Search

Exosomes secreted by various cells in the tumour microenvironment have been reported to be mediators of intercellular communication that play an important role in cancer progression. In this study, we aimed to investigate the effects of exosomes derived from cancer-associated fibroblasts (CAFs) on the proliferation of malignant melanoma (MM) cells and evaluated their clinicopathological significance.Three malignant melanoma cell lines, A375, MMAc, and COLO679, and three CAFs established from malignant melanomas at stages 1a, 2b, and 3b, were used. The expression of CD9, CD63, and CD81 in CAF-derived exosomes was examined using western blotting. The effect of exosomes on the proliferative potential of cancer cells was analysed using cell counting and MTT assays. The expression of CD9, CD63, and CD81 was also immunohistochemically analysed in 90 malignant melanoma specimens.CAF-derived exosomes were positive for CD9 and CD63 and remarkably inhibited the proliferative capacity of A375 and MMAc cells. The five-year disease-free survival was significantly better in patients with CAF-derived CD9-positive exosomes than in CD9-negative patients.CAF-derived exosomes, especially CD9-positive exosomes, have an inhibitory effect on the proliferation of malignant melanoma cells. These findings suggest that CD9 expression in CAFs is a promising prognostic marker for patients with malignant melanoma.

Introduction: Pelvic exenteration is a highly invasive procedure, with a reported rate of 20%–80% for serious complications. Thus, the prevention of postoperative complications is a major issue. Many previous reports have emphasized the importance of filling the dead space to prevent postoperative complications. In addition to filling the dead space, we believe that achieving sufficient drainage is extremely important. In this paper, we present a new combined method of the extended rectus abdominis myocutaneous flap and vacuum drainage with multiple drains, which achieved a lower frequency of complications. Case Presentation: The subjects were 6 patients who underwent reconstruction following pelvic exenteration during a 7-year period between April 2005 and September 2013. We retrospectively measured the volume of the pelvic cavity and that of the rectus abdominis flap on lateral computed tomography (CT) scans to calculate the percentage of the dead space that was filled by the flap. There were no problems with flap engraftment in any of the patients. There were no serious complications, and no patient required additional surgery. The percentage of the dead space filled ranged from 25% to 46% (mean: 32%). Conclusion: The rectus flap + vacuum drainage method, which uses an extended rectus abdominis myocutaneous flap to decrease the pelvic dead space and multiple vacuum suction drains, was associated with the prevention of serious complications.

Background The lymphocyte-to-monocyte ratio (LMR) has been used as a parameter reflecting systemic inflammation in several tumors, and is reportedly associated with prognosis in cancer patients. In this study, we evaluated the predictive value of LMR for progression and chemosensitivity in breast cancer patients treated with preoperative chemotherapy. Methods LMR was evaluated in 239 patients with breast cancer treated with neoadjuvant chemotherapy (NAC) with 5-fluorouracil, epirubicin, and cyclophosphamide, followed by weekly paclitaxel with or without trastuzumab, and subsequent curative surgery. The correlations between LMR and clinicopathological features, prognosis, and pathological complete response (pCR) rate of NAC were evaluated retrospectively. We also evaluated the predictive value of neutrophil-to-lymphocyte ratio (NLR), and compared the predictive values of LMR and NLR. Results We set 6.00 as the cut-off level for LMR based on the receiver operating characteristic (ROC) curve. A total of 119 patients (49.8%) were classified in the high-LMR group and 120 (50.2%) were classified in the low-LMR group. The low-LMR group had significantly worse disease-free survival rate (DFS) in all patients (p = 0.005) and in triple-negative breast cancer patients (p = 0.006). However, there was no significant correlation between LMR and pCR. Multivariate analysis showed that low LMR was an independent risk factor for DFS (p = 0.008, hazard ratio = 2.245). However, there was no significant difference in DFS (p = 0.143, log-rank) between patients in the low- and high-NLR groups. Conclusions LMR may be a useful prognostic marker in patients with breast cancer. Electronic supplementary material The online version of this article (10.1186/s12885-018-5051-9) contains supplementary material, which is available to authorized users.

The surgical resection of a primary melanoma is sometimes followed by the immediate development of distant metastases, suggesting that the primary melanoma might control the metastatic process. We hypothesized that a paracrine factor(s) from primary melanoma cells might regulate the progression of metastasizing melanoma cells. Here we attempted to identify the factor(s) from primary melanoma cells that regulate the invasion ability of metastatic melanoma cells. We used two mouse melanoma cell lines, B16 and B16/BL6, that latter of which is a subline of B16 melanoma and shows high metastatic potential to lung. We investigated the interaction between the parent B16 cells and daughter B16/BL6 cells by invasion assay, cell morphology, cytokine array, RT-PCR, and gelatin-zymography. The conditioned medium (CM) from B16 significantly (p=0.02) inhibited the invasion ability of B16/BL6 cells. The morphology of the B16/BL6 cells was changed from bipolar shape to a multipolar shape following the addition of the CM from B16. The B16 cells produced high levels of C-X-C motif ligand 1 (CXCL1), CXCL10, and M-CSF compared to the B16/BL6 cells. CXCL1 significantly (p=0.01) decreased the invasion ability of B16/BL6 cells, but CXCL10 and M-CSF did not. The invasion-inhibitory activity of the CM from B16 was significantly (p=0.046) suppressed following the addition of a neutralizing anti-CXCL1 antibody. The CM of B16 and CXCL1 increased the E-cadherin mRNA level and decreased MMP2 activity of B16/BL6 cells. These findings suggested that primary melanoma cells might down-regulate the invasion activity of metastatic melanoma cells through CXCL1 signaling.

