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Minoru Matsubara,
Hidenori Shiraha,
Jyunro Kataoka,
Masaya Iwamuro,
Shigeru Horiguchi,
Shin-Ichi Nishina,
Nobuyuki Takaoka, Masayuki Uemura,
Akinobu Takaki,
Shinichiro Nakamura,
Yoshiyuki Kobayashi,
Kazuhiro Nouso,
Kazuhide Yamamoto
[show abstract]
[hide abstract]
ABSTRACT: Background and Aim: Hepatocellular carcinoma (HCC) is a hypervascular tumor, and angiogenesis plays an important role in its development. Previously, we demonstrated that des-γ-carboxyl prothrombin (DCP) promotes both cell proliferation and migration of human umbilical vein endothelial cells (HUVECs) by inducing the autophosphorylation of kinase insert domain receptor (KDR). In the present study, DCP-associated tumor angiogenesis was assessed by comparing hypovascular and common hypervascular HCC. Methods: The solitary HCCs of 827 patients were classified into two groups according to the tumor density at the arterial phase of a dynamic computed tomography scan; the initial clinical data of patients with the hyper- and hypovascular types were compared. The HCC tissues from 95 tumors were analyzed by immunohistochemical staining for DCP and phosphorylated KDR, and intratumoral microvessel density (MVD) was analyzed to evaluate microvessel angiogenesis. Results: The serum DCP levels (320 ± 3532 mAU/mL) and tumor size (18.4 ± 9.0 mm) of patients with hypervascular HCC were significantly greater than those with hypovascular HCC (38.7 ± 80 mAU/mL and 14.6 ± 5.2 mm, P < 0.001). Immunohistochemical analysis revealed that the expressions of DCP and phospho-KDR were significantly greater in hypervascular HCC (71.4% and 31.0%, respectively) than in hypovascular HCC (7.6% and 5.7%, respectively). Intratumoral MVD was significantly correlated with DCP (r = 0.48, P < 0.0001). Conclusions: des-γ-carboxyl prothrombin production is associated with tumor angiogenesis in HCC.
Journal of Gastroenterology and Hepatology 05/2012; 27(10):1602-8. · 2.87 Impact Factor
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Shin-Ichi Nishina,
Hidenori Shiraha,
Yutaka Nakanishi,
Shigetomi Tanaka,
Minoru Matsubara,
Nobuyuki Takaoka, Masayuki Uemura,
Shigeru Horiguchi,
Junro Kataoka,
Masaya Iwamuro,
Takahito Yagi,
Kazuhide Yamamoto
[show abstract]
[hide abstract]
ABSTRACT: Runt-related transcription factor 3 (RUNX3) is a candidate tumor suppressor gene that is downregulated in various cancers. In the present study, we analyzed the regulatory function of RUNX3 on Jagged-1 (JAG1) expression and cancer stem cell (CSC) signaling in hepatocellular carcinoma (HCC). Eleven HCC cell lines and 30 human HCC tissues were used. RUNX3 and JAG1 expression levels were analyzed by immunoblotting and immunohistochemistry. Ectopic RUNX3 expression was induced by introducing RUNX3 cDNA into the RUNX3-negative HCC cell line Hep3B and Huh7 cells. Furthermore endogenous RUNX3 expression was knocked down by RUNX3 siRNA in SK-Hep-1 cells. In order to analyze JAG1 transcriptional regulation, we conducted reporter assays, chromatin immunoprecipitation (ChIP) assays and electrophoretic mobility shift assays (EMSAs). Tumorigenicity was analyzed using a SCID mouse liver injection model. An inverse correlation was observed between RUNX3 expression and JAG1 expression in most HCC cell lines and tissues. Restoring RUNX3 expression decreased the expression of JAG1 in Hep3B and Huh7 cells, whereas JAG1 expression was upregulated in RUNX3 siRNA-treated SK-Hep-1 cells. Reporter assays, ChIP assays and EMSAs revealed that RUNX3 directly bound to the transcriptional regulatory region of JAG1 and suppressed JAG1 transcription. Moreover, RUNX3 restoration downregulated CSCs by suppressing JAG1-mediated Notch signaling. The tumorigenic capacity of RUNX3-expressing Hep3B cells was lower compared to that of control Hep3B cells. RUNX3 expression suppressed JAG1 expression and resulted in downregulation of tumorigenesis by suppression of JAG1-mediated CSCs.
