目的探究负载膜联蛋白A2(ANXA2)的骨髓间充质干细胞(BMSC)来源的外泌体(Exo-ANXA2)对前列腺癌细胞增殖、侵袭与迁移以及前列腺癌移植瘤生长的影响,并研究巨噬细胞是否参与该过程。方法分离与培养BALB/c裸鼠BMSC,采用负载ANXA2的慢病毒...目的探究负载膜联蛋白A2(ANXA2)的骨髓间充质干细胞(BMSC)来源的外泌体(Exo-ANXA2)对前列腺癌细胞增殖、侵袭与迁移以及前列腺癌移植瘤生长的影响,并研究巨噬细胞是否参与该过程。方法分离与培养BALB/c裸鼠BMSC,采用负载ANXA2的慢病毒质粒感染BMSC,并分离外泌体;加入外泌体处理THP-1巨噬细胞,ELISA检测细胞上清培养液肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、IL-6和IL-10的水平;将外泌体处理的巨噬细胞与前列腺癌细胞共培养后,CCK-8法检测细胞增殖活性,Transwell^(TM)小室检测细胞侵袭与迁移。通过注射PC-3人前列腺癌细胞构建前列腺癌裸鼠移植瘤模型,将建模后的裸鼠随机分为对照组和实验组,每组8只,实验组裸鼠尾静脉注射1 mL Exo-ANXA2,对照组注射等量PBS,于注射后第0、3、6、9、12、15、18、21天,使用游标卡尺测量计算肿瘤体积,21 d处死裸鼠并测量肿瘤组织质量,免疫组织化学染色法检测肿瘤组织中抗原KI-67(ki67)和CD163表达。结果骨髓分离的细胞表面CD90和CD44呈高表达,CD34和CD45呈低表达,且成骨成脂分化能力较强,成功获得BMSC;负载ANXA2的慢病毒质粒感染后,BMSC中有较强的绿色荧光蛋白表达,并成功分离到Exo-ANXA2;经Exo-ANXA2处理后,THP-1细胞中TNF-α和IL-6的水平显著升高而IL-10和IL-13的水平显著下降;Exo-ANXA2处理巨噬细胞后,显著抑制Exo-ANXA2促进PC-3细胞增殖活性、侵袭与迁移的作用;经过给予前列腺癌移植瘤裸鼠注射Exo-ANXA2后,裸鼠在第6、9、12、15、18、21天的肿瘤组织体积显著减小,21 d时裸鼠肿瘤体质量也显著减少,此外,肿瘤组织内ki67和CD163的阳性表达率均显著降低。结论Exo-ANXA2能够抑制前列腺癌细胞增殖、侵袭与迁移,并抑制裸鼠前列腺癌移植瘤生长,M2型巨噬细胞减少。展开更多
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 ng/mL) than in the well differentiated group (20.43 ± 4.97 ng/mL), and in the tumor node metastasis stages Ⅲ-Ⅳ(P 〈 0.01) than in stages Ⅰ-Ⅱ. ANXA2 was not correlated with patient sex, age, size or α-fetoprotein (AFP) level. Area under the receiver operating charac- teristic curve for the whole range of sensitivities and specificities was 0.796 for ANXA2 and 0.782 for AFP. Combining detection of serum ANXA2 and AFP substan- tially improved the diagnostic efficiency (96.52%) and the neclative predictive value ('96.61%) for HCC.of ANXA2 expression has good diagnostic potential for HCC diagnosis.展开更多
Hepatocellular carcinoma(HCC) is the most common primary liver cancer with a dismal prognosis, especially when diagnosed at advanced stages. Annexin A2(ANXA2), is found to promote cancer progression and therapeutic re...Hepatocellular carcinoma(HCC) is the most common primary liver cancer with a dismal prognosis, especially when diagnosed at advanced stages. Annexin A2(ANXA2), is found to promote cancer progression and therapeutic resistance.However, the underlining mechanisms of ANXA2 in immune escape of HCC remain poorly understood up to now. Herein, we summarized the molecular function of ANXA2 in HCC and its relationship with prognosis. Furthermore, we tentatively elucidated the underlying mechanism of ANXA2 immune escape of HCC by upregulating the proportion of regulatory T cells and the expression of several inhibitory molecules, and by downregulating the proportion of natural killer cells and dendritic cells and the expression of several inhibitory molecules or effector molecules. We expect a lot of in-depth studies to further reveal the underlying mechanism of ANXA2 in immune escape of HCC in the future.展开更多
