell proliferation, invasion, and chemoresistance, at least partly through regulating HOXA13. Immunohistochemistry results revealed that higher HOXA13 expression was correlated with lymph node metastasis, poor histological differentiation, and decreased overall survival in PDAC patients.
  
  
  CONCLUSIONS
  As a crucial tumor promoter, HOTTIP promotes cell proliferation, invasion, and chemoresistance by modulating HOXA13. Therefore, the HOTTIP/HOXA13 axis is a potential therapeutic target and molecular biomarker for PDAC.
authors.affiliation: Department of Medical Oncology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China. lizhihuasysu@126.com.
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Li
authors.affiliation: Department of Medical Oncology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China. zhxiaohui_sysu@163.com.
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Zhao
authors.affiliation: Department of Pancreaticobiliary Surgery, Hepatobiliary Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China. zyzysmile@163.com.
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Department of Radiotherapy, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China. Lyimin_sysu@126.com.
authors.forename: Yimin
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Pancreaticobiliary Surgery, Hepatobiliary Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China. zhquanbo@126.com.
authors.forename: Quanbo
authors.initials: Q
authors.lastname: Zhou
authors.affiliation: Department of Pancreaticobiliary Surgery, Hepatobiliary Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China. Yhuilin_sysu@126.com.
authors.forename: Huilin
authors.initials: H
authors.lastname: Ye
authors.affiliation: Department of Pancreaticobiliary Surgery, Hepatobiliary Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China. wyingxue_sysu@126.com.
authors.forename: YinXue
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Medical Oncology, Zengcheng People's Hospital, Sun Yat-sen University, Guangzhou, China. zjinglong_sysu@126.com.
authors.forename: Jinlong
authors.initials: J
authors.lastname: Zeng
authors.affiliation: Department of Medical Oncology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China. syadong_sysu@126.com.
authors.forename: Yadong
authors.initials: Y
authors.lastname: Song
authors.affiliation: Department of Pancreaticobiliary Surgery, Hepatobiliary Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China. gwenchao_sysu@126.com.
authors.forename: Wenchao
authors.initials: W
authors.lastname: Gao
authors.affiliation: Department of Pancreaticobiliary Surgery, Hepatobiliary Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China. zhshangyou_sysu@126.com.
authors.forename: ShangYou
authors.initials: S
authors.lastname: Zheng
authors.affiliation: Department of Pancreaticobiliary Surgery, Hepatobiliary Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China. 18820095121@163.com.
authors.forename: Baoxiong
authors.initials: B
authors.lastname: Zhuang
authors.affiliation: Department of Medical Oncology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China. chhuimou_sysu@126.com.
authors.forename: Huimou
authors.initials: H
authors.lastname: Chen
authors.affiliation: Department of Medical Oncology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China. lwenzhu_sysu@126.com.
authors.forename: Wenzhu
authors.initials: W
authors.lastname: Li
authors.affiliation: Department of Pathology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China. lhaigang_sysu@126.com.
authors.forename: Haigang
authors.initials: H
authors.lastname: Li
authors.affiliation: Department of Pathology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China. lhaifeng_sysu@126.com.
authors.forename: Haifeng
authors.initials: H
authors.lastname: Li
authors.affiliation: Department of Pancreaticobiliary Surgery, Hepatobiliary Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China. 496763865@qq.com.
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Fu
authors.affiliation: Department of Pancreaticobiliary Surgery, Hepatobiliary Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China. chenrf_sysu@163.com.|Department of General Surgery, The Second Affiliated Hospital of Sun Yat-sen University, Sun Yat-sen University, 107 Yan-Jiang Xi Road, Guangzhou, 510120, China. chenrf_sysu@163.com.
authors.forename: Rufu
authors.initials: R
authors.lastname: Chen
mesh_terms: D000818:Animals; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D018450:Disease Progression; D019008:Drug Resistance, Neoplasm / Q000187:drug effects*; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects; D005260:Female; D055785:Gene Knockdown Techniques; D018398:Homeodomain Proteins / Q000378:metabolism*; D006801:Humans; D008297:Male; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008875:Middle Aged; D009361:Neoplasm Invasiveness; D010190:Pancreatic Neoplasms / Q000473:pathology*; D062085:RNA, Long Noncoding / Q000378:metabolism*; D016019:Survival Analysis; D000093542:Gemcitabine
chemical_list: D018398:Homeodomain Proteins; D062085:RNA, Long Noncoding; C099959:homeobox protein HOXA13; D003841:Deoxycytidine; D000093542:Gemcitabine