AIM： TO investigate the characteristics and diagnostic value of annexin A2 （ANXA2） expression in cancerous tissues and sera of patients with hepatitis B virus （HBV）-related hepatocellular carcinoma （HCC）. METH...AIM： TO investigate the characteristics and diagnostic value of annexin A2 （ANXA2） expression in cancerous tissues and sera of patients with hepatitis B virus （HBV）-related hepatocellular carcinoma （HCC）. METHODS： Levels of liver ANXA2 gene transcription or protein expression were analyzed in HCC-, their self- controlled precancerous-, and distant cancerous- tissues from 30 HCC. Serum levels of ANXA2 expression in 115 patients with HCC, 25 with metastatic liver can cer, 35 with chronic hepatitis, 28 with acute hepatitis, 38 with cirrhosis, and 30 healthy controls were deter- mined. Clinicopathological characteristics of circulating ANXA2 expression were analyzed, and its diagnostic efficiency and clinical values in HCC were evaluated. RESULTS： ANXA2 expression was localized in both cell membrane and cytoplasm in HCC tissue, mainly in the cytoplasm of matched adjacent cancerous tissue, and there was almost no positive staining in matched distant cancerous tissue. Abnormal expression of liver ANXA2 was present in HCC tissues compared with self-con- trolled adjacent- and distant-cancerous tissues at pro- tein or mRNA level. Circulating ANXA2 in HCC patients was significantly higher than that of other liver diseases （P 〈 0.01） except metastatic liver cancer. If the diag- nostic cutoff value of ANXA2 level was more than 18 ng/ mL, the incidence of serum ANXA2 was 86.96% in the HCC group, 80% in the metastatic liver cancer group, 31.58% in the liver cirrhosis group, none in the chronic hepatitis or acute hepatitis or normal control group, respectively. Serum ANXA2 expression in HCC patients was correlated with HBV infection （27.38 ± 5.67 ng/mL vs 18.58 ± 7.83 ng/mL, P 〈 0.01）, extrahepatic metas- tasis （26.11±5.43 ng/mL ys 22.79 ± 5.64 ng/mL, P 〈 0.01）, and portal vein thrombus （26.03 ± 5.99 ng/mL vs 23.06 ± 5.03 ng/mL, P 〈 0.01）, and was significantly higher （P 〈 0.01） in the moderately- （26.19±5.34 ng/ mL） or the poorly- differentiated group （27.05 ± 5.13 展开更多
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展开更多
基金Supported by Priority Academic Program Development of Jiangsu Higher Education Institution (PAPD)the Project of Jiangsu Clinical Medicine (BL2012053)+1 种基金the Programs of Nantong Society Undertaking and Technological Innovation,No.HS2012034 and HS2011012the International S and T Cooperation Program of China
文摘AIM： TO investigate the characteristics and diagnostic value of annexin A2 （ANXA2） expression in cancerous tissues and sera of patients with hepatitis B virus （HBV）-related hepatocellular carcinoma （HCC）. METHODS： Levels of liver ANXA2 gene transcription or protein expression were analyzed in HCC-, their self- controlled precancerous-, and distant cancerous- tissues from 30 HCC. Serum levels of ANXA2 expression in 115 patients with HCC, 25 with metastatic liver can cer, 35 with chronic hepatitis, 28 with acute hepatitis, 38 with cirrhosis, and 30 healthy controls were deter- mined. Clinicopathological characteristics of circulating ANXA2 expression were analyzed, and its diagnostic efficiency and clinical values in HCC were evaluated. RESULTS： ANXA2 expression was localized in both cell membrane and cytoplasm in HCC tissue, mainly in the cytoplasm of matched adjacent cancerous tissue, and there was almost no positive staining in matched distant cancerous tissue. Abnormal expression of liver ANXA2 was present in HCC tissues compared with self-con- trolled adjacent- and distant-cancerous tissues at pro- tein or mRNA level. Circulating ANXA2 in HCC patients was significantly higher than that of other liver diseases （P 〈 0.01） except metastatic liver cancer. If the diag- nostic cutoff value of ANXA2 level was more than 18 ng/ mL, the incidence of serum ANXA2 was 86.96% in the HCC group, 80% in the metastatic liver cancer group, 31.58% in the liver cirrhosis group, none in the chronic hepatitis or acute hepatitis or normal control group, respectively. Serum ANXA2 expression in HCC patients was correlated with HBV infection （27.38 ± 5.67 ng/mL vs 18.58 ± 7.83 ng/mL, P 〈 0.01）, extrahepatic metas- tasis （26.11±5.43 ng/mL ys 22.79 ± 5.64 ng/mL, P 〈 0.01）, and portal vein thrombus （26.03 ± 5.99 ng/mL vs 23.06 ± 5.03 ng/mL, P 〈 0.01）, and was significantly higher （P 〈 0.01） in the moderately- （26.19±5.34 ng/ mL） or the poorly- differentiated group （27.05 ± 5.13
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
基金Supported by Grants from the State Key Projects for Basic Re-search,No.2011CB910703 and No.2012ZX10002-017(to Zhao XH)National High-tech R and D Program,No.2013AA041201(to Qian YM)and No.2012AA020206(to Zhao XH)+1 种基金National Natural Science Foundation of China,No.81372591 and No.81321091(to Zhao XH)the Research Foundation of the Center for Marine Medicine and Rescue of Tsinghua University and NGH(to Qian YM and Zhao XH)
文摘AIM: To develop a novel method for the rapid and efficient extraction of exosomes secreted by tumor cells.
