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annexin A2 silencing inhibits invasion, migration, and tumorigenic potential of hepatoma cells

World Journal of Gastroenterology 2013年第24期19卷 3792-3801页

作者： Hai-Jian Zhang Deng-Fu Yao Min Yao Hua Huang Li Wang Mei-Juan Yan Xiao-Di Yan Xing Gu Wei Wu Shao-Lin Lu Research Centre of Clinical Medicine Affiliated Hospital of Nantong University Department of Pathology Affiliated Hospital of Nantong University Department of Clinical Oncology Affili-ated Hospital of Nantong University

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-s

关键词： annexin A2 Small hairpin RNA Hepatocel-lular carcinoma Invasion Migration Tumorigenic po-tential

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Cotton AnnGh3 Encoding an annexin Protein is Preferentially Expressed in Fibers and Promotes Initiation and Elongation of Leaf Trichomes in Transgenic Arabidopsis

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Journal of Integrative Plant Biology 2013年第10期55卷 902-916页

作者： Bing Li Deng-Di Li Jie Zhang Hui Xia Xiu-Lan Wang Ying Li Xue-Bao Li Hubei Key Laboratory of Genetic Regulation and Integrative Biology College of Life Sciences Central China Normal University

The annexins are a multifamily of calcium-regulated phospholipid-binding proteins. To investigate the roles of annexins in fiber development, four genes encoding putative annexin proteins were isolated from cotton （Gossypium hirsutum） and designated AnnGh3, AnnGh4, AnnGh5, and AnnGh6. Quantitative reverse transcription-polymerase chain reaction （qRT-PCR） results indicated that AnnGh3, AnnGh4, and AnnGh5 were preferentially expressed in fibers, while the transcripts of AnnGh6 were predominantly accumulated in roots. During fiber development, the transcripts of AnnGh3/4/5 genes were mainly accumulated in rapidly elongating fibers. With fiber cells further developed, their expression activity was dramatically declined to a relatively low level. In situ hybridization results indicated that AnnGh3 and AnnGh5were expressed in initiating fiber cells （0-2 DPA）. Additionally, their expression in fibers was also regulated by phytohormones and [Ca2~]. Subcellular localization analysis discovered that AnnGh3 protein was localized in the cytoplasm. Overexpression of AnnGh3 in Arabidopsis resulted in a significant increase in trichome density and length on leaves of the transgenic plants, suggesting that AnnGh3 may be involved in fiber cell initiation and elongation of cotton.

关键词： annexin cotton (Gossypium hirsutum) fiber development gene expression.

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annexin A2 silencing enhances apoptosis of human umbilical vein endothelial cells in vitro

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Asian Pacific Journal of Tropical Medicine 2015年第11期8卷 929-933页

作者： Shu-Le Jiang Dong-Yan Pan Chao Gu Hai-Feng Qin Shi-Hong Zhao Department of Ophthalmology Affiliated Changhai Hospital of Second Military Medical University

Objective:To study the effects of inhibited annexin A2(ANXA2) on human umbilical vein endothelial cells(HUVECs) in ***:Short hairpin RNA(shRNA) targeting ANXA2 was designed and cloned into double marked lentvirial vector GV248 for RNAi to generate the recombinant expression plasmids,which were stably transfected into *** protein and mRNA expression levels of ANXA2 were analyzed by western blotting and realtime polymerase chain reaction,*** proliferation(cell counting kit-8 assay),apoptosis(flow cytometry analysis),the expression(western blotting) and the activity of easpases(enzyme-linked immunosorbent assay) were used to assess the effects of silencing ANXA2 on HUVECs in ***:The plasmids to express ANXA2-specific shRNA were constructed and were infected into HUVEC resulting in the stably transfected experimental(ANXA2-shRNA),control(eontrol-shRNA) and mock(no plasmid) cell lines,which were verified with western blot and real-time ***/ANXA2-shRNA showed an inhibition rate 91.89%of ANXA2 expression compared to the mock ***2 silencing cell strain obviously presented a lower cell proliferation activity compared to the control and mock HUVECs,with an inhibition rate 82.35%on day 7 in *** analysis indicated that the HUVEC/ANXA2-shRNA cells undergoing apoptosis increased by 102.61%compared to the mock HUVECs(P<0.01).Moreover,the activity levels of caspase-3,caspase-8 and caspase-9in HUVEC/ANXA2-shRNA cells were increased and the activated cleaved caspase-3,cleaved Caspase-8 and cleaved caspase-9 were upregulated evidently compared with that of the control and mock HUVECs by 56.29%,89.59%and 144.58%(P<0.01).Conclusions:shRNAmediated silencing of ANX A2 could not only be able to suppress HUVECs:proliferation but to upregulate the enzyme activity of easpases,which bring to an increase of cell apoptosis This work suggested that ANX A2 may represent a useful target of future molecular therapies.