Oncology Reports 05/2011; 26(3):523-31. · 1.84 Impact Factor
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Yutaka Nakanishi,
Hidenori Shiraha,
Shin-ichi Nishina,
Shigetomi Tanaka,
Minoru Matsubara,
Shigeru Horiguchi,
Masaya Iwamuro,
Nobuyuki Takaoka, Masayuki Uemura,
Kenji Kuwaki,
Hiroaki Hagihara,
Junichi Toshimori,
Hideki Ohnishi,
Akinobu Takaki,
Shinichiro Nakamura,
Yoshiyuki Kobayashi,
Kazuhiro Nouso,
Takahito Yagi,
Kazuhide Yamamoto
[show abstract]
[hide abstract]
ABSTRACT: Runt-related transcription factor 3 (RUNX3) is known as a tumor suppressor gene for gastric cancer and other cancers, this gene may be involved in the development of hepatocellular carcinoma (HCC).
RUNX3 expression was analyzed by immunoblot and immunohistochemistry in HCC cells and tissues, respectively. Hep3B cells, lacking endogenous RUNX3, were introduced with RUNX3 constructs. Cell proliferation was measured using the MTT assay and apoptosis was evaluated using DAPI staining. Apoptosis signaling was assessed by immunoblot analysis.
RUNX3 protein expression was frequently inactivated in the HCC cell lines (91%) and tissues (90%). RUNX3 expression inhibited 90±8% of cell growth at 72 h in serum starved Hep3B cells. Forty-eight hour serum starvation-induced apoptosis and the percentage of apoptotic cells reached 31±4% and 4±1% in RUNX3-expressing Hep3B and control cells, respectively. Apoptotic activity was increased by Bim expression and caspase-3 and caspase-9 activation.
RUNX3 expression enhanced serum starvation-induced apoptosis in HCC cell lines. RUNX3 is deleted or weakly expressed in HCC, which leads to tumorigenesis by escaping apoptosis.
BMC Cancer 01/2011; 11:3. · 3.01 Impact Factor
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Ryuta Takenaka,
Yoshiro Kawahara,
Hiroyuki Okada,
Keisuke Hori,
Masafumi Inoue,
Seiji Kawano,
Daisuke Tanioka,
Takao Tsuzuki, Masayuki Uemura,
Nobuya Ohara,
Susumu Tominaga,
Tomoo Onoda,
Kazuhide Yamamoto
[show abstract]
[hide abstract]
ABSTRACT: The narrow-band imaging (NBI) system is a novel technology that enhances the visualization of microvasculature and mucosal patterns. The aim of this study was to assess the reliability of the NBI system for esophageal cancer screening in patients with head and neck cancers.
A total of 142 patients with head and neck squamous cell carcinoma (SCC) were examined by NBI endoscopy, followed by Lugol chromoendoscopy between April 2006 and June 2008 at the Okayama University Hospital, Okayama, Japan. Detection of SCC and high-grade intraepithelial neoplasia (HGIN) was conducted.
The median age of the patients was 64 years (range: 29-86 years), and approximately three-fourths of all the patients were male. In total, 21 superficial lesions in 16 patients were detected by NBI endoscopy. Of these, 4 lesions were diagnosed histologically as SCC and 11 lesions as HGIN. An additional 22 Lugol-voiding lesions >or=5 mm were detected in 19 patients by Lugol chromoendoscopy. Although 1 of these lesions was diagnosed as HGIN, 21 lesions were diagnosed as low-grade intraepithelial neoplasia or lesions without atypical findings. The sensitivity of NBI endoscopy for detecting esophageal SCC and HGIN was 90.9% (95% confidence interval (CI), 58.7-99.8), specificity was 95.4% (95% CI, 90.3-98.3), and accuracy was 95.1% (95% CI, 90.1-98.0).
