Journal: Journal of cellular and molecular medicine

Article Title: Secretion of annexin A3 from ovarian cancer cells and its association with platinum resistance in ovarian cancer patients.

doi: 10.1111/j.1582-4934.2011.01316.x

Figure Lengend Snippet: Fig. 1 (A) Annexin A3 levels in the conditional culture medium from the ovarian cancer cells were measured by ELISA. (B) Proteins from the ovarian cancer cell lysates and concentrated cul- ture media were analysed by anti-annexin A3 immunoblotting. Enforced expression of annexin A3 in SKOV3 and A2780 cells resulted in the increased secretion of annexin A3 in the culture medium. Down-regulation of annexin A3 in plat- inum-resistant ovarian cancer cells SKOV3/Cis andA2780/Cis, which express high levels of annexin A3, reduced the secretion of annexin A3. (C) Annexin A3 levels in sera from the 30 normal female donors and the 50 ovarian cancer patients were determined by ELISA [particular high levels of annexin A3 (2.0461, 3.4453, 8.8125 and 18.3081 ng/ml, respectively) cannot be seen in the graph because the Y-axis range is 2.0 ng/ml]. The patients are divided into platinum-sensitive (n 20) and resistant (n 30) groups based on clinical data. There are significant differences in the serum levels of annexin A3 among normal donors, the sensitive group and the resistant group. Data were analysed with a Mann–Whitney U-Test. P values were all two-sided. (D) Progress-free times of the 50 ovarian cancer patients. There is a significant difference between the high annexin A3 group (serum A3 1.13 ng/ml) and the low annexin A3 group (serum A3 1.13 ng/ml) (P 0.009 0.05).

Article Snippet: The level of annexin A3 in sera of ovarian cancer patients was determined using the ELISA kit for human annexin A3 (Cusabio) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, MANN-WHITNEY

Journal: Journal of cellular and molecular medicine

Article Title: Secretion of annexin A3 from ovarian cancer cells and its association with platinum resistance in ovarian cancer patients.

doi: 10.1111/j.1582-4934.2011.01316.x

Figure Lengend Snippet: Fig. 2 High levels of annexin A3 are associated with increased numbers of vesicles in the cytoplasm. (A) Representative TEM (10,000) photographs of paired ovarian cancer cell lines SKOV3 and SKOV3/Cis and A2780 and A2780/Cis. The arrows indicate the increased numbers of vesicles in plat- inum-resistant SKOV3/Cis and A2780/Cis cells. (B) Higher magnification (50,000) photographs of SKOV3/Cis cells. Arrows indicate the MVB-like vesicle (upper) and exocytosis of a vesicle (lower). (C) Representative TEM (10,000) photographs of ovarian cancer cell lines. Arrows in the upper panel indicated increased vesicles in annexin A3-expressing A2780/Ann and SKOV3/Ann cells. The lower panels are pictures of A2780/Cis/R and SKOV3/Cis/R cells. The vesicles observed in A2780/Cis and SKOV3/Cis (A) have largely disappeared.

Article Snippet: The level of annexin A3 in sera of ovarian cancer patients was determined using the ELISA kit for human annexin A3 (Cusabio) according to the manufacturer’s instructions.

Techniques: Expressing

Journal: Journal of cellular and molecular medicine

Article Title: Secretion of annexin A3 from ovarian cancer cells and its association with platinum resistance in ovarian cancer patients.

doi: 10.1111/j.1582-4934.2011.01316.x

Figure Lengend Snippet: Fig. 3 Immunoelectron microscopy studies of annexin A3 in platinum-resistant ovarian cancer cell SKOV3/Cis. Cell preparation and treatment are described in Materials and Methods. Ultrathin sections were examined by transmission elec- tron microscopy (left, 50,000; right, 100,000). The high-density 12-nm colloidal gold particles, which are indicative of annexin A3, could be observed in the cytoplasm of the cells (small arrows). Some gold particles existed in or around vesicles (medium arrows). (A) Control (sections were treated as described without anti- annexin A3 antibody). (B) Annexin A3 expression in cytoplasm. (C, D) Annexin A3 expression in cytoplamic vesicles. The vesicle in C appears ready to undergo exocytosis.