Background Immune responses in a tumour microenvironment can be evaluated by analysing tumour-infiltrating lymphocyte (TIL) density; this has been verified in the clinical setting. Although there are many reports on TIL density in primary tumours, little is known about its density in recurrent tumours. Methods Of 300 patients treated with neoadjuvant chemotherapy during the study period, 29 were considered for evaluation of TIL density in primary and recurrent tumours. We performed a retrospective analysis of the association between TIL density and prognosis. Results TIL density was significantly lower in recurrent tumours than in primary tumours (P = 0.007). There was no correlation between post-recurrence survival and TIL density in core-needle biopsy specimens obtained from primary tumours (P = 0.837). However, patients with high TIL density in recurrent tumours had significantly better post-recurrence survival than did the corresponding group with low TIL density (P = 0.041). Multivariate analysis revealed that high TIL density contributed significantly towards improving post-recurrence survival in all patients (P = 0.035; hazard ratio, 0.167). Conclusions In recurrent breast cancer, a decrease in TILs density was observed as compared to the primary tumour, and this affects the poor prognosis after relapse.

Background:. The quality of life of the face involves mainly its configuration, and it plays an important functional role in communication skills. Thus, having artificial eyes is extremely essential in maintaining the quality of life of patients who have lost their eyeballs. We will present the details of the technique, including tips and innovations for eye socket reconstruction using spherical costal cartilage implant, which leads to dynamic and aesthetic results. Methods:. There were 19 cases of eye socket reconstruction using costal cartilages from 2008 to 2020. The patient age range was 18–77 years old. There were 10 cases of anophthalmia and 9 cases of ocular phthisis. In our operative method, we created the costal cartilage implant by harvesting the sixth costal cartilage of the affected side. If extraocular muscle remained, we sutured each muscle to the cartilage. Results:. Our method made application of thin artificial eyes possible in all cases. Regarding postoperative complications, there were 1 case of postoperative infection and 1 case of vascular failure of temporoparietal fascial flap. Seventeen cases were wet socket, and 2 cases were dry socket. We had attained movement of the artificial eye in 15 cases. Conclusions:. Eye socket reconstruction is considered one of the most challenging operations, and various postoperative complications appear in the long term. Costal cartilages are considered as the most suitable materials to create the base of artificial eyes.

Introduction: Cancer-associated fibroblasts (CAFs) in tumor microenvironment have an interaction with tumor cells and play an important role for the progression and metastasis. Tumor lymphangiogenesis may be controlled by not only tumor cells but also tumor microenvironment, and some reports suggested that CAFs may promote tumor lymphangiogenesis. In malignant melanoma, lymph node metastasis is important prognostic factor. However, no study about the interaction among CAFs, tumor cells, and human lymphatic endothelial cells (HLECs) was found. In this study, we evaluated the effect of CAFs on lymphatic vessel formation in malignant melanoma. Materials and methods: We used CAFs established from a primary malignant melanoma, HLECs, and two melanoma cell lines (MMAc, COLO679). CAFs and melanoma cell lines were cultured with DMEM. Serum-free conditioned medium (CM) was obtained from CAFs (CAF-CM) and two melanoma cells (MMAc-CM, COLO679-CM). Also, CM from CAFs cultured with MMAc-CM (MMAcCAF-CM) and COLO679-CM (COLO679CAF-CM) was obtained. HLECs were cultured by endothelial growth medium (EGM) in combination with 50% of each CM. Control group was cultured with 50% EGM and 50% of serum-free endothelial basal medium. The effect of each CM on the proliferation of HLECs was examined by CCK assay and tube formation assay. Tube formation was assessed by counting total tube length, number of tubes and junctions at 12 hours of incubation. Result: Both MMAcCAF-CM and COLO679CAF-CM significantly increased the proliferation of HLECs in compared with the control or CAF-CM alone, after 24, 48 and 72 hours of incubation. In tube formation assay, both tube length and number of tubes formed by HLECs were significantly longer and larger in MMAcCAF-CM and MMAc-CM group than control group. The tube length, number of tubes and junctions treated by MMAcCAF-CM or COLO679CAF-CM were longer and larger than in control and CAF-CM group. Conclusion: CAFs under the influence of melanoma cells might have a more progressive effect on HLECs, resulting in promotion of lymphangiogenesis. Citation Format: Shusaku Maeda, Masakazu Yashiro, Hisashi Motomura, Takaharu Hatano, Heishiro Fujikawa. Cancer-associated fibroblasts under the influence of melanoma cells might stimulate the lymphatic vessel formation [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5115.