AIM To investigate the clinical utility of serum annexin A2(ANXA2) as a diagnostic marker for early hepatocellular carcinoma(HCC).METHODS This study was performed in HCC Clinic of Ain Shams University Hospitals, Cairo...AIM To investigate the clinical utility of serum annexin A2(ANXA2) as a diagnostic marker for early hepatocellular carcinoma(HCC).METHODS This study was performed in HCC Clinic of Ain Shams University Hospitals, Cairo, Egypt and included: Group 1: Fifty patients with early stage HCC(Barcelona Clinic Liver Cancer stage A); Group 2: Twenty five patients with chronic liver disease; and Control Group: Fifteen healthy, age-and sex-matched subjects who were seronegative for viral hepatitis markers. The followinglaboratory investigations were done: Viral hepatitis markers [hepatitis B surface antigen and hepatitis C virus(HCV) antibodies], HCV RNA in HCV antibody-positive patients, serum alpha fetoprotein(AFP), and serum ANXA2 levels.RESULTS In this study, 88% of HCC patients(n = 44) were HCVpositive, while HBV infection represented only 8% of all HCC patients(n = 4); and two patients were negative for both viral markers. A highly significant difference was found between patients with HCC and chronic liver disease as well as controls with regard to serum ANXA2 levels(130, IQR 15-240; 15, IQR 15-17; and 17, IQR 15-30 ng/m L, respectively). The area under the curve of ANXA2 was 0.865; the cut-off value was established to be 18 ng/mL with a diagnostic sensitivity of 74% and a specificity of 88%, while the sensitivity and specificity of AFP at the cut-off value of 200 ng/dL were 20% and 100%, respectively.CONCLUSION Serum ANXA2 may serve as a biomarker for the early detection of HCC.展开更多
文摘目的探究负载膜联蛋白A2(ANXA2)的骨髓间充质干细胞(BMSC)来源的外泌体(Exo-ANXA2)对前列腺癌细胞增殖、侵袭与迁移以及前列腺癌移植瘤生长的影响,并研究巨噬细胞是否参与该过程。方法分离与培养BALB/c裸鼠BMSC,采用负载ANXA2的慢病毒质粒感染BMSC,并分离外泌体;加入外泌体处理THP-1巨噬细胞,ELISA检测细胞上清培养液肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、IL-6和IL-10的水平;将外泌体处理的巨噬细胞与前列腺癌细胞共培养后,CCK-8法检测细胞增殖活性,Transwell^(TM)小室检测细胞侵袭与迁移。通过注射PC-3人前列腺癌细胞构建前列腺癌裸鼠移植瘤模型,将建模后的裸鼠随机分为对照组和实验组,每组8只,实验组裸鼠尾静脉注射1 mL Exo-ANXA2,对照组注射等量PBS,于注射后第0、3、6、9、12、15、18、21天,使用游标卡尺测量计算肿瘤体积,21 d处死裸鼠并测量肿瘤组织质量,免疫组织化学染色法检测肿瘤组织中抗原KI-67(ki67)和CD163表达。结果骨髓分离的细胞表面CD90和CD44呈高表达,CD34和CD45呈低表达,且成骨成脂分化能力较强,成功获得BMSC;负载ANXA2的慢病毒质粒感染后,BMSC中有较强的绿色荧光蛋白表达,并成功分离到Exo-ANXA2;经Exo-ANXA2处理后,THP-1细胞中TNF-α和IL-6的水平显著升高而IL-10和IL-13的水平显著下降;Exo-ANXA2处理巨噬细胞后,显著抑制Exo-ANXA2促进PC-3细胞增殖活性、侵袭与迁移的作用;经过给予前列腺癌移植瘤裸鼠注射Exo-ANXA2后,裸鼠在第6、9、12、15、18、21天的肿瘤组织体积显著减小,21 d时裸鼠肿瘤体质量也显著减少,此外,肿瘤组织内ki67和CD163的阳性表达率均显著降低。结论Exo-ANXA2能够抑制前列腺癌细胞增殖、侵袭与迁移,并抑制裸鼠前列腺癌移植瘤生长,M2型巨噬细胞减少。
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 ng/mL) than in the well differentiated group (20.43 ± 4.97 ng/mL), and in the tumor node metastasis stages Ⅲ-Ⅳ(P 〈 0.01) than in stages Ⅰ-Ⅱ. ANXA2 was not correlated with patient sex, age, size or α-fetoprotein (AFP) level. Area under the receiver operating charac- teristic curve for the whole range of sensitivities and specificities was 0.796 for ANXA2 and 0.782 for AFP. Combining detection of serum ANXA2 and AFP substan- tially improved the diagnostic efficiency (96.52%) and the neclative predictive value ('96.61%) for HCC.of ANXA2 expression has good diagnostic potential for HCC diagnosis.
