title: Auphen and dibutyryl cAMP suppress growth of hepatocellular carcinoma by regulating expression of aquaporins 3 and 9 in vivo.
abstract: OBJECTIVE
  To investigate whether the regulation of aquaporin 3 (AQP3) and AQP9 induced by Auphen and dibutyryl cAMP (dbcAMP) inhibits hepatic tumorigenesis.
  
  
  METHODS
  Expression of AQP3 and AQP9 was detected by Western blot, immunohistochemistry (IHC), and RT-PCR in HCC samples and paired non-cancerous liver tissue samples from 30 hepatocellular carcinoma (HCC) patients. A xenograft tumor model was used in vivo. Nine nude mice were divided into control, Auphen-treated, and dbcAMP-treated groups (n = 3 for each group). AQP3 and AQP9 protein expression after induction of xenograft tumors was detected by IHC and mRNA by RT-PCR analysis. The terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay and histological evaluation were used to detect apoptosis of tumor cells, and the concentration of serum α-fetoprotein (AFP) was measured using RT-PCR and an ELISA kit.
  
  
  RESULTS
  The volumes and weights of tumors decreased significantly in the Auphen- and dbcAMP-treated mice compared with the control mice (P < 0.01). The levels of AQP3 were significantly lower in the Auphen treatment group, and levels of AQP9 were significantly higher in thedbcAMP treatment mice than in the control mice (P < 0.01). The reduction of AQP3 by Auphen and increase of AQP9 by dbcAMP in nude mice suppressed tumor growth of HCC, which resulted in reduced AFP levels in serum and tissues, and apoptosis of tumor cells in the Auphen- and dbcAMP-treated mice, when compared with control mice (P < 0.01). Compared with para-carcinoma tissues, AQP3 expression increased in tumor tissues whereas the expression of AQP9 decreased. By correlating clinicopathological and expression levels, we demonstrated that the expression of AQP3 and AQP9 was correlated with clinical progression of HCC and disease outcomes.
  
  
  CONCLUSIONS
  AQP3 increases in HCC while AQP9 decreases. Regulation of AQP3 and AQP9 expression by Auphen and dbcAMP inhibits the development and growth of HCC.
authors.affiliation: Rui Peng, Guang-Xi Zhao, Jing Li, Yu Zhang, Xi-Zhong Shen, Ji-Yao Wang, Jian-Yong Sun, Department of Gastroenterology, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Peng
authors.affiliation: Rui Peng, Guang-Xi Zhao, Jing Li, Yu Zhang, Xi-Zhong Shen, Ji-Yao Wang, Jian-Yong Sun, Department of Gastroenterology, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
authors.forename: Guang-Xi
authors.initials: GX
authors.lastname: Zhao
authors.affiliation: Rui Peng, Guang-Xi Zhao, Jing Li, Yu Zhang, Xi-Zhong Shen, Ji-Yao Wang, Jian-Yong Sun, Department of Gastroenterology, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.affiliation: Rui Peng, Guang-Xi Zhao, Jing Li, Yu Zhang, Xi-Zhong Shen, Ji-Yao Wang, Jian-Yong Sun, Department of Gastroenterology, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Rui Peng, Guang-Xi Zhao, Jing Li, Yu Zhang, Xi-Zhong Shen, Ji-Yao Wang, Jian-Yong Sun, Department of Gastroenterology, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
authors.forename: Xi-Zhong
authors.initials: XZ
authors.lastname: Shen
authors.affiliation: Rui Peng, Guang-Xi Zhao, Jing Li, Yu Zhang, Xi-Zhong Shen, Ji-Yao Wang, Jian-Yong Sun, Department of Gastroenterology, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
authors.forename: Ji-Yao
authors.initials: JY
authors.lastname: Wang
authors.affiliation: Rui Peng, Guang-Xi Zhao, Jing Li, Yu Zhang, Xi-Zhong Shen, Ji-Yao Wang, Jian-Yong Sun, Department of Gastroenterology, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
authors.forename: Jian-Yong
authors.initials: JY
authors.lastname: Sun
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D051400:Aquaporin 3 / Q000235:genetics / Q000378:metabolism*; D020346:Aquaporins / Q000235:genetics / Q000378:metabolism*; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D003563:Cyclic CMP / Q000031:analogs & derivatives* / Q000494:pharmacology; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008113:Liver Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D008297:Male; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008875:Middle Aged; D050607:Organogold Compounds / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D047368:Tumor Burden / Q000187:drug effects; D023041:Xenograft Model Antitumor Assays; D000509:alpha-Fetoproteins / Q000378:metabolism
chemical_list: C097353:AFP protein, human; C496027:AQP3 protein, human; C109669:AQP9 protein, human; D000970:Antineoplastic Agents; D020346:Aquaporins; D050607:Organogold Compounds; D012333:RNA, Messenger; D000509:alpha-Fetoproteins; C000613556:dichloro(phenanthroline)gold(III); D051400:Aquaporin 3; D003563:Cyclic CMP; C053441:dibutyryl cyclic-3',5'-cytidine monophosphate
keywords: Aquaporin 3; Aquaporin 9; Auphen; Dibutyryl cAMP; Hepatocellular carcinoma; Nude mice; Xenograft tumor model