Background: Studies have found that patients with cancer exhibit abnormal leukocyte fractions, such as elevated neutrophil count and diminished lymphocyte count, and that the neutrophil-to-lymphocyte ratio (NLR) provides a surrogate marker for prognosis and response to treatment of patients after radical surgery for several different types of cancer. However, few reports have addressed the association between the NLR and response to endocrine therapy. In this study, we carried out a clinical investigation to confirm whether or not the NLR predicted the response to endocrine therapy of stage IV breast cancer. Patients and Methods: The study subjects were 34 patients who underwent endocrine therapy as initial drug therapy for stage IV breast cancer. The correlation between NLR and prognosis, including the efficacy of endocrine therapy, was evaluated retrospectively. Results: Among the 34 patients, the NLR was high in 10 (29.4%) and low in 24 (70.6%). In analysis of outcomes, the group with low NLR had a significant prolongation of progression-free survival (p=0.003), time to treatment failure (p=0.031), and overall survival (p=0.013) compared to the group with high NLR. Univariate analysis of progression-free survival found that responding to treatment [hazard ratio (HR)=4.310, p=0.004] and low NLR (HR=3.940, p=0.016) were factors associated with a favorable prognosis. Multivariate analysis also showed that responding to treatment (HR=4.329, p=0.006) and low NLR (HR=3.930, p=0.008) were independent factors associated with a favorable prognosis. Conclusion: Our results suggested that the NLR may represent a predictive marker for response to endocrine therapy in stage IV breast cancer.

Background Circulating tumor cells (CTCs) are linked to metastatic relapse and are regarded as a prognostic marker for human cancer. High expression of plakoglobin, a cell adhesion protein, within the primary tumor is positively associated with CTC clusters in breast cancer. In this study, we investigated the correlation between plakoglobin expression and survival of breast cancer. Methods We evaluated 121 breast cancer patients treated with neoadjuvant chemotherapy. Expression of plakoglobin was identified by immunohistochemical staining in the cell membrane. We also examined the relation between the expression of plakoglobin and E-cadherin, an epithelial–mesenchymal transition (EMT) marker. Results Patients with high plakoglobin expression had significantly worse distant-metastasis-free survival (DMFS) (P = 0.016, log rank). Plakoglobin expression had no correlation with pathological complete response rate (P = 0.627). On univariate analysis with respect to distant metastasis, high plakoglobin expression showed worse prognosis than low plakoglobin expression [P = 0.036, hazard ratio (HR) = 3.719]. Multivariate analysis found the same result (P = 0.013, HR = 5.052). In addition, there was a significant relationship between the expression of plakoglobin and E-cadherin (P = 0.023). Conclusions Plakoglobin expression is an independent prognostic factor in patients with breast cancer, particularly for DMFS, and this is related to EMT. Electronic supplementary material The online version of this article (doi:10.1186/s40364-017-0099-2) contains supplementary material, which is available to authorized users.

The rate of local recurrence after mastectomy is reportedly similar to that of one-stage reconstruction. Most recurrences are in the skin or chest wall, while recurrence in the reconstructed breast is rare and the causes are uncertain. We report a case of a 42-year-old female who underwent partial mastectomy for left breast cancer with cT4aN0M1(PUL), cStage IV after endocrine therapy 3 years ago. Histopathological diagnosis was solid-tubular carcinoma. She had been treated with only endocrine therapy but diagnosed with local recurrence in the left breast. She underwent total mastectomy and rectus abdominis musculocutaneous flap reconstruction. Partial flap necrosis occurred following conservative therapy. She was accordingly treated with anastrozole and GnRH agonist. A mass approximately 1.5 cm in size was palpated inside of the reconstructed breast. As such, she was diagnosed with recurrence in the reconstructed breast through ultrasound biopsy. She underwent partial resection of the left precordial tumor, and histopathological examination revealed scirrhous cancer. She is currently well without any recurrence.

Background Tumour-infiltrating lymphocytes (TILs) can be used to monitor the immune tumour microenvironment (iTME) and predict treatment response and outcome in breast cancer. We evaluated the prognostic significance of the levels of CD8+ TILs and forkhead box protein (FOXP3)-positive TILs before and after neoadjuvant chemotherapy (NAC). Patients and methods We examined 136 patients with breast cancer treated with NAC. The number of CD8+ TILs and FOXP3+ TILs in biopsy specimens and residual tumours was evaluated by immunohistochemistry. Results Patients with a high rate of change in the CD8/FOXP3 ratio (CFR) had significantly better recurrence-free survival (RFS) (p Conclusion Improvement in the iTME following NAC is correlated with good outcome. The rate of change in the CFR may be a useful biomarker to predict prognosis of patients treated with NAC.