关键词： Retinal neovascularization annexin A2 RNA interfer

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annexin A1: A new immunohistological marker of cholangiocarcinoma

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World Journal of Gastroenterology 2013年第16期19卷 2456-2465页

作者： Nuttanan Hongsrichan Rucksak Rucksaken Yaovalux Chamgramol Porntip Pinlaor Anchalee Techasen Puangrat Yongvanit Narong Khuntikeo Chawalit Pairojkul Somchai Pinlaorr Department of Parasitology Liver Fluke and Cholangiocarcinoma Research Center Faculty of Medicine Khon Kaen University Department of Pathology Liver Fluke and Cholangiocarcinoma Research Center Faculty of Medicine Khon Kaen University Centre for Research and Development in Medical Diagnostic Laboratory Faculty of Associated Medical Sciences Liver Fluke and Cholangiocarcinoma Research Center Faculty of Medicine Khon Kaen University Department of Biochemistry Liver Fluke and Cholangiocarcinoma Research Center Faculty of Medicine Khon Kaen University Department of Surgery Liver Fluke and Cholangiocarcinoma Research Center Faculty of Medicine Khon Kaen University

AIM: To evaluate a new immunohistological marker, annexin A1 (ANXA1), in cholangiocarcinoma (CCA) and hepatocellular carcinoma (HCC). METHODS: Expression of ANXA1 protein was investigated in liver tissues from patients with CCA and HCC by immunohistochemistry. Its expression on differences stages of tumor development was investigated in hamster CCA tissues induced by Opisthorchis viverrini and N -nitrosodimethylamine. Moreover, mRNA expression of ANXA1 was assessed in CCA cell lines by quantitative real-time polymerase chain reaction and silencing of ANXA1 gene expression using small interfering RNA. RESULTS: In human CCA tissue arrays, immunohistochemical analysis revealed that the positive expression of ANXA1 was 94.1% (64/68 cases) consisting of a high expression (66.2%, 45/68 cases) and a low expression (33.8%, 23/68 cases). However, expression of ANXA1 protein was negative in all histologic patterns for HCC (46/46 cases) and healthy individuals (6/6 cases). In hamster with opisthorchiasis-associated CCA, the expression of ANXA1 was observed in the cytoplasm of inflammatory cells, bile duct epithelia and tumor cells. Grading scores of ANXA1 expression were significantly increased with tumor progression. In addition, mRNA expression of ANXA1 significantly increased in all of the various CCA cell lines tested compared to an immortalized human cholangiocyte cell line (MMNK1). Suppressing the ANXA1 gene significantly reduced the matrix metalloproteinase (MMP) 2 and MMP9, and transforming growth factor-β genes, but increased nuclear factor-kB gene expression. CONCLUSION: ANXA1 is highly expressed in CCA, but low in HCC, suggesting it may serve as a new immunohistochemical marker of CCA. ANXA1 may play a role in opisthorchiasis-associated cholangiocarcinogenesis.

关键词： Cholangiocarcinoma Opisthorchis viverrini Hepatocellular carcinoma annexin A1 Biomarker

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annexin A10 expression in colorectal cancers with emphasis on the serrated neoplasia pathway

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World Journal of Gastroenterology 2015年第33期21卷 9749-9757页

作者： Jeong Mo Bae Jung Ho Kim Ye-Young Rhee Nam-Yun Cho Tae-You Kim Gyeong Hoon Kang Laboratory of Epigenetics Cancer Research InstituteSeoul National University College of Medicine Department of Pathology Seoul National University College of Medicine Department of Internal Medicine Seoul National University College of Medicine

AIM: To validate the utility of annexin A10 as a surrogate marker of the serrated neoplasia pathway in invasive colorectal cancers(CRCs).METHODS: A total of 1133 primary CRC patients who underwent surgical resection at Seoul National University Hospital between January 2004 and December 2007 were *** of annexin A10 was evaluated by immunohistochemistry using tissue microarray and paired to our findings on clinicopathologic and molecular characteristics of each *** G island methylator phenotype was determined by Methy Light assay and microsatellite instability was determined by high performance liquid *** and BRAF mutation status was evaluated by direct sequencing and allele-specific *** and stage-specific survival analyses were performed to reveal the prognostic value of annexin A10 ***: annexin A10 expression was observed in 66(5.8%) of the 1133 *** A10 expression was more commonly found in females and was associated with proximal location,ulcerative gross type,advanced T category,N category and TNM *** with annexin A10 expression showed an absence of luminal necrosis,luminal serration and mucin *** with annexin A10 expression were associated with Cp G island methylator phenotype,microsatellite instability and BRAF *** survival analysis,annexin A10 expression was associated with poor overall survival and progression-free survival,especially in stage Ⅳ ***: annexin A10 expression is associated with poor clinical behavior and can be used a supportive surrogate marker of the serrated neoplasia pathway in invasive CRCs.

