Cancer stem cells (CSCs) play an important role in cancer development, its main functions are self-renewing capacity, chemoresistance and tumorigenic capacity. The aim of this study is to clarify the possible role of Shh signaling in regulation of CSCs. METHODS: Normal cancer cells (HCT-116) were cultured with serum medium and cancer stem-like cells (CSCs) were obtained from serum-free medium after incubation for 14 days. After cell culturing was done RNA extraction and cDNA transcription of NCs and CSCs (HCT-116). The expressions mRNA of surface markers (CD44, EpCAM), stemness genes (Oct-4, Nanog), Shh signaling (Ptch1, SMO), and shh pathway downstream gene (Gli1), EMT markers (E-Cadherin, Vimentin) and TJ genes (Claudin-4, Occludin) were determined by real time RT-PCR before and after administration of cyclopamine (2, 5 μM). RESULTS: The expressions of surface markers (CD44, EpCAM) and stemness genes (Oct-4, Nanog) were significantly highly expressed in CSCs. Shh signaling pathway Ptch1, SMO and downstream gene Gli1 were significantly higher in CSCs than in NCs. Epithelial marker E-Cadherin was reduced in CSCs, mesenchymal marker Vimentin was up-regulated in CSCs. The expressions of Claudin-4 and Occludin were significantly higher in CSCs compared with NCs. SMO, Gli1 and Vimnetin were significantly inhibited after administration of cyclopamine (2, 5μM), but E-Cadherin was up-regulated in CSCs. Tight junction proteins were significantly inhibited by cyclopamine (2, 5μM). Although CD-44, Oct-4 and Nanog were inhibited in CSCs after administration of cyclopamine, these alterations were statistically significant in different genes respectively, but EpCAM was not inhibited. CONCLUSION: EMT, TJ and CSCs markers were affected by Shh signaling pathway in CSCs. Shh signaling pathway may play in an important role of regulation of CSCs.

Cancer stem cells (CSCs) play an important role in cancer development, its main functions are self-renewing capacity, chemoresistance and tumorigeniccapacity. The aim of this study is to clarify the possible role of Shh signaling in regulation of CSCs.METHODS:Normal cancer cells (HCT-116) were cultured with serum medium and cancer stem-like cells (CSCs) were obtained from serum-free medium after incubation for14 days. After cell culturing was done RNA extraction and cDNA transcription of NCs and CSCs (HCT-116). The expressions mRNA of surface markers (CD44,EpCAM), stemness genes (Oct-4, Nanog), Shh signaling (Ptch1, SMO), and shh pathway downstream gene (Gli1), EMT markers (E-Cadherin, Vimentin) and TJgenes (Claudin-4, Occludin) were determined by real time RT-PCR before and after administration of cyclopamine (2, 5 μM).RESULTS:The expressions of surface markers (CD44, EpCAM) and stemness genes (Oct-4, Nanog) were significantly highly expressed in CSCs. Shh signaling pathwayPtch1, SMO and downstream gene Gli1 were significantly higher in CSCs than in NCs. Epithelial marker E-Cadherin was reduced in CSCs, mesenchymal markerVimentin was up-regulated in CSCs. The expressions of Claudin-4 and Occludin were significantly higher in CSCs compared with NCs. SMO, Gli1 and Vimnetin were significantly inhibited after administration of cyclopamine (2, 5μM), but E-Cadherin was up-regulated in CSCs. Tight junction proteins were significantly inhibited by cyclopamine (2, 5μM). Although CD-44, Oct-4 and Nanog were inhibited in CSCs after administration of cyclopamine, these alterations were statistically significant in different genes respectively, but EpCAM was not inhibited.CONCLUSION:EMT, TJ and CSCs markers were affected by Shh signaling pathway in CSCs. Shh signaling pathway may play in an important role of regulation of CSCs.