Article Snippet: The level of annexin A3 in sera of ovarian cancer patients was determined using the ELISA kit for human annexin A3 (Cusabio) according to the manufacturer’s instructions.

Techniques: Immuno-Electron Microscopy, Transmission Assay, Microscopy, Control, Expressing

Journal: Journal of cellular and molecular medicine

Article Title: Secretion of annexin A3 from ovarian cancer cells and its association with platinum resistance in ovarian cancer patients.

doi: 10.1111/j.1582-4934.2011.01316.x

Figure Lengend Snippet: Fig. 4 Annexin A3 is associated with exosomes from ovarian cancer cells SKOV3/Cis. (A) Electron microscopic picture of exosomes from SKOV3/Cis cells. The exosomes are 40–100 nm in diameter and exhibit the typical cup-shaped morphology. (B, C) IEM photographs of exo- somes from SKOV3/Cis cells. The high-density dots represent 12-nm gold particles, which are indicatives of annexin A3. (D) Conditional medium (30 ml) from cultured SKOV3/Cis cells was collected, and divided into two parts. After removing the exosomes from one of them (DMEM 2), both were concentrated and analysed by immunobltting with anti-annexin A3 and anti- Hsp70 antibodies. The exosome preparation and the upper sucrose were also examined by the immunoblotting.

Article Snippet: The level of annexin A3 in sera of ovarian cancer patients was determined using the ELISA kit for human annexin A3 (Cusabio) according to the manufacturer’s instructions.

Techniques: Cell Culture, Western Blot

Journal: Journal of cellular and molecular medicine

Article Title: Secretion of annexin A3 from ovarian cancer cells and its association with platinum resistance in ovarian cancer patients.

doi: 10.1111/j.1582-4934.2011.01316.x

Figure Lengend Snippet: Fig. 5 The amount of exosomes released by the ovarian cancer cells depends on the expression of annexin A3. (A) Exosomes were isolated from condi- tioned medium of cultured ovarian cancer cells. The total amount of protein was used to quantify recovered exosomes. (B) Comparison of the amounts of exosomes released from various ovarian cancer cells by immunobltting with anti-annexin A3 and anti-Hsp70 antibodies.

Article Snippet: The level of annexin A3 in sera of ovarian cancer patients was determined using the ELISA kit for human annexin A3 (Cusabio) according to the manufacturer’s instructions.

Techniques: Expressing, Isolation, Cell Culture, Comparison

Journal: Translational cancer research

Article Title: ANXA3 is upregulated by hypoxia-inducible factor 1-alpha and promotes colon cancer growth.

doi: 10.21037/tcr-20-994

Figure Lengend Snippet: Figure 1 Immunohistochemical staining of HIF-1α and ANXA3 in colon cancer and normal colon tissues. Positive expression was visualized

Article Snippet: Next, the membranes were blocked in 5% non-fat dry milk for 1 h. Then the membranes were probed by antibody for HIF1α (1:1,000), ANXA3 (1:800) or β-actin (1:500, Boster, Biotechnology, Wuhan, China) overnight at 4 °C.

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Translational cancer research

Article Title: ANXA3 is upregulated by hypoxia-inducible factor 1-alpha and promotes colon cancer growth.

doi: 10.21037/tcr-20-994

Figure Lengend Snippet: Figure 3 Detection of HIF-1α and ANXA3 protein levels in SW480, SW620, HCT116 and HT29 colon cancer cells under normoxic

Article Snippet: Next, the membranes were blocked in 5% non-fat dry milk for 1 h. Then the membranes were probed by antibody for HIF1α (1:1,000), ANXA3 (1:800) or β-actin (1:500, Boster, Biotechnology, Wuhan, China) overnight at 4 °C.