关键词： annexin A10 Serrated neoplasia pathway CpG island

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annexin A1在紫杉醇耐药卵巢癌中的表达及其临床意义

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中南大学学报（医学版） 2014年第8期39卷 784-789页

作者：邹玲芝曹兰琴易红中南大学湘雅医院妇产科长沙410008 中南大学湘雅医院卫生部肿瘤蛋白质组学重点实验室长沙410008

目的:探讨annexin A1(ANXA1)在卵巢癌紫杉醇耐药细胞及组织中的表达,分析其与卵巢癌紫杉醇耐药及临床病理特征之间的关系。方法:采用Western印迹及实时PCR方法检测卵巢癌紫杉醇耐药细胞株(SKOV3/Taxol-25)及敏感细胞株(SKOV3)中ANXA1的... 详细信息

目的:探讨annexin A1(ANXA1)在卵巢癌紫杉醇耐药细胞及组织中的表达,分析其与卵巢癌紫杉醇耐药及临床病理特征之间的关系。方法:采用Western印迹及实时PCR方法检测卵巢癌紫杉醇耐药细胞株(SKOV3/Taxol-25)及敏感细胞株(SKOV3)中ANXA1的表达差异;采用免疫组织化学SP法检测42例卵巢癌标本中ANXA1蛋白的表达情况,结合临床资料分析ANXA1表达水平与卵巢癌紫杉醇耐药及临床病理特征的关系。结果:耐药细胞株中ANXA1的表达明显低于敏感细胞株(P0.05),而与临床分期相关(P<0.05)。结论:ANXA1在卵巢癌紫杉醇耐药细胞及组织中低表达,提示通过检测其表达情况可预测卵巢癌患者对紫杉醇化学治疗的敏感性。

关键词：卵巢癌 annexin A1 紫杉醇耐药

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annexin A11 G38R及D40G突变通过降低蛋白稳定性引起肌萎缩侧索硬化症的发生

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中南大学学报（医学版） 2018年第6期43卷 577-582页

作者：廖迪廖巧黄操毕方方中南大学湘雅医院神经内科长沙410008 托马斯杰佛逊大学解剖学系美国宾夕法尼亚州费城19107

目的:探讨annexin A11(ANXA11)G38R及D40G突变引起肌萎缩侧索硬化症(amyotrophic lateral sclerosis,ALS)的发病机制。方法:构建编码ANXA11 G38R,ANXA11 D40G突变蛋白及ANXA11野生型蛋白真核表达载体p ANXA11-G38R-Flag,p ANXA11-D40G-Flag和p ANXA11-Flag;将其转染至HEK293细胞后,用放线菌酮分别处理0,2,4,8 h;收集蛋白采用Western印迹检测3种蛋白的含量变化情况,比较目标蛋白的半衰期;构建稳定表达ANXA11 G38R,ANXA11D40G突变蛋白及ANXA11野生型蛋白的NSC-34细胞系,用Western印迹检测上述蛋白在NP-40裂解液中的溶解情况。结果:ANXA11 G38R,ANXA11 D40G突变蛋白的半衰期较野生型蛋白缩短(P0.05);ANXA11 G38R,ANXA11 D40G突变蛋白与ANXA11野生型蛋白在NP-40裂解液中均未发现不可溶成分。结论:G38R和D40G突变降低了ANXA11蛋白的稳定性;G38R和D40G突变不影响ANXA11蛋白在NP-40裂解液中的可溶性。

关键词： annexin A11 肌萎缩侧索硬化症半衰期可溶性

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正电子核素标记annexin Ⅴ的研究进展

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原子能科学技术 2011年第8期45卷 908-914页

作者：郭飞虎陈大明刘卓温凯杜进中国原子能科学研究院原子高科股份有限公司北京102413 中国同位素公司北京100045

annexinⅤ是一种磷脂结合蛋白,能与细胞凋亡过程中翻转到膜外的磷脂酰丝氨酸(PS)特异性结合,因此annexinⅤ是检测细胞早期凋亡的灵敏指针。利用正电子核素对annexinⅤ进行标记后,通过与PS的结合可检测细胞凋亡,实现活体无创正电子发射... 详细信息

annexinⅤ是一种磷脂结合蛋白,能与细胞凋亡过程中翻转到膜外的磷脂酰丝氨酸(PS)特异性结合,因此annexinⅤ是检测细胞早期凋亡的灵敏指针。利用正电子核素对annexinⅤ进行标记后,通过与PS的结合可检测细胞凋亡,实现活体无创正电子发射计算机断层显像(PET),从而进行人体细胞凋亡研究,达到疾病早期诊断的目的。本文就正电子核素标记annexinⅤ用于凋亡显像的研究现状进行综述。

