title: Genomic aberrations in the HTPAP promoter affect tumor metastasis and clinical prognosis of hepatocellular carcinoma.
abstract: We previously reported that the intronic tagSNP +357G/C in the metastasis suppressor HTPAP is associated with metastasis and prognosis of hepatocellular carcinoma (HCC). The aim of this study was to investigate whether SNPs in the HTPAP promoter modulate HTPAP expression and prognosis of HCC. Genomic DNA from 572 microdissected HCCs were genotyped by pyrosequencing and verified by direct sequencing. Haplotype blocks were analyzed. Reporter plasmids were constructed and transfected into HCC cell lines. Transcriptional activities of plasmids were analyzed by dual-luciferase reporter systems. HTPAP expression was measured by real-time quantitative PCR, western blots, and tissue microarrays. Invasion was assessed by Matrigel assays. The prognostic values of HTPAP promoter SNPs in HCC were evaluated by Kaplan-Meier and Cox regression analyses. We identified six SNPs, including -1053A/G and +64G/C, in the HTPAP promoter. The SNPs were in complete linkage disequilibrium, resulting in three promoter haplotypes (promoter I:-1053AA/+64GG, promoter II: -1053AG/+64GC, and promoter III: -1053GG/+64CC). Promoter I manifested the highest luciferase index (p<0.005). However, no significant difference was observed between promoters II and III. We consistently found that HTPAP mRNA and protein levels were significantly higher in promoter I than that of promoter II+III (p<0.001). Invasion was increased in HCC cells transfected with promoters II+III compared to those transfected with promoter I (p<0.05). The HTPAP promoter II+III haplotype was associated with significantly increased metastasis compared to that of promoter I (p = 0.023). The postoperative five-year overall survival of patients with promoters II+III was lower than that of patients with promoter I (p = 0.006). Multivariate analysis showed that the promoter II+III haplotype was an adverse prognostic marker in HCC. The genetic variants at loci -1053 and +64 of the HTPAP promoter affect the expression of HTPAP, which might be a novel determinant and target for HCC prognosis.
authors.affiliation: Department of Hepatobiliary Surgery, Hainan Provincial People's Hospital, Nanhua University, Haikou, People's Republic of China.
authors.forename: Jin-Cai
authors.initials: JC
authors.lastname: Wu
authors.affiliation: Liver Cancer Institute, Zhongshan Hospital, Fudan University, Key Laboratory of Carcinogenesis and Cancer Invasion (Fudan University), Ministry of Education, Shanghai, People's Republic of China; Cancer Center, Institute of Biomedical Sciences, Fudan University, Shanghai, People's Republic of China.
authors.forename: Hu-Liang
authors.initials: HL
authors.lastname: Jia
authors.affiliation: Department of Hepatobiliary Surgery, Hainan Provincial People's Hospital, Nanhua University, Haikou, People's Republic of China.
authors.forename: Zhuo-Ri
authors.initials: ZR
authors.lastname: Li
authors.affiliation: Department of Hepatobiliary Surgery, Hainan Provincial People's Hospital, Nanhua University, Haikou, People's Republic of China.
authors.forename: Kai-Lun
authors.initials: KL
authors.lastname: Zhou
authors.affiliation: Liver Cancer Institute, Zhongshan Hospital, Fudan University, Key Laboratory of Carcinogenesis and Cancer Invasion (Fudan University), Ministry of Education, Shanghai, People's Republic of China; Cancer Center, Institute of Biomedical Sciences, Fudan University, Shanghai, People's Republic of China.
authors.forename: Lun-Xiu
authors.initials: LX
authors.lastname: Qin
authors.affiliation: Liver Cancer Institute, Zhongshan Hospital, Fudan University, Key Laboratory of Carcinogenesis and Cancer Invasion (Fudan University), Ministry of Education, Shanghai, People's Republic of China; Cancer Center, Institute of Biomedical Sciences, Fudan University, Shanghai, People's Republic of China.
authors.forename: Qiong-Zhu
authors.initials: QZ
authors.lastname: Dong
authors.affiliation: Liver Cancer Institute, Zhongshan Hospital, Fudan University, Key Laboratory of Carcinogenesis and Cancer Invasion (Fudan University), Ministry of Education, Shanghai, People's Republic of China; Cancer Center, Institute of Biomedical Sciences, Fudan University, Shanghai, People's Republic of China.
authors.forename: Ning
authors.initials: N
authors.lastname: Ren
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D045744:Cell Line, Tumor; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D023281:Genomics; D006239:Haplotypes / Q000235:genetics; D006801:Humans; D008113:Liver Neoplasms / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D010711:Phosphatidate Phosphatase / Q000235:genetics* / Q000378:metabolism; D020641:Polymorphism, Single Nucleotide; D011379:Prognosis; D011401:Promoter Regions, Genetic / Q000235:genetics*; D014158:Transcription, Genetic / Q000235:genetics
chemical_list: C523991:PLPP5 protein, human; D010711:Phosphatidate Phosphatase