NBI seems to be useful and reliable for screening for esophageal SCC in patients with head and neck cancers.
The American Journal of Gastroenterology 08/2009; 104(12):2942-8. · 7.28 Impact Factor
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Ryuta Takenaka,
Yoshiro Kawahara,
Hiroyuki Okada,
Keisuke Hori,
Masafumi Inoue,
Seiji Kawano,
Daisuke Tanioka,
Takao Tsuzuki, Masayuki Uemura,
Nobuya Ohara,
Susumu Tominaga,
Tomoo Onoda,
Kazuhide Yamamoto
[show abstract]
[hide abstract]
ABSTRACT: OBJECTIVES: The narrow-band imaging (NBI) system is a novel technology that enhances the visualization of microvasculature and mucosal patterns. The aim of this study was to assess the reliability of the NBI system for esophageal cancer screening in patients with head and neck cancers.
The American Journal of Gastroenterology 07/2009; 104(12):2942-2948. · 7.28 Impact Factor
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Noriyuki Matsuo,
Hidenori Shiraha,
Tatsuya Fujikawa,
Nobuyuki Takaoka,
Naoki Ueda,
Shigetomi Tanaka,
Shinichi Nishina,
Yutaka Nakanishi, Masayuki Uemura,
Akinobu Takaki,
Shinichiro Nakamura,
Yoshiyuki Kobayashi,
Kazuhiro Nouso,
Takahito Yagi,
Kazuhide Yamamoto
[show abstract]
[hide abstract]
ABSTRACT: Twist, a transcription factor of the basic helix-loop-helix class, is reported to regulate cancer metastasis. It is known to induce epithelial-mesenchymal transition (EMT). In this study, we evaluated the expression of twist and its effect on cell migration in hepatocellular carcinoma (HCC).
We examined twist expression using immunohistochemistry in 20 tissue samples of hepatocellular carcinoma, and assessed twist expression in HCC cell lines by RT-PCR and Western blot analysis. Ectopic twist expression was created by introducing a twist construct in the twist-negative HCC cell lines. Endogenous twist expression was blocked by twist siRNA in the twist-positive HCC cell lines. We studied EMT related markers, E-cadherin, Vimentin, and N-cadherin by Western blot analysis. Cell proliferation was measured by MTT assay, and cell migration was measured by in vitro wound healing assay. We used immunofluorescent vinculin staining to visualize focal adhesion.
We detected strong and intermediate twist expression in 7 of 20 tumor samples, and no significant twist expression was found in the tumor-free resection margins. In addition, we detected twist expression in HLE, HLF, and SK-Hep1 cells, but not in PLC/RPF/5, HepG2, and Huh7 cells. Ectopic twist-expressing cells demonstrated enhanced cell motility, but twist expression did not affect cell proliferation. Twist expression induced epithelial-mesenchymal transition together with related morphologic changes. Focal adhesion contact was reduced significantly in ectopic twist-expressing cells. Twist-siRNA-treated HLE, HLF, and SK-Hep1 cells demonstrated a reduction in cell migration by 50, 40 and 18%, respectively.
Twist induces migratory effect on hepatocellular carcinoma by causing epithelial-mesenchymal transition.
