AIM: To investigate the effects of Annexin A2 (ANXA2) silencing on invasion, migration, and tumorigenic potential of hepatoma cells. METHODS: Human hepatoma cell lines [HepG2, SMMC-7721, SMMC-7402, and MHCC97-H, a nov...AIM: To investigate the effects of Annexin A2 (ANXA2) silencing on invasion, migration, and tumorigenic potential of hepatoma cells. METHODS: Human hepatoma cell lines [HepG2, SMMC-7721, SMMC-7402, and MHCC97-H, a novel human hepatocellular carcinoma (HCC) cell line with high metastasis potential] and a normal hepatocyte cell line(LO2) were used in this study. The protein and mRNA expression levels of ANXA2 were analysed by western blotting and real-time polymerase chain reaction, re-spectively. The intracellular distribution profile of ANXA2 expression was determined by immunofluorescence and immunohistochemistry. Short hairpin RNA target-ing ANXA2 was designed and stably transfected into MHCC97-H cells. Cells were cultured for in vitro analy-ses or subcutaneously injected as xenografts in mice for in vivo analyses. Effects of ANXA2 silencing on cell growth were assessed by cell counting kit-8 (CCK-8) as-say (in vitro ) and tumour-growth assay (in vivo ), on cell cycling was assessed by flow cytometry and propidium iodide staining (in vitro ), and on invasion and migration potential were assessed by transwell assay and wound-healing assay, respectively (both in vitro ). RESULTS: The MHCC97-H cells, which are known to have high metastasis potential, showed the highest lev-el of ANXA2 expression among the four HCC cell types examined; compared to the LO2 cells, the MHCC97-H expression level was 8-times higher. The ANXA2 expres-sion was effectively inhibited (about 80%) by ANXA2-specific small hairpin RNA (shRNA). ANXA2 expression in the MHCC97-H cells was mainly localized to the cel-lular membrane and cytoplasm, and some localization was detected in the nucleus. Moreover, the proliferation of MHCC97-H cells was obviously suppressed by shR-NA-mediated ANXA2 silencing in vitro , and the tumour growth inhibition rate was 38.24% in vivo . The per-centage of MHCC97-H cells in S phase dramatically de-creased (to 27.76%) under ANXA2-silenced conditions. Furthermore, ANXA2-silenced MHCC97-H cells showed lower invasiveness (percentage of invading cells de-creased to 52.16%) and suppressed migratory capacity (migration distance decreased to 63.49%). It is also worth noting that shRNA-mediated silencing of ANXA2 in the MHCC97-H cells led to abnormal apoptosis. CONCLUSION: shRNA-mediated silencing of ANXA2suppresses the invasion, migration, and tumorigenic potential of hepatoma cells, and may represent a useful target of future molecular therapies.展开更多
AIM： To assess the diagnostic performance of follow- up liver computed tomography （CT） for the detection of high-risk esophageal varices in patients treated with Io- coregional therapy for hepatocellular carcinoma ...AIM： To assess the diagnostic performance of follow- up liver computed tomography （CT） for the detection of high-risk esophageal varices in patients treated with Io- coregional therapy for hepatocellular carcinoma （HCC）. METHODS： We prospectively enrolled 100 patients with cirrhosis who underwent transcatheter arterial chemoembolization, radiofrequency ablation or both procedures for HCCs. All patients underwent upper endoscopy and subsequently liver CT. Three radiolo- gists independently evaluated the presence of high-riskesophageal varices with transverse images alone and with three orthogonal multiplanar reformation （MPR） images, respectively. With endoscopic grading as the reference standard, diagnostic performance was as- sessed by using receiver operating characteristic （ROC） curve analysis. RESULTS： The diagnostic performances （areas under the ROC curve） of three observers with transverse im- ages alone were 0.947 ± 0.031, 0.969 ± 0.024, and 0.916 ＋ 0.038, respectively. The mean sensitivity, spec- ificity, positive predicative value （PPV）, and negative predicative value （NPV） with transverse images alone were 90.1%, 86.39%, 70.9%, and 95.9%, respectively. The diagnostic performances, mean sensitivity, specific- ity, PPV, and NPV with three orthogonal MPR images （0.965 ± 0.025, 0.959 ± 0.027, 0.938 ± 0.033, 91.4%, 89.5%, 76.3%, and 96.6%, respectively） were not su- perior to corresponding values with transverse images alone （P 〉 0.05）, except for the mean specificity （P = 0.039）. CONCLUSION： Our results showed excellent diagnos- tic performance, sensitivity and NPV to detect high-risk esophageal varices on follow-up liver CT after Iocore- gional therapy for HCC,展开更多
