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  • 1
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 74, No. 19_Supplement ( 2014-10-01), p. 3612-3612
    Abstract: Background: Cancer-associated fibroblasts (CAF) have been suggested to be associated with the scirrhous gastric cancer (SGC) progression. SGC have a heterogeneously hypoxic environment which has been currently considered to be associated with aggressive tumor phenotypes cancer. Stromal cell-derived factor (SDF)-1 and C-X-C chemokine receptor type 4 (CXCR4) form an important chemokine/receptor pair, and are supposed to be up-regulated by hypoxia inducible factor-1 (HIF-1). The aim of our study was to clarify the effect of hypoxic environment on the CAF and CXCR4/SDF-1 axis in SGC. Experimental Design: Proliferation of SGC cells grown in monoculture and co-culture with CAF under hypoxia. SGC and CAF expression level of CXCR4 and SDF-1 under hypoxia were examined by RT-PCR and ELISA. Proliferation and migration of SGC cells with SDF-1 or hepatocyte growth factor (HGF) or conditioned medium from CAF under hypoxia, furthermore, added c-Met inhibitor or CXCR4 inhibitor. Results: The proliferation that co-culture of SGC cells with CAF was significantly increased under hypoxia. The expression level of CXCR4 mRNA was significantly increased under hypoxia in all of SGC cells. SDF-1 concentration level was significantly increased under hypoxia. The proliferation of SGC cells with SDF-1 and CAF was significantly increased. The proliferation of SGC cells with CAF was inhibited by CXCR4 inhibitor under hypoxia. The migration of SGC cells with HGF and CAF was significantly increased. The migration of SGC cells with CAF was inhibited by c-Met inhibitor. Conclusion: CAF might up-regulate the proliferation and migration of SGC cells under hypoxia, in comparison with that under normoxia. CXCR4/SDF-1 axis might play an important role for the proliferation of SGC cells under hypoxia. Citation Format: Haruhito Kinoshita, Masakazu Yashiro, Go Masuda, Hiroaki Kasashima, Tamami Morisaki, Tatsunari Fukuoka, Masatune Shibutani, Sadaaki Yamazoe, Katsunobu Sakurai, Hisashi Nagahara, Kenjiro Kimura, Takahiro Toyokawa, Ryosuke Amano, Naoshi Kubo, Hiroaki Tanaka, Kazuya Muguruma, Hiroshi Otani, Kiyoshi Maeda, Masaichi Ohira, Kosei Hirakawa. Effect of cancer associated fibroblast on gastric cancer cells under hypoxia. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3612. doi:10.1158/1538-7445.AM2014-3612
    Type of Medium: Online Resource
    ISSN: 0008-5472 , 1538-7445
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2014
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  • 2
    In: British Journal of Cancer, Springer Science and Business Media LLC, Vol. 113, No. 7 ( 2015-9), p. 1046-1054
    Type of Medium: Online Resource
    ISSN: 0007-0920 , 1532-1827
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    Language: English
    Publisher: Springer Science and Business Media LLC
    Publication Date: 2015
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  • 3
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 74, No. 19_Supplement ( 2014-10-01), p. 3606-3606
    Abstract: Introduction: Prostaglandins and cyclooxygenase2 have been suggested to play an important role for the proliferation and migration of cancer cells. But Prostaglandins (PG) have been suggested to affect cancer cells in various directions. For example, PGF2α and PGD2 has been demonstrated to have anti-tumor effects against some types of cancer cells. But PGE2 has been demonstrated to promote the proliferation and migration of cancer. Therefore, the investigation of the involvement of PG signals for cancer in lower level of arachidonic acid cascade is very important. The aim of this study is to investigate the effects of PGE2 and PGE2 receptor inhibitor (EP2 inhibitor) on the growth-interaction between cancer cells and fibroblasts. Methods: Three human gastric cancer cell lines (OCUM-2M, OCUM-2MD3, OCUM-12) and a human gastric fibroblast (CAF45) cell line were used in this study. Serum-free conditioned medium (SF-CM) from CAF-45 stimulated with PGE2 or EP2 inhibitor was prepared and used in study. Cell proliferation was studied by MTT assay and cell count in the presence or absence of CAF45. Cell migration was studied by wound healing assay in the presense or absence of SF-CM. In vivo study, We examined the effect of EP2 inhibitor on subcutaneous and orthotopically transplanted tumors. Results: In proliferation, PGE2 significantly stimulated the proliferation and migration of cancer cells. PGE2 significantly stimulated the proliferation and migration of cancer cells co-cultured with CAF45 or SF-CM more in comparison with single culture. EP2 inhibitor significantly decreased the proliferation and migration of cancer cells, EP2 inhibitor significantly decreased the proliferation migration of cancer cells co-cultured with CAF-45 or SF-CM more in comparison with single culture. In vivo study, EP2 inhibitor significantly suppressed the subcutaneous tumor in nude mice and suppressed orthotopic tumor growth and lymph node metastasis. Conclusion: PGE2 is associated with progression of gastric cancer. PGE2 receptor inhibitors might be promising anti-cancer drug for gastric carcinoma. Citation Format: Tatsunari Fukuoka, Masakazu Yashiro, Hiroshi Takeda, Takayuki Maruyama, Hiroaki Kasashima, Go Masuda, Haruhito Kinoshita, Tamami Morisaki, Katsunobu Sakurai, Masatsune Shibutani, Sadaaki Yamazoe, Kenjiro Kimura, Hisashi Nagahara, Takahiro Toyokawa, Ryosuke Amano, Naoshi Kubo, Hiroaki Tanaka, Kazuya Muguruma, Hiroshi Otani, Kiyoshi Maeda, Masaichi Ohira, Kosei Hirakawa. The effect of PGE2 receptor inhibitor on the microenviroment of scirrhous gastric cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3606. doi:10.1158/1538-7445.AM2014-3606
    Type of Medium: Online Resource
    ISSN: 0008-5472 , 1538-7445
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2014
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  • 4
    In: Journal of Clinical Oncology, American Society of Clinical Oncology (ASCO), Vol. 35, No. 4_suppl ( 2017-02-01), p. 74-74
    Abstract: 74 Background: The adjuvant chemotherapy with S-1 is the standard treatment for Stage II/III gastric cancer in Japan. Immunological status of host is critical for treatment outcome. Several investigators showed that systemic immune-inflammaotry indexes including neutrophil lymphocyte ratio (NLR) and modified Glasgow Prognostic Score (mGPS) well reflected the tumor progression. Methods: We analyzed clinical data obtained from 170 patients with pathological stage II/III gastric cancer who underwent surgery followed by S-1 adjuvant chemotherapy at Osaka City University Hospital between 2006 and 2015. Tumor infiltrating cells were detected by immunohistochemistry. Results: We found recurrent diseases in 70 (41%) patients including 15 in stage II and 55 in stage III. In univariate analysis using Cox proportion model, 2 grade of mGPS, the increase value of post-operative CEA, CA19-9, number of lymphocytes and NLR were associated with recurrence. Post-operative elevation of CEA and NLR were identified as independent risk factors for recurrence in multivariate analysis. Increase value of pre-operative NLR and CEA was significantly associated with early recurrent within one year after surgery. Tumor infiltrating neutrophils and PD-1+ T cells had correlated with the increase of pre-operative NLR and CEA value, respectively. Patients with low PD-1+T cells and low neutrophils had better prognosis than those with high infiltration. Conclusions: Post-surgical elevation of CEA and NLR value were useful as a predictive marker for recurrence in patients treated with S-1 adjuvant chemotherapy after surgery for gastric cancer. Early recurrence had correlated with tumor infiltrating neutrophils and PD-1+T cells. Our results suggested that systemic and local immune suppression should be an important element to exacerbate prognosis after chemotherapy for resectable gastric cancer.