BMC Cancer 02/2009; 9:240. · 3.01 Impact Factor
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Naoki Ueda,
Hidenori Shiraha,
Tatsuya Fujikawa,
Nobuyuki Takaoka,
Yutaka Nakanishi,
Mayumi Suzuki,
Noriyuki Matsuo,
Shigetomi Tanaka,
Shin-Ichi Nishina, Masayuki Uemura,
Akinobu Takaki,
Yasushi Shiratori,
Kazuhide Yamamoto
[show abstract]
[hide abstract]
ABSTRACT: Using GGCX gene-specific real-time PCR, exon 2 deletion splice variant of vitamin K-dependent gamma-glutamyl carboxylase (GGCX) mRNA was identified in HCC cell lines. Expressions of wild type and exon 2 deletion variant of GGCX were analyzed with relevance to DCP production in HCC cell lines. Hep3B, HepG2, HuH1, HuH7, and PLC/PRF/5 produced DCP, while SK-Hep-1, HLE, HLF, and JHH1 produced no detectable level of DCP. DCP-producing cells expressed exon 2 deletion variant of GGCX mRNA and protein, while DCP-negative cells expressed no detectable level of exon 2 deletion variant of GGCX. These results suggest that exon 2 deletion splice variant of GGCX causes dysfunction of GGCX enzyme activity resulting in DCP production in HCC cell lines.
Molecular oncology 11/2008; 2(3):241-9. · 4.10 Impact Factor
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Ryuta Takenaka,
Yoshiro Kawahara,
Hiroyuki Okada,
Keisuke Hori,
Masafumi Inoue,
Seiji Kawano,
Daisuke Tanioka,
Takao Tsuzuki,
Satoru Yagi,
Jun Kato, Masayuki Uemura,
Nobuya Ohara,
Tadashi Yoshino,
Atsushi Imagawa,
Shigeatsu Fujiki,
Rie Takata,
Kazuhide Yamamoto
[show abstract]
[hide abstract]
ABSTRACT: Although endoscopic submucosal dissection (ESD) is expected to reduce the local recurrence of gastric cancers, we still experience cases of recurrence after an ESD.
To characterize clinical and pathologic features of cases with local recurrence of early gastric cancer after an ESD.
A prospective cohort study.
A total of 306 patients with gastric cancers removed by ESD at Okayama University Hospital and Tsuyama Central Hospital between March 2001 and December 2005 were enrolled.
ESD.
Local recurrence.
The incidence of a complete en bloc resection was 80.4% when pathologically evaluated. Within a median follow-up period of 26 months (12-64 months), a local recurrence was found in 7 cases, all of which had been declared incomplete resections. One patient underwent a second ESD, and the remaining 6 underwent a surgical resection. All removed lesions were mucosal cancers. No lymph-node metastases were found in patients with a surgical resection. There was a significant correlation between the incidence of an incomplete resection and that of a local recurrence (P < .0001). Among the clinical characteristics, tumor size (>30 mm vs <20 mm; odds ratio [OR] 16 mm [95% CI, 2.0-130 mm]) and tumor location (upper vs middle or lower; OR 7.6 [95% CI, 1.3-45]) were identified as factors that were significantly associated with the incidence of a local recurrence.
Short follow-up duration.
The incidence of a local recurrence was strongly associated with that of an incomplete resection. The frequency of a local recurrence also showed significant correlations with the tumor size and location within the stomach.