关键词：细胞凋亡 annexin Ⅴ 正电子发射计算机断层显像

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annexin a2表达对人乳腺癌细胞增殖迁移和侵袭能力的影响

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中国肿瘤临床 2011年第6期38卷 304-307页

作者：曹淑贞张飞赵培起韩敬华武冰张海嫦张霖牛瑞芳乳腺癌防治教育部重点实验室天津医科大学附属肿瘤医院肿瘤研究所天津市300060

目的:研究annexin a2的表达对人乳腺癌细胞MDA-MB-231增殖、迁移和侵袭能力的影响。方法:采用小干扰RNA技术下调人乳腺癌细胞中annexin a2的表达,并筛选单克隆细胞株,得到实验组克隆,阴性对照组命名为control。采用Westermblot(WB)技术... 详细信息

目的:研究annexin a2的表达对人乳腺癌细胞MDA-MB-231增殖、迁移和侵袭能力的影响。方法:采用小干扰RNA技术下调人乳腺癌细胞中annexin a2的表达,并筛选单克隆细胞株,得到实验组克隆,阴性对照组命名为control。采用Westermblot(WB)技术检测annexin a2蛋白水平的变化,噻唑蓝(MTT)比色法研究其对MDA-MB-231细胞的存活、增殖能力的影响;采用划痕实验、侵袭实验观察其对细胞迁移和侵袭能力的影响;免疫沉淀法检测与annexin a2相互作用的蛋白分子。结果:WB检测发现siRNA干扰后MDA-MB-231细胞中annexin a2的蛋白表达水平明显下降,MTT增殖实验显示annexin a2下降组细胞克隆在72、96h的增殖能力明显低于亲本和control组细胞(P均annexin a2与热休克蛋白27(HSP27)之间存在相互作用。结论:人乳腺癌细胞MDA-MB-231具有较强的迁移和侵袭能力,annexin a2蛋白水平下降可以明显抑制其增殖、迁移和侵袭能力,研究提示annexin a2与HSP27的相互作用有可能在乳腺癌的发生、浸润和转移中起调节作用。

关键词： annexin a2 乳腺癌增殖迁移侵袭

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annexinⅠ对胰腺癌细胞生物学行为影响的体内研究

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中华肿瘤杂志 2008年第12期30卷 897-900页

作者：刘骞胡海冉宇靓王成锋赵东兵田艳涛孙立新谢亦斌杨治华赵平中国医学科学院肿瘤医院肿瘤研究所腹部外科北京100021 中国医学科学院肿瘤医院细胞生物研究室北京100021

目的探讨annexinⅠ在胰腺癌发生过程中的作用。方法应用RNA干扰技术，敲低annexinⅠ基因在mRNA水平的表达，并经Western blot检测证实。分别接种人胰腺癌Suit-Ⅱ细胞及转染annexinⅠ—siRNA2、annexinⅠ—siRNA3、annexinⅠ—siRNAN的Su... 详细信息

目的探讨annexinⅠ在胰腺癌发生过程中的作用。方法应用RNA干扰技术，敲低annexinⅠ基因在mRNA水平的表达，并经Western blot检测证实。分别接种人胰腺癌Suit-Ⅱ细胞及转染annexinⅠ—siRNA2、annexinⅠ—siRNA3、annexinⅠ—siRNAN的Suit-Ⅱ细胞，建立裸鼠胰腺癌移植瘤模型，观察胰腺癌细胞的成瘤能力、肿瘤生长速度的变化，测定肿瘤体积和瘤重。结果（1）Western blot检测结果显示，转染pSilencer—annexinⅠ—siRNA1的Suit-Ⅱ细胞annexinⅠ表达显著降低，转染pSilencer—annexinⅠ—siRNA2和pSilencer—annexinⅠ—siRNA3的Suit—Ⅱ细胞annexinⅠ的表达几乎被完全抑制。（2）接种转染annexinⅠ—siRNA2、annexinⅠ-siRNA3细胞组的肿瘤生长速度较接种亲本Suit-Ⅱ细胞组明显减慢，肿瘤生长被抑制，抑制率分别达76．6％和68．4％。接种肿瘤细胞后44d，接种转染annexinⅠ-siRNA2细胞组、接种转染annexinⅠ-siRNA3细胞组的瘤重分别为0．8987和0．8992g，显著低于接种亲本Suit-Ⅱ细胞组和接种转染annexinⅠ—siRNAN细胞组（分别为2．5866和2．4070g，P〈0．001）。结论annexinⅠ基因在胰腺癌发生过程中起到了促进胰腺癌细胞生长增殖、增强胰腺癌细胞成瘤能力的重要作用，可作为基因治疗的潜在靶点。

关键词： annexin Ⅰ 胰腺肿瘤 RNA干扰肿瘤生长

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