BACKGROUND/AIM Recent interest has focused on the significance of tumor-infiltrating lymphocytes (TILs) on the efficacies and outcomes of the treatment in breast cancer (BC). Based on the recent international recommendation to standardize the evaluation method, the clinical validity and utility of TILs in patients who underwent neoadjuvant chemotherapy (NAC) were investigated in the present study. PATIENTS AND METHODS TILs were evaluated in 177 patients with BC treated with NAC and subsequent curative surgery. The correlation between TILs evaluated according to the standard method and prognosis, including the efficacy of NAC, was investigated retrospectively. RESULTS In the high-TIL group (n=96) compared to the low-TIL group (n=81), triple-negative breast cancer (TNBC) (p

Background The tumor immune environment not only modulates the effects of immunotherapy, but also the effects of other anticancer drugs and treatment outcomes. These immune responses can be evaluated with tumor-infiltrating lymphocytes (TILs), which has frequently been verified clinically. On the other hand, residual cancer burden (RCB) evaluation has been shown to be a useful predictor of survival after neoadjuvant chemotherapy (NAC). In this study, RCB and TILs evaluations were combined to produce an indicator that we have termed “RCB-TILs”, and its clinical application to NAC for breast cancer was verified by subtype-stratified analysis. Methods A total of 177 patients with breast cancer were treated with NAC. The correlation between RCB and TILs evaluated according to the standard method, and prognosis, including the efficacy of NAC, was investigated retrospectively. The RCB and TILs evaluations were combined to create the “RCB-TILs”. Patients who were RCB-positive and had high TILs were considered RCB-TILs-positive, and all other combinations were RCB-TILs-negative. Results On multivariable analysis, being RCB-TILs-positive was an independent factor for recurrence after NAC in all patients (p < 0.001, hazard ratio = 0.048), triple-negative breast cancer (TNBC) patients (p = 0.018, hazard ratio = 0.041), HER2-positive breast cancer (HER2BC) patients (p = 0.036, hazard ratio = 0.134), and hormone receptor-positive breast cancer (HRBC) patients (p = 0.002, hazard ratio = 0.081). Conclusions The results of the present study suggest that RCB-TILs is a significant predictor for breast cancer recurrence after NAC and may be a more sensitive indicator than TILs alone. Electronic supplementary material The online version of this article (doi: 10.1186/s12885-017-3927-8) contains supplementary material, which is available to authorized users.

Background/aim Eribulin mesylate (eribulin) is currently used for the treatment of locally advanced or metastatic breast cancer (MBC). It is a cytotoxic agent with unique mechanisms that suppress the epithelial-mesenchymal transition (EMT) of cancer cells and promote tumor vascular remodeling. In this study, we investigated the expression of markers for EMT and hypoxia in sets of clinical specimens collected before and after eribulin treatment to verify its unique mechanisms. Patients and methods The expression of markers for EMT and cellular hypoxia [E-cadherin, N-cadherin, vimentin, and carbonic anhydrase 9 (CA9)] was examined immunohistochemically in MBC tissues collected from 20 patients before and after chemotherapy with either eribulin (n=10) or paclitaxel (n=10). Results An increase of E-cadherin and decrease of CA9 expression were observed in MBC tissues from patients with objective clinical responses to eribulin treatment. Patients with E-cadherin-positive conversion and CA9-negative conversion had significantly higher response rates (p=0.004 and p=0.024, respectively) and prolonged time to treatment failure (p=0.018 and p=0.038, respectively) than patients without changes in marker expression. Conclusion Expression of EMT and hypoxia markers in clinical samples from patients with MBC was suppressed by eribulin treatment. The results provide additional clinical data on improved survival of patients treated with eribulin and the mechanism of response.

Background/aim Recently, reports of the clinical implications of tumor-infiltrating lymphocytes (TILs) in breast cancer treatment have increased. We evaluated that chemotherapy with a TPD regimen (trastuzumab, pertuzumab, docetaxel) against HER2-positive breast cancer, using TILs as indicators. Patients and methods The subjects were 24 patients who had received TPD-chemotherapy. A semi-quantitative evaluation of lymphocytes invading the stroma in needle biopsy specimens prior to treatment as TILs was conducted, after which, sensitivity to chemotherapy and patient prognosis were evaluated. Results Overall response rate was significantly higher in the high-TILs group than in the low-TILs group. Significant extension of the progression-free survival (PFS) and overall survival was found in the high-TILs group compared to the low-TILs group. In addition, high TILs numbers significantly contributed to the extension of PFS. Conclusion Monitoring antitumor immune response using TILs might be a useful indicator for predicting the curative effects of TPD chemotherapy for HER2-positive breast cancer.

We present a modified locoregional flap for the reconstruction of large anterior skull base defects that should be reconstructed with a free flap according to Yano's algorithm. No classification of skull base defects had been proposed for a long time. Yano et al suggested a new classification in 2012. The lb defect of Yano's classification extends horizontally from the cribriform plate to the orbital roof. According to Yano's algorithm for subsequent skull base reconstructive procedures, a lb defect should be reconstructed with a free flap such as an anterolateral thigh free flap or rectus abdominis myocutaneous free flap. However, our modified locoregional flap has also enabled reconstruction of lb defects. In this case series, we used a locoregional flap for lb defects. No major postoperative complications occurred. We present our modified locoregional flap that enables reconstruction of lb defects.