基金Supported by the National Natural Science Foundation of China,No.81401988China Postdoctoral Science Foundation,No.2019M661907+1 种基金Jiangsu Postdoctoral Science Foundation,No.2019K159 and No.2019Z153General Project of Jiangsu Provincial Health Committee,No.H2019101.
文摘Hepatocellular carcinoma(HCC) is the most common primary liver cancer with a dismal prognosis, especially when diagnosed at advanced stages. Annexin A2(ANXA2), is found to promote cancer progression and therapeutic resistance.However, the underlining mechanisms of ANXA2 in immune escape of HCC remain poorly understood up to now. Herein, we summarized the molecular function of ANXA2 in HCC and its relationship with prognosis. Furthermore, we tentatively elucidated the underlying mechanism of ANXA2 immune escape of HCC by upregulating the proportion of regulatory T cells and the expression of several inhibitory molecules, and by downregulating the proportion of natural killer cells and dendritic cells and the expression of several inhibitory molecules or effector molecules. We expect a lot of in-depth studies to further reveal the underlying mechanism of ANXA2 in immune escape of HCC in the future.
文摘AIM To investigate the clinical utility of serum annexin A2(ANXA2) as a diagnostic marker for early hepatocellular carcinoma(HCC).METHODS This study was performed in HCC Clinic of Ain Shams University Hospitals, Cairo, Egypt and included: Group 1: Fifty patients with early stage HCC(Barcelona Clinic Liver Cancer stage A); Group 2: Twenty five patients with chronic liver disease; and Control Group: Fifteen healthy, age-and sex-matched subjects who were seronegative for viral hepatitis markers. The followinglaboratory investigations were done: Viral hepatitis markers [hepatitis B surface antigen and hepatitis C virus(HCV) antibodies], HCV RNA in HCV antibody-positive patients, serum alpha fetoprotein(AFP), and serum ANXA2 levels.RESULTS In this study, 88% of HCC patients(n = 44) were HCVpositive, while HBV infection represented only 8% of all HCC patients(n = 4); and two patients were negative for both viral markers. A highly significant difference was found between patients with HCC and chronic liver disease as well as controls with regard to serum ANXA2 levels(130, IQR 15-240; 15, IQR 15-17; and 17, IQR 15-30 ng/m L, respectively). The area under the curve of ANXA2 was 0.865; the cut-off value was established to be 18 ng/mL with a diagnostic sensitivity of 74% and a specificity of 88%, while the sensitivity and specificity of AFP at the cut-off value of 200 ng/dL were 20% and 100%, respectively.CONCLUSION Serum ANXA2 may serve as a biomarker for the early detection of HCC.