    Type of Medium: Online Resource
    ISSN: 0732-183X , 1527-7755
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    Language: English
    Publisher: American Society of Clinical Oncology (ASCO)
    Publication Date: 2017
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  • 5
    In: Surgical Laparoscopy, Endoscopy & Percutaneous Techniques, Ovid Technologies (Wolters Kluwer Health), Vol. 24, No. 2 ( 2014-04), p. 158-163
    Type of Medium: Online Resource
    ISSN: 1530-4515
    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2014
    detail.hit.zdb_id: 2045171-4
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  • 6
    In: Surgical Laparoscopy, Endoscopy & Percutaneous Techniques, Ovid Technologies (Wolters Kluwer Health), Vol. 18, No. 3 ( 2008-06), p. 310-314
    Type of Medium: Online Resource
    ISSN: 1530-4515
    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2008
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  • 7
    In: Cancer Science, Wiley, Vol. 107, No. 8 ( 2016-08), p. 1101-1109
    Abstract: Tumor lymphangiogenesis is a major prognostic indicator of gastric cancer. Tumor‐induced inflammation has been shown to attract tumor‐associated macrophages that affect lymphangiogenesis. However, detailed mechanisms of macrophage‐induced lymphangiogenesis have not been elucidated. Here, we evaluated the interaction between tumor‐associated macrophages and lymphatic endothelial cells ( LEC s) derived from lymph nodes ( LN s) of human gastric cancer. Lymphatic endothelial cells were directly or indirectly cocultured with macrophages from healthy human blood, with or without the supernatant of the gastric cancer cell line, OCUM ‐12. We analyzed the effect of cancer pretreated macrophages and of macrophages from metastatic LN s of gastric cancer on LEC s. We observed morphological changes of LEC s in coculture and assessed the gene expression of possible lymphangiogenic molecules of macrophages and LEC s after contact coculture, and of cancer pretreated macrophages, by quantitative RT ‐ PCR . Specimens of metastatic LN of gastric cancer were immunofluorescently stained. We found that tubulogenesis of LEC s was observed only in the contact coculture model. OCUM ‐12 cells promoted macrophage‐induced tubulogenesis of LEC s. Relative gene expression of MMP and adhesion molecules was significantly upregulated in both capillary‐forming LEC s and cocultured macrophages. Cancer pretreated macrophages upregulated lymphangiogenic factors including inflammatory cytokines, MMP s, adhesion molecules, and vascular endothelial growth factor‐C. Blocking of intercellular adhesion molecule‐1 and macrophage activation suppressed tubulogenesis of LEC s. Immunohistochemistry showed macrophages localized around lymphatic vessels. Our results suggested that interaction between LEC s and macrophages may be an important initial step of tumor lymphangiogenesis developing LN metastasis. Understanding of its mechanisms could be useful for future therapeutics of gastric cancer.
    Type of Medium: Online Resource
    ISSN: 1347-9032 , 1349-7006
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2016
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  • 8
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 74, No. 19_Supplement ( 2014-10-01), p. 3924-3924
    Abstract: Background: Anaplastic pancreatic cancer is a rare malignancy with a poor prognosis. As a whole, pancreatic cancer is a devastating prognosis but its therapeutic strategy has not still well developed. Thus, new models are needed to research its biology. Purpose: We have established and characterized 2 anaplastic cancer cell lines (OCUP-A1 and OCUP-A2) for revealing tumor worsening factors and the mechanism of dedifferentiation. Methods: We researched the present studies by comparing with established 2 cell lines and well-known pancreatic cancer cell lines. In vitro proliferation, migration and invasion assays was examined in each cell lines under normoxia and hypoxia by CellPlayer™ 96-Well Kinetic Cell Proliferation, Migration and Invasion Assays. Furthermore,as assessment of chemosensitivity, IC50 values against 5Fu and gemcitabine ware measured by MTT assay. As In vivo assays, the growth of the xenografts in each cell lines was also measured