Introduction: Malignant tumors are composed of cancer cells and various types of stromal cells, including fibroblasts, vascular endothelial cells, and immune cells. It has been reported that fibroblasts in the tumor stroma termed as cancer-associated fibroblasts (CAFs) may affect cancer progression. However, the currently available evidence regarding the function of CAFs remains controversial, with some studies reporting stimulatory activity and some studies reporting inhibitory activity in malignant melanoma cells. Previously, we reported that CAFs possibly exert a stimulatory effect on the proliferation of melanoma cells. In this study, we identified a factor from CAFs that are involved in melanoma progression. Materials and Methods: We used three malignant melanoma cell lines, namely MMAc, COLO679, and A375. Three CAFs and one type of non-affected fibroblasts (NAFs) were established from sebaceous carcinoma/skin malignant melanoma and normal skin, respectively. Serum-free conditioned medium from CAFs (CAF-CM) was prepared as follows: CAFs were seeded into 100-mm plastic dishes with 10 ml of Dulbecco's modification of Eagle medium (DMEM) containing 5% fetal calf serum. To obtain CAF-CM, CAFs were washed using phosphate-buffered saline and subsequently incubated for 3 days with serum-free DMEM, and the conditioned medium was collected as CAF-CM. Additionally, we obtained NAF-CM from NAFs using the same procedure. These fibroblasts were cultured and used before the 15th passage. The cytokines included in each conditioned medium were determined using cytokine assay. Subsequently, we selected some cytokines, which were released more abundantly in CAFs than in NAFs, as potential factors affecting the proliferation of melanoma cells. The effects of these cytokines on the proliferation of melanoma cells were determined using MTT assay and cytometry assay using a Coulter Counter. Results: CAF-CM contained significantly high concentration of MIP-3β, in compared with that in NAF-CM. In addition, MIP-3β significantly increases in the proliferation of melanoma cells, A375 and COLO679, at 48 h and 72 h culture. Conclusion: CAFs from skin cancer might promote the proliferation of malignant melanoma cells via MIP-3β signaling. Citation Format: Heishiro Fujikawa, Masakazu Yashiro, Takaharu Hatano, Shusaku Maeda, Hisashi Motomura. Macrophage Inflammatory Protein-3 beta (MIP-3β) from cancer-associated fibroblasts promote the proliferation of melanoma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2037.

Introduction: It has been reported that cancer-associated fibroblasts (CAFs) in tumor microenvironment may play an important role for the progression and metastasis of cancer cells. Some reports suggested that CAFs may promote tumor lymphangiogenesis. In malignant melanoma, lymph node metastasis is one of important prognostic factors. Tumor lymphangiogenesis may be controlled not only by tumor cells but also tumor microenvironment such as CAFs, however no study about the interaction between CAFs and human lymphatic endothelial cells (HLECs) was found. Then, we evaluated the effect of CAFs on lymphatic vessel formation in malignant melanoma. Materials and methods: We used CAFs, normal tissue associated fibroblasts (NAFs) established from a patient of malignant melanoma, and HLECs. Serum-free conditioned medium was obtained from NAFs (NAF-CM) or CAFs (CAFs-CM). CAFs and NAFs were cultured with or without supernatant from malignant melanoma cells. HLECs were cultured by EBM with 50% of NAF-CM or CAFs-CM. Control group was cultured with 50% EBM and 50% of serum-free medium. The effect of each CM on the proliferation of HLECs was examined by CCK assay and tube formation assay. Tube formation was assessed at 12 hours of incubation by fluorescence microscope. Result: CAFs-CM significantly decreased the proliferation of HLECs in comparison with NAFs-CM after 48, 72 and 96 hours of incubation by CCK assay. In tube formation assay, tube length and number of tubes and junctions were also significantly shorter and smaller in CAFs group, in compared with the control and NAFs group. Conclusion: CAFs might have a suppressive effect on HLECs, resulting in inhibition of lymphangiogenesis. Citation Format: Shusaku Maeda, Masakazu Yashiro, Hisashi Motomura, Takaharu Hatano, Heishiro Fujikawa. Effect of cancer-associated fibroblasts on lymphatic vessel formation in malignant melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3763.

Summary:. The treatment for arterial venous malformation (AVM) of functionally and aesthetically important parts such as the hand is considered to be challenging. AVM existed in the right forearm and the thenar region of 55-year-old man. Combined method with free omentum flap and split-thickness skin flap was performed for hand reconstruction after radical excision of an AVM. The postoperative course was good. Seven years have passed postoperatively, and functional but aesthetical results are satisfying with no recurrence. Our operative procedure with complete resection of AVM with placement of the free omental flap to the resected area and placing back the original skin as a skin graft is considered to be an ideal curative surgical treatment of the AVM.