基金Supported by The Society Development of Nantong,No.HS2012034the Jiangsu Health Projects,No.BL2012053 and No.K201102+1 种基金the Priority Academic Program Development of Jiangsuthe International S and T Cooperation Program of China,No.2013DFA32150
文摘AIM: To investigate the effects of Annexin A2 (ANXA2) silencing on invasion, migration, and tumorigenic potential of hepatoma cells. METHODS: Human hepatoma cell lines [HepG2, SMMC-7721, SMMC-7402, and MHCC97-H, a novel human hepatocellular carcinoma (HCC) cell line with high metastasis potential] and a normal hepatocyte cell line(LO2) were used in this study. The protein and mRNA expression levels of ANXA2 were analysed by western blotting and real-time polymerase chain reaction, re-spectively. The intracellular distribution profile of ANXA2 expression was determined by immunofluorescence and immunohistochemistry. Short hairpin RNA target-ing ANXA2 was designed and stably transfected into MHCC97-H cells. Cells were cultured for in vitro analy-ses or subcutaneously injected as xenografts in mice for in vivo analyses. Effects of ANXA2 silencing on cell growth were assessed by cell counting kit-8 (CCK-8) as-say (in vitro ) and tumour-growth assay (in vivo ), on cell cycling was assessed by flow cytometry and propidium iodide staining (in vitro ), and on invasion and migration potential were assessed by transwell assay and wound-healing assay, respectively (both in vitro ). RESULTS: The MHCC97-H cells, which are known to have high metastasis potential, showed the highest lev-el of ANXA2 expression among the four HCC cell types examined; compared to the LO2 cells, the MHCC97-H expression level was 8-times higher. The ANXA2 expres-sion was effectively inhibited (about 80%) by ANXA2-specific small hairpin RNA (shRNA). ANXA2 expression in the MHCC97-H cells was mainly localized to the cel-lular membrane and cytoplasm, and some localization was detected in the nucleus. Moreover, the proliferation of MHCC97-H cells was obviously suppressed by shR-NA-mediated ANXA2 silencing in vitro , and the tumour growth inhibition rate was 38.24% in vivo . The per-centage of MHCC97-H cells in S phase dramatically de-creased (to 27.76%) under ANXA2-silenced conditions. Furthermore, ANXA2-silenced MHCC97-H cells showed lower invasiveness (percentage of invading cells de-creased to 52.16%) and suppressed migratory capacity (migration distance decreased to 63.49%). It is also worth noting that shRNA-mediated silencing of ANXA2 in the MHCC97-H cells led to abnormal apoptosis. CONCLUSION: shRNA-mediated silencing of ANXA2suppresses the invasion, migration, and tumorigenic potential of hepatoma cells, and may represent a useful target of future molecular therapies.
基金Supported by Grant from the Samsung Medical Center Clinical Research Development Program,No. CRS108-12-1
文摘AIM： To assess the diagnostic performance of follow- up liver computed tomography （CT） for the detection of high-risk esophageal varices in patients treated with Io- coregional therapy for hepatocellular carcinoma （HCC）. METHODS： We prospectively enrolled 100 patients with cirrhosis who underwent transcatheter arterial chemoembolization, radiofrequency ablation or both procedures for HCCs. All patients underwent upper endoscopy and subsequently liver CT. Three radiolo- gists independently evaluated the presence of high-riskesophageal varices with transverse images alone and with three orthogonal multiplanar reformation （MPR） images, respectively. With endoscopic grading as the reference standard, diagnostic performance was as- sessed by using receiver operating characteristic （ROC） curve analysis. RESULTS： The diagnostic performances （areas under the ROC curve） of three observers with transverse im- ages alone were 0.947 ± 0.031, 0.969 ± 0.024, and 0.916 ＋ 0.038, respectively. The mean sensitivity, spec- ificity, positive predicative value （PPV）, and negative predicative value （NPV） with transverse images alone were 90.1%, 86.39%, 70.9%, and 95.9%, respectively. The diagnostic performances, mean sensitivity, specific- ity, PPV, and NPV with three orthogonal MPR images （0.965 ± 0.025, 0.959 ± 0.027, 0.938 ± 0.033, 91.4%, 89.5%, 76.3%, and 96.6%, respectively） were not su- perior to corresponding values with transverse images alone （P 〉 0.05）, except for the mean specificity （P = 0.039）. CONCLUSION： Our results showed excellent diagnos- tic performance, sensitivity and NPV to detect high-risk esophageal varices on follow-up liver CT after Iocore- gional therapy for HCC,