Gastrointestinal endoscopy 06/2008; 68(5):887-94. · 6.71 Impact Factor
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Eiichiro Yumoto,
Harushige Nakatsukasa,
Tadashi Hanafusa,
Yasuhiro Yumoto,
Kazuhiro Nouso,
Eiji Matsumoto,
Toru Onishi,
Yoshitaka Takuma,
Hironori Tanaka,
Tatsuya Fujikawa,
Mayumi Suzuki, Masayuki Uemura,
Yasushi Shiratori
[show abstract]
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ABSTRACT: Insulin-like growth factor binding protein-3 (IGFBP-3) is a mediator of growth suppression signals and a putative tumor suppressor gene. The growth suppression mechanisms of IGFBP-3 have not been well clarified. We examined the expression of IGFBP-3 transcripts in human hepatocellular carcinoma (HCC) and the relationship between IGFBP-3 expression and the transforming growth factor-beta (TGF-beta) and/or retinoblastoma (Rb) signaling pathways. In situ hybridization revealed IGFBP-3 transcripts in cancer cells in 6 of 57 (10%) HCCs, including moderately and poorly differentiated HCCs with intrahepatic metastasis. In contrast, all lung metastatic nodules of 4 HCCs showed IGFBP-3 transcripts in cancer cells. The cDNA microarray showed that genes for the TGF-beta pathway and Rb were up-regulated in IGFBP-3-expressing HCCs. In 6 HCCs presenting IGFBP-3, immunohistochemical analyses showed abnormalities in the TGF-beta and/or Rb pathways; the loss of phosphorylated-Smad2 was observed in 2, and overexpression of phosphorylated-Rb was observed in the remaining 4 HCCs. The present study suggests that IGFBP-3 mediates growth suppression signals via the TGF-beta and/or Rb pathways in HCC.
International Journal of Oncology 12/2005; 27(5):1223-30. · 2.40 Impact Factor
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Keishi Fujiwara,
Toshihiro Higashi,
Kazuhiro Nouso,
Harushige Nakatsukasa,
Yoshiyuki Kobayashi, Masayuki Uemura,
Shin-Ichiro Nakamura,
Shuichiro Sato,
Tadashi Hanafusa,
Yasuhiro Yumoto,
Ichiro Naito,
Yasushi Shiratori
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ABSTRACT: We analyzed the expression of antigen-processing and antigen-presenting molecules in surgically resected fresh samples of human hepatocellular carcinoma (HCC) tissue to elucidate a mechanism of immune escape. We also examined the expression of interleukin (IL)-10 protein, which might act to downregulate expression of antigen-processing and antigen-presenting molecules.
Twenty-eight HCC samples obtained by surgical resection were analyzed for the expression of beta2-microglobulin, heat-shock protein (HSP)-70, human leukocyte antigen (HLA) class-I, CD80 (B7-1), CD86 (B7-2) and IL-10 by immunostaining.
Beta2-microglobulin and HSP-70 were preserved in all samples. In contrast, the expression of HLA class-I molecules was significantly reduced according to lowering in the histological grading of tumor differentiation (P = 0.024). Furthermore, B7-1 and B7-2 expression was reduced in tumor cells compared with corresponding areas of liver tissue without malignant involvement irrespective of the histological grading of tumors (21% and 36%, respectively). Although IL-10 protein was expressed in 54% of HCC, no relationship between the expression of IL-10 and downregulation of B7-1, B7-2, and HLA class-I was evident.
These findings suggest the potential role of B7 co-stimulatory molecules and HLA class-I molecules in facilitating HCC escape from immune surveillance without the involvement of IL-10.
Journal of Gastroenterology and Hepatology 11/2004; 19(10):1121-7. · 2.87 Impact Factor
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Eiji Matsumoto,
Harushige Nakatsukasa,
Kazuhiro Nouso,
Shin-Ichiro Nakamura,
Mayumi Suzuki,
Yoshiyuki Kobayashi, Masayuki Uemura,
Syuichiro Sato,
Ei-Ichiro Yumoto,
Junko Yokoyama,
Sou Tsuboi,
Hironori Tanaka,
Yoshiataka Takuma,
Tatsuya Fujikawa,
Yasushi Shiratori
Comparative Hepatology 02/2004; 3 Suppl 1:S51. · 1.88 Impact Factor
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Toru Onishi,
Kazuhiro Nouso,
Toshihiro Higashi,
Nobuyuki Toshikuni,
Harushige Nakatsukasa,
Yoshiyuki Kobayashi, Masayuki Uemura,
Eiichiro Yumoto,
Keishi Fujiwara,
Shuichiro Sato,
Shinichiro Nakamura,
Junko Yokoyama,
Tadashi Hanafusa,
Yasushi Shiratori
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ABSTRACT: Telomerase is the enzyme that synthesizes telomeric DNA, and the activation of telomerase is closely related to cellular immortality. Telomerase activity has been reported in many human cancer tissues and is regulated by the expression of human telomerase reverse transcriptase (hTERT). The aim of the present study was to identify hTERT-expressing cells in human liver tissues and evaluate the feasibility of the hTERT promoter for gene therapy of hepatocellular carcinoma (HCC).