Object In 1997 the authors presented the case of a 3-year-old girl who underwent complete resection of a huge tumor via a new technique using a dismasking flap. Since that report, 14 patients have undergone surgery using this technique. There have been few reports on long-term follow-up after a craniofacial surgical approach and reconstruction of a huge tissue defect. The authors report details of this procedure based on these cases, including long-term follow-up in the original case. Methods The dismasking flap is a cranioorbitofacial degloving method that uses a circumpalpebral approach with or without piriform margin incisions and presents a wide surgical field under direct vision. Fourteen patients have undergone surgery using the dismasking flap technique. These patients had multiple craniofacial bone fractures (3 cases) and malignant or premalignant craniofacial tumors (11 cases). Patients ranged in age from 3 to 62 years old. The longest follow-up period was 15 years. The results of these cases are reported regarding changes in the facial bones and soft tissues with growth, the various pathologies involved, and complications. Results Complications using this approach were lagophthalmos with ectropion of the lower eyelid, disturbance of the superior orbital nerve, disturbance of the inferior orbital nerve, maxillary hypoplasia, and blepharoptosis. In all affected patients these complications were almost always temporary. In 2 of the 14 cases, a repair operation for lagophthalmos was necessary, while others showed spontaneous improvement approximately 6 months to a year after the procedure. In the case with the longest follow-up duration, the patient's nose is asymmetrical, especially the alar portion, due to hypoplasia of the grafted bone and/or the fixation with titanium miniplates. Conclusions This flap is very useful for en bloc resection of huge skull base tumors, multiple craniofacial bone fractures, and as a lateral approach to a deep portion of the middle cranial base. Careful attention is necessary, however, because one may encounter unexpected complications if one does not ensure adequate protection of the perioperative flap. Meticulous postoperative management is also essential.

The functional and cosmetic results of the reconstructive surgery after extended total maxillectomy greatly depend on the quality of the orbital reconstruction. In order to achieve good orbital reconstruction, we developed the dynamic eye socket reconstruction using temporalis transfer. In this report, I will present the details of the technique, including tips and innovations for dynamic eye socket reconstruction.Two patients (2 males, aged 70 and 72 years old) who underwent extensive resection of midfacial tumours were treated with dynamic eye socket reconstruction using temporalis transfer. The follow up period was 16 and 102 months. No acute complications were observed. The movements of the upper/lower eyelids including crow's feet were observed and a good shape in the reconstructed medial/lateral canthal area was maintained in all patients.This procedure provides both the eyelids with movement and also a good shape in the reconstructed medial/lateral canthal region. Furthermore, it contributes to achieving satisfactory functional and cosmetic results in the orbital reconstruction.

Introduction Malignant solid tumor is composed of not only cancer cells but also various types of stromal cells, which include fibroblasts, vascular endothelial cells, and immune cells. Currently, it has been reported that fibroblasts in tumor stroma, namely cancer-associated fibroblasts (CAFs), might affect cancer progression. Some studies suggested that CAFs have the stimulation-activity of the malignant potential of melanoma cells, but some reports mentioned their inhibitory-activity of melanoma cells. In this study, we investigated the effects of CAFs on the proliferation and invasion ability of malignant melanoma cells. Material and Method We used 2 malignant melanoma cell lines, MMAc and COLO679. Three CAFs were established from sebaceous carcinoma or skin malignant melanoma. Serum-free conditioned medium (CAF-CM) from CAF was prepared as follows. CAF were seeded into 100 mm plastic dishes with 10 ml of DMEM containing 2% FCS. To obtain CAF-CM, CAF were washed by PBS and then incubated for 3 days with serum-free DMEM. The conditioned medium was collected as CAF-CM. Serum-free DMEM was used as the control. CAFs were used before the 15th passage in culture. The effect of CAF-CM on the proliferation of the melanoma cells was determined by cytometry assay using Coulter Counter. The effect of CAF-CM on the motility of the melanoma cells was examined by Matrigel-invasion assay. Result CAF-CM significantly increased the proliferation of melanoma cell, in compared the control. Under the condition of adding 25% of CAF-CM, the number of MMAc cells were increased about 300% after 72 hours of incubation, and 105% after 120 hours of incubation that of COLO679 increased about 56% after 72 hours of incubation, 130% after 120 hours of incubation. CAF-CM significantly increased the invasion of MMAc cells. The invasion of MMAc cells were increased about 200% after 72 hours of incubation following the addiction of 25% CAF-CM. In contrast, CAF-CM did not affect on the invasion of COLO679 cells. Conclusion CAFs are associated with the proliferation and invasion of malignant melanoma cells. Citation Format: Heishiro Fujikawa, Masakazu Yashiro, Takaharu Hatano, Shusaku Maeda, Hisashi Motomura. Effect of cancer-associated fibroblasts on the progression of malignant melanoma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4489.