for a month. The immunohistochemistry (IHC) of E-cadherin and vimentin for primary tumors and the xenografts was performed to confirm the induction of epitherial-mesenchymal transition (EMT). Result: Both OCUP-A1 and OCUP-A2 were pleomorphic cells derived from ascites of anaplastic pancreatic cancer patients. The doubling time of OCUP-A1 and OCUP-A2 was 30.9 h and 20.4 h, respectively. In migration and invasion assays, relative wound density in new 2 cell lines ranged from 47.1 % to 98.1 % at 48 hours. The values were not so different from that of other cell lines. Interestingly, although a large part of OCUP-A1 is spindle shape in normoxia, the subpopulation with polygonal structure has increased in hypoxia. Furthermore, OCUP-A1 rapidly proliferated in hypoxia. The IC50 values of gemcitabine for OCUP-A1 and OCUP-A2 were 8.77 nM and 4.90 nM, respectively. And the values of 5-Fu for OCUP-A1 and OCUP-A2 were 48.5 μM and 14.8μM, respectively. Chemosensitivity of 2 established cell lines was not superior to other cell lines. The xenografts of OCUP-A1 and OCUP-A2 more rapidly increased than other cell lines (MIAPaCa2 and Panc-1). In IHC, the E-cadherin expression was reduced and the vimentin expression was enhanced in both specimens. Conclusion: We established 2 anaplastic pancreatic cancer cell lines. It was suspected that these cell lines would have aggressive characters affected by TME and EMT. Citation Format: Kotaro Miura, Kenjiro Kimura, Ryosuke Amano, Sadaaki Yamazoe, Keiichiro Hirata, Masatsune Shibutani, Katsunobu Sakurai, Hisashi Nagahara, Takahiro Toyokawa, Naoshi Kubo, Hiroaki Tanaka, Kazuya Muguruma, Hiroshi Otani, Masakazu Yashiro, Kiyoshi Maeda, Masaichi Ohira, Kosei Hirakawa. Establishment and characterization of two cell lines of anaplastic pancreatic cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3924. doi:10.1158/1538-7445.AM2014-3924
    Type of Medium: Online Resource
    ISSN: 0008-5472 , 1538-7445
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2014
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  • 9
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 75, No. 15_Supplement ( 2015-08-01), p. 5276-5276
    Abstract: Introduction: Anaplastic pancreatic cancer (APC) is a rare subtype of pancreatic ductal adenocarcinoma (PDA). The prognosis of APC is poorer than that of ordinary PDA. There is no established therapeutic strategy as well as PDA. In this study, we compared the expression of molecular markers in intratumoral different lesions which are undifferentiated lesion (UL) and ductal lesion (DL) to reveal progression pattern of APC and hold important clue for the therapy of APC. Materials & Methods: Formalin fixed paraffin embedded blocks were made from the primary APC tissues obtained from 6 patients. Each block was manipulated for staining with Hematoxylin and Eosin (H & E) and immunohistochemistry (IHC) for SOX9, E-cadherin, vimentin, ZEB-1, Snail, N-cadherin, CD24 and CD44. Using H & E slice, each sample was divided from cancerous lesion and non-cancerous lesion. Furthermore, cancerous lesion were divided from DL and UL. And then, for each marker, the intensity of staining of UL was compared with that of DL. The intensity was scored from 0 to 3. The relation between DL and UL was estimated based on the intensity score of each sample. Results: The proportion of DL to all cancerous lesion was 0.5% to 32%. The score of SOX9 was all 3 except one case in both lesions. Although the expression of E-cadherin was almost negative in UL, DL showed various scores. About vimentin, the intensity in UL was largely 2 and 3. In contrast, that of DL was very weak or none. The score of ZEB1 was 0 or 1 in DL and 1 or 3 in UL. Although Snail showed various scores in both regions, the score was mostly higher in UL than in DL. The score of N-cadherin expression was comparatively high in UL. CD24 and CD44 were not almost expressed in DL. In UL, the expression of these markers were various intensity. Disscusion: In the current study, it was suggested that APC might be derived from PDA by showing the expression of SOX9. Previous studies were reported from the view of KRAS mutation and cytokeratin expression about the origin of APC. However, recent studies were reported that these markers were often