The authors examined the cellular distribution of hTERT transcripts in surgically resected HCC by in situ hybridization.
Among 20 samples, hTERT expression was observed in 15 HCC. Transcripts of hTERT were homogenously distributed in the cytoplasm of HCC cells in nine of 15 cases; six of 15 cases displayed a heterogeneous staining pattern. All poorly differentiated HCC that expressed hTERT showed a homogenous pattern of staining. None of the non-cancerous hepatocytes were positive for the transcripts, but infiltrating lymphocytes were faintly stained. The homogenous expression of hTERT was also observed in the vascular invasion of HCC.
The results indicate that most HCC cells express hTERT RNA and that the promoter is a good candidate as a target for gene therapy. However, careful consideration must be taken concerning the potential effects on lymphocytes.
Journal of Gastroenterology and Hepatology 11/2003; 18(10):1168-74. · 2.87 Impact Factor
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[show abstract]
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ABSTRACT: To identify antigens specifically recognized by the immune surveillance system in patients with hepatocellular carcinoma (HCC), the authors examined two complementary DNA (cDNA) libraries of moderately differentiated HCC by serologic analysis of recombinant cDNA expression libraries (SEREX).
The libraries were screened with autologous patients' sera, and sequences of the reacted clones were determined. To study the immunoreactivity of the antigens, sera from 20 patients with HCC, from 20 healthy volunteers, and from 16 patients with chronic viral hepatitis were examined.
Twenty-seven antigens were identified. They included SART1, p57Kip2, ROCK-1, gamma-catenin, and heat shock proteins, which are classified as tumor-associated genes. Three of 27 antigens-Tat-binding protein-1 (TBP-1), beta4 integrin-binding protein (p27[BBP]), and ribosomal protein L30 (rpL30)-were reacted predominantly with sera from patients with HCC (55% of patients, 45% of patients, and 20% of patients, respectively). Patients in the control group had no antibodies against these three antigens. Seventy percent of patients with HCC had the antibody against at least one of these antigens.
Disease specific humoral immune response against TBP-1, p27(BBP), and rpL30 was induced in patients with HCC, and the antibodies against these antigens also may be used as tumor markers.
Cancer 06/2003; 97(10):2474-9. · 4.77 Impact Factor
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Tadashi Hanafusa,
Yasuhiro Yumoto,
Kazuhiro Nouso,
Harushige Nakatsukasa,
Toru Onishi,
Tatsuya Fujikawa,
Mayumi Taniyama,
Shinichiro Nakamura, Masayuki Uemura,
Yoshitaka Takuma,
Eiichiro Yumoto,
Toshihiro Higashi,
Takao Tsuji
[show abstract]
[hide abstract]
ABSTRACT: Insulin-like growth factor binding protein-3 (IGFBP-3) is postulated to be a mediator of growth suppression signals. Reduced expression of the IGFBP-3 was observed in nine out of 12 human hepatocellular carcinomas (HCC) (75%). Promoter hypermethylation of the IGFBP-3 was detected in four out of 12 HCCs (33%) although mutations were not identified. The expression of IGFBP-3 was restored by the demethylating agent 5-aza-2'-deoxycytidine in HCC cell line with promoter hypermethylation (HepG2). As IGFBP-3 functions like a tumor suppressor gene, it may be used as a therapeutic target for HCC.
Cancer Letters 03/2002; 176(2):149-58. · 4.24 Impact Factor