[Introduction]The prognosis of patients with malignant melanoma is extremely poor because of the frequent metastasis. We have occasionally experienced the clinical cases that malignant melanoma patients rapidly developed distant metastases after the surgical resection of primary cancer lesion. These cases suggested that a primary tumor may control a metastatic tumor. We hypothesized that primary melanoma cells might produce a factor(s) which regulate the progression of metastatic melanoma cells. We reported that the conditioned medium from B16 cells significantly (p=0.02) suppressed the invasion ability of B16/BL6 cells, which is a subline of B16 melanoma and shows high metastatic potential to lung at the previous meeting. Then we investigated the growth and invasion interaction between primary malignant melanoma cells and high metastatic melanoma cells at the molecular level. [Materials and methods]We used two mouse melanoma cell lines, B16 and B16/BL6, that latter of which is a subline of B16 melanoma and shows high metastatic potential to lung. We investigated the interaction between the parent B16 cells and daughter B16/BL6 cells by invasion assay, cell morphology, cytokine array, RT-PCR, and gelatin-zymography. [Results]The conditioned medium (CM) from B16 cells significantly (p=0.02) inhibited the invasion ability of B16/BL6 cells. The morphology of the B16/BL6 cells was changed by the CM from bipolar in B16 cells to a multipolar shape. The B16 cells produced high levels of C-X-C motif ligand 1 (CXCL1), CXCL10, and M-CSF compared to the B16/BL6 cells. CXCL1 significantly (p=0.01) decreased the invasion ability of B16/BL6 cells, but CXCL10 and M-CSF did not. The invasion-inhibitory activity of the CM from B16 cells was significantly (p=0.046) suppressed following the addition of a neutralizing anti-CXCL1 antibody. The CM of B16 and CXCL1 increased the E-cadherin mRNA level and MMP2 activity of B16/BL6 cells. [Conclusion]Primary melanoma cells might down-regulate the invasion activity of metastatic melanoma cells through CXCL1 signaling. Citation Format: Takaharu Hatano, Masakazu Yashiro, Hisashi Motomura, Heishiro Fujikawa, Shusaku Maeda. C-X-C motif ligand 1 from melanoma cells downregulates the invasion of their metastatic melanoma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4507.

Background & Aims: Activated fibroblasts in the tumor stroma, also termed carcinoma-associated fibroblasts (CAFs), play a critical role in the progression of cancer. In our previous studies, we identified CAFs as playing an important role in cancer progression in the development of gastric cancer. However, it remains unclear which factor recruit and promote CAFs. The aim of this study is to clarify from where CAFs originate and which factor recruits them into the gastric cancer microenvironment. Methods: In vitro study analyzed the chemotaxis-stimulating factor from gastric cancer cells using bone marrow-derived mesenchymal cells (BM-MCs). The nude mice with bone marrow transplantation from CAG-EGFP mice were used for the in vivo gastric cancer experiments. The influences of chemokine (C-X-C motif) receptor 2 (CXCR2) inhibitor on the migration of BM-MCs were examined in vitro and in vivo. Results: BM-MCs migrated into tumor stroma of in vivo gastric cancer. The number of BM-MCs was much greater in the gastric tumor than in normal tissue from three days after inoculation of cancer cells. C-X-C motif ligand 1 (CXCL1) from gastric cancer cells significantly (p Conclusions: BM-MSCs recruit into gastric tumor and differentiate into CAFs. CXCL1 from gastric cancer cells stimulates the recruitment of BM-MCs into gastric cancer locus via CXCR2 signalling. Inhibition of BM-MCs’ recruitment appears a promising therapy for gastric cancer. Citation Format: Masakazu Yashiro, Tomohisa Okuno, Hiroaki Kasashima, Yuichiro Miki, Hirohisa Nakamae, Kisyu Kitayama, Toshiyuki Kawashima, Takaharu Hatano, Heishiro Fujikawa, Tsuyoshi Hasegawa, Takahiko Nakane, Masayuki Hino, Kosei Hirakawa, Masaichi Ohira. CXCL1-CXCR2 signaling might recruit the bone marrow-derived mesenchymal cells to the carcinoma-associated fibroblasts of gastric cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 941. doi:10.1158/1538-7445.AM2017-941

Introduction: The prognosis of patients with malignant melanoma is extremely poor because of the frequent metastasis. We have occasionally experienced the clinical cases that malignant melanoma patients rapidly developed distant metastases after the surgical resection of primary cancer lesion. These cases suggested that a primary tumor may control a metastatic tumor. We hypothesized that primary melanoma cells might produce a factor(s) which regulate the progression of metastatic melanoma cells. Then we investigated the growth and invasion interaction between primary malignant melanoma cells and high metastatic melanoma cells. Materials and methods: We used two malignant melanoma cell lines, B16 as a parent primary melanoma cell line and a selected daughter cell line B16/BL6 with high metastatic ability. We investigated the invasive interaction between the parent B16 cells and daughter B16/BL6 cells in vitro. The conditioned medium from B16 cells and B16/BL6 cells was added to B16 cells or B16/BL6 cells. Then we examined the effect of these conditioned medium on the motility and proliferation of melanoma cells by MTT assay or invasion assay at 24 hours and 72 hours after the addiction of the conditioned medium. Results:The conditioned medium from B16 cells significantly (p=0.02) suppressed the invasion ability of B16/BL6 cells at 24 hours after the addiction of the conditioned medium. In contrast, the invasion ability of B16 cells were not affected by the conditioned medium from B16/BL6 cells at 24 hours. Each conditioned medium did not affect on the proliferation of both melanoma cell lines. On the other hand, at 72 hours after the addiction of the conditioned medium from B16 cells significantly (p=0.000003) suppressed the growth of B16/BL6 cells. Conclusion: Primary melanoma cells might down-regulate the progression of metastatic melanoma cells by a soluble factor(s). Citation Format: Takaharu Hatano, HIsashi Motomura, Heishiro Fujikawa, Masakazu Yashiro, Kishu Kitayama. Interaction between the malignant melanoma cell of the primary lesion and the metastatic lesion [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2985. doi:10.1158/1538-7445.AM2017-2985