Background: Laparoscopic gastrectomy has several difficult points including, lymph node dissection and resection of several blood vessels for trainee surgeons. Recently, preoperative evaluation of vasculature based three-dimensional (3D) imaging technique resulted in a significantly improved clinical outcome in abdominal surgery. The aim of this study is to investigate the usefulness of the 3D image in laparoscopic gastrectomy for trainee surgeons. Method: We adopted a multiphase CT protocol to acquire 3 image sets (arterial, portal, and equilibrium phases). 3D-reconstruction of gastric vasculature was made using data from a contrast enhanced MDCT and SYNAPSE VINCENT software. Whole pancreas, spleen, gastric vasculature were extracted from MDCT scans and traced. Thirty three patients, who underwent laparoscopic gastrectomy for gastric cancer during the period between Jan 2013 and May 2014 were examined in this study. Four trainees performed a 19 laparoscopic gastrectomy, while 14 laparoscopic gastrectomy were conducted by the two trainers. The surgical outcomes in both groups and the pattern of gastric vasculatures were evaluated. Result: 3D imaging technique showed a correct positional relationship between the stomach, gastric vessels, pancreas and spleen. Surgical outcome including estimated blood loss, and operative time in trainee group were not significantly different compared to trainer group. 3D imaging technique showed a correct positional relationship between the stomach, gastric vessels, pancreas and spleen. Regarding vascular pattern detected by 3D imaging, the origins of IPA were RGEA in 12 cases (36%), GDA in 8 cases (24%).bifurcation of RGEA and GDA in7 cases (21%), and not detected in 1 case (3%), respectively. The types of confluence of IPV were RGEV in 16 cases (48%), ASPDV in 10 cases (30%), and not detected in 7 cases (21%), respectively. Conclusions: 3D imaging technique might contribute to successful laparoscopic gastrectomy. Preoperative 3D-simulation techniques enabled trainee surgeons to easily and safely perform laparoscopic gastrectomy.

Background: Laparoscopic gastrectomy has several difficult points including,lymph node dissection and resection of several blood vessels for trainee surgeons.Recently, preoperative evaluation of vasculature based three-dimensional (3D)imaging technique resulted in a significantly improved clinical outcome inabdominal surgery. The aim of this study is to investigate the usefulness of the 3Dimage in laparoscopic gastrectomy for trainee surgeons.Method: We adopted a multiphase CT protocol to acquire 3 image sets (arterial,portal, and equilibrium phases). 3D-reconstruction of gastric vasculature wasmade using data from a contrast enhanced MDCT and SYNAPSE VINCENTsoftware. Whole pancreas, spleen, gastric vasculature were extracted from MDCTscans and traced. Thirty three patients, who underwent laparoscopic gastrectomyfor gastric cancer during the period between Jan 2013 and May 2014 wereexamined in this study. Four trainees performed a 19 laparoscopic gastrectomy,while 14 laparoscopic gastrectomy were conducted by the two trainers. Thesurgical outcomes in both groups and the pattern of gastric vasculatures wereevaluated.Result: 3D imaging technique showed a correct positional relationship betweenthe stomach, gastric vessels, pancreas and spleen. Surgical outcome includingestimated blood loss, and operative time in trainee group were not significantlydifferent compared to trainer group. 3D imaging technique showed a correctpositional relationship between the stomach, gastric vessels, pancreas and spleen.Regarding vascular pattern detected by 3D imaging, the origins of IPA were RGEAin 12 cases (36%), GDA in 8 cases (24%).bifurcation of RGEA and GDA in7 cases(21%), and not detected in 1 case (3%), respectively. The types of confluence ofIPV were RGEV in 16 cases (48%), ASPDV in 10 cases (30%), and not detectedin 7 cases (21%), respectively.Conclusions: 3D imaging technique might contribute to successful laparoscopicgastrectomy. Preoperative 3D-simulation techniques enabled trainee surgeons toeasily and safely perform laparoscopic gastrectomy.