expressed even in other pancreatic neoplasm. Furthermore the expression of SOX9 is only identified in PDA among pancreatic neoplasms. Also, it was suggested that UL might change from DL through EMT by confirming the difference of the expression pattern of markers relating EMT in DL and UL. Because the expression of CD24 and CD44 varied in each case, it is unclear whether the stemness concerned with the progression to APC in our study. We also need to estimate the other stem cell markers including CD133, Oct3/4 or Nanog. Conclusion: It was suggested that EMT might induce the progression from PDA to APC. Citation Format: Kotaro Miura, Kenjiro Kimura, Ryosuke Amano, Sadaaki Yamazoe, Go Ohira, Kohei Nishio, Masatsune Shibutani, Katsunobu Sakurai, Hisashi Nagahara, Takahiro Toyokawa, Naoshi Kubo, Hiroaki Tanaka, Kazuya Muguruma, Hiroshi Otani, Masakazu Yashiro, Kiyoshi Maeda, Masaichi Ohira, Kosei Hirakawa. Epithelial mesenchymal transition may contribute to anaplastic change of pancreatic ductal adenocarcinoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5276. doi:10.1158/1538-7445.AM2015-5276
    Type of Medium: Online Resource
    ISSN: 0008-5472 , 1538-7445
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2015
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  • 10
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 76, No. 14_Supplement ( 2016-07-15), p. 5050-5050
    Abstract: The loss of the adhesion molecules is reported to correlate with tumor invasion and metastasis in patients with various cancers. E-cadherin is an important molecule for cell-to-cell adhesion. While, CD44 is important for cell-to-extra cellular matrix adhesion. The aim of the present study is to evaluate the significance of the expression of E-cadherin and CD44 in patients with the unresectable metastatic colorectal cancer who are undergoing palliative chemotherapy. Formalin-fixed, paraffin-embedded samples were obtained from 49 patients who underwent primary tumor resection and who were receiving palliative chemotherapy for unresectable metastatic colorectal cancer. We evaluated the expression of E-cadherin and CD44 by immunohistochemistry. The expression of E-cadherin was not significantly associated with progression-free survival (PFS) or overall survival (OS). Although, the expression of CD44 did not correlate with PFS, it tended to correlate with the OS (p = 0.0699). According to the combination of CD44 and E-cadherin, both low expression group showed a significant poor PFS (p = 0.0101) and OS (p = 0.0009). Objective response rate and disease control rate were significantly decreased in both low expression group (p = 0.0076 and p = 0.0294, respectively). A univariate analysis to determine the variables that were associated with PFS indicated that the both low expression of E-cadherin and CD44 (p = 0.0474) and gender (p = 0.0330) were significantly associated with decreased survival. A multivariate analysis showed that the both low expression of E-cadherin and CD44 was an independent risk factor for decreased PFS (hazard ratio [HR]: 8.276, 95% confidence interval [CI] : 1.383-43.311; p = 0.0227). According to OS, the both low expression of E-cadherin and CD44 expression was revealed a prognostic factor by univariate and multivariate analysis(HR:15.118, 95%CI: 2.645-77.490; p = 0.0039). This study suggested that the combined low expression of both E-cadherin and CD44 was a strong independent predictor of decreased chemotherapeutic outcome and survival in patients with unresectable metastatic colorectal cancer. Citation Format: Yasuhito Iseki, Masatsune Shibutani, Kiyoshi Maeda, Hisashi Nagahara, Tatsuro Tamura, Go Ohira, Sadaaki Yamazoe, Katsunobu Sakurai, Kenjiro Kimura, Takahiro Toyokawa, Ryosuke Amano, Naoshi Kubo, Hiroaki Tanaka, Kazuya Muguruma, Masaichi Ohira, Kosei Hirakawa. The significance of the expression of E-cadherin and CD44 in patients with unresectable metastatic colorectal cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5050.
    Type of Medium: Online Resource
    ISSN: 0008-5472 , 1538-7445
    RVK:
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2016
    detail.hit.zdb_id: 2036785-5
    detail.hit.zdb_id: 1432-1
    detail.hit.zdb_id: 410466-3
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