The functional and esthetic results of reconstructive surgery after extended total maxillectomy or extended orbital exenteration greatly depend on the quality of the orbital reconstruction. We developed dynamic eye socket reconstruction using temporalis transfer to achieve good orbital reconstruction, and examined the usefulness of our technique. Five patients (three men and two women, aged 44-72 years) who underwent extensive resection of midfacial malignancies were treated with dynamic eye socket reconstruction using temporalis transfer. In most cases, eye socket reconstruction was performed approximately 1 year after the initial surgery, and temporalis transfer was used after maturation of the eye socket. The follow-up period ranged from 16 to 120 months (average 63.8 months). Movement of the upper and lower eyelids was achieved in all cases, and definite creases at the lateral canthus were observed in two patients. A good shape in the reconstructed medial and lateral canthal areas was maintained in all patients. Our reconstruction technique is extremely effective in creating natural creases ('crow's feet') at the lateral canthus during smiling, enabling movement of the upper and lower eyelids, and maintaining a sharp palpebral morphology.

Buerger's disease presents a difficult challenge, with its uncertain etiology and lack of a standardized therapy, and is often refractory to treatment. In the present report, we have described the case of a 43-year-old male with Buerger's disease, a refractory ulcer on his right great toe, and severe pain at rest. We obtained favorable results using a hybrid therapy that combined distal bypass with a free temporoparietal fascial flap. Postoperatively, the flap healed, and the patient's pain resolved. After more than 3 years of follow-up, he had had no recurrence of the foot ulceration, and the patency of the bypass graft had been confirmed using Doppler flowmetry. We believe that the hybrid therapy we devised for the present patient (ie, combined distal bypass with a free temporoparietal fascial flap) is an effective treatment of Buerger's disease with pedal ulceration. The therapeutic strategies for this condition should not adhere to a single treatment modality; rather, they should combine available treatment modalities according to each individual patient's condition.

In 1997 the authors presented the case of a 3-year-old girl who underwent complete resection of a huge tumor via a new technique using a dismasking flap. Since that report, 14 patients have undergone surgery using this technique. There have been few reports on long-term follow-up after a craniofacial surgical approach and reconstruction of a huge tissue defect. The authors report details of this procedure based on these cases, including long-term follow-up in the original case.The dismasking flap is a cranioorbitofacial degloving method that uses a circumpalpebral approach with or without piriform margin incisions and presents a wide surgical field under direct vision. Fourteen patients have undergone surgery using the dismasking flap technique. These patients had multiple craniofacial bone fractures (3 cases) and malignant or premalignant craniofacial tumors (11 cases). Patients ranged in age from 3 to 62 years old. The longest follow-up period was 15 years. The results of these cases are reported regarding changes in the facial bones and soft tissues with growth, the various pathologies involved, and complications.Complications using this approach were lagophthalmos with ectropion of the lower eyelid, disturbance of the superior orbital nerve, disturbance of the inferior orbital nerve, maxillary hypoplasia, and blepharoptosis. In all affected patients these complications were almost always temporary. In 2 of the 14 cases, a repair operation for lagophthalmos was necessary, while others showed spontaneous improvement approximately 6 months to a year after the procedure. In the case with the longest follow-up duration, the patient's nose is asymmetrical, especially the alar portion, due to hypoplasia of the grafted bone and/or the fixation with titanium miniplates.This flap is very useful for en bloc resection of huge skull base tumors, multiple craniofacial bone fractures, and as a lateral approach to a deep portion of the middle cranial base. Careful attention is necessary, however, because one may encounter unexpected complications if one does not ensure adequate protection of the perioperative flap. Meticulous postoperative management is also essential.

Emphasis of secondary repair for unilateral cleft-lip has been placed on correction of the cleft-lip nasal deformity by translocation of the alar cartilage with its attached vestibular lining into a normal position, thereby establishing the normal vault and shape of the cartilage. We have managed cleft-lip deformities employing a modified reverse-U method. We present our modified technique and the obtained results. Eighty-nine patients with unilateral cleft-lip nasal deformity underwent surgical repair between 1998 and 2007 by one surgeon. These patients were divided into two groups based on their previous operative histories. Group A comprised 52 patients who underwent primary cheiloplasty using a modified Tajima technique. No other surgery had been performed prior to our modified reverse-U method in these cases. Group B consisted of 37 patients receiving the primary operation and/or more than one rhinoplasty at another hospital before our method was employed. The ages of our patients at the time of the secondary operation ranged from 4 to 40 years (average, 8.1 years; 4.2 years in group A and 13.4 years in group B). The follow-up period ranged from 2 to 11 years (average 7.2 years). For objective evaluation, points on the nasal dome and alar crease were measured on patient photographs. The results were retrospectively distinguished to three levels by total point score: Excellent, Good and Fair. Excellent or Good results were obtained in 74 cases, while 15 had Fair results. Eighty-one percent of all cases maintained acceptable results without relapse. Fair results were attributed to an unclear alar crease or relapse. In conclusion, rigid fixation and release of nasal cartilage are very important. We believe our modified reverse-U method to be very useful for achieving symmetry of the unilateral cleft-lip nose in the long term.