Techniques:

Journal: Translational cancer research

Article Title: ANXA3 is upregulated by hypoxia-inducible factor 1-alpha and promotes colon cancer growth.

doi: 10.21037/tcr-20-994

Figure Lengend Snippet: Figure 2 Detection of HIF-1α and ANXA3 expression in colon cancer and normal colon tissues. (A) qRT-PCR analysis of HIF-1α and

Article Snippet: Next, the membranes were blocked in 5% non-fat dry milk for 1 h. Then the membranes were probed by antibody for HIF1α (1:1,000), ANXA3 (1:800) or β-actin (1:500, Boster, Biotechnology, Wuhan, China) overnight at 4 °C.

Techniques: Expressing, Quantitative RT-PCR

Journal: Translational cancer research

Article Title: ANXA3 is upregulated by hypoxia-inducible factor 1-alpha and promotes colon cancer growth.

doi: 10.21037/tcr-20-994

Figure Lengend Snippet: Figure 4 HIF-1α regulates ANXA3 expression in SW480 and SW620 cells. The expression of HIF-1α and ANXA3 was measured by qRT-

Article Snippet: Next, the membranes were blocked in 5% non-fat dry milk for 1 h. Then the membranes were probed by antibody for HIF1α (1:1,000), ANXA3 (1:800) or β-actin (1:500, Boster, Biotechnology, Wuhan, China) overnight at 4 °C.

Techniques: Expressing

Journal: Translational cancer research

Article Title: ANXA3 is upregulated by hypoxia-inducible factor 1-alpha and promotes colon cancer growth.

doi: 10.21037/tcr-20-994

Figure Lengend Snippet: Figure 6 HIF-1α knockdown decreases tumor volume and ANXA3 expression in nude xenograft tumor model. (A) Compared with flank

Article Snippet: Next, the membranes were blocked in 5% non-fat dry milk for 1 h. Then the membranes were probed by antibody for HIF1α (1:1,000), ANXA3 (1:800) or β-actin (1:500, Boster, Biotechnology, Wuhan, China) overnight at 4 °C.

Techniques: Knockdown, Expressing

Journal: PLoS ONE

Article Title: Identifying Resistance Mechanisms against Five Tyrosine Kinase Inhibitors Targeting the ERBB/RAS Pathway in 45 Cancer Cell Lines

doi: 10.1371/journal.pone.0059503

Figure Lengend Snippet: Validation of the top genes by TaqMan RT-PCR in the cell lines.

Article Snippet: ANXA3 , Hs00971411_m1 , 209369_at , annexin A3 , gefitinib , 0.040.

Techniques: Biomarker Discovery, Binding Assay

Journal: PLoS ONE

Article Title: Identifying Resistance Mechanisms against Five Tyrosine Kinase Inhibitors Targeting the ERBB/RAS Pathway in 45 Cancer Cell Lines

doi: 10.1371/journal.pone.0059503

Figure Lengend Snippet: Circos plot of RT-PCR validated correlations for genes associated with resistance against multiple agents as identified by microarray analysis. The thickness of the ribbons correlate to the log(p) of the correlation (see .). Note the high number of genes associated with sunitinib resistance and the single gene associated with lapatinib resistance. The two most informative genes are ANXA3 and RAB25, each associated with resistance against four agents.

Article Snippet: ANXA3 , Hs00971411_m1 , 209369_at , annexin A3 , gefitinib , 0.040.

Techniques: Reverse Transcription Polymerase Chain Reaction, Microarray

Journal: Journal of Cellular and Molecular Medicine

Article Title: Secretion of annexin A3 from ovarian cancer cells and its association with platinum resistance in ovarian cancer patients

doi: 10.1111/j.1582-4934.2011.01316.x

Figure Lengend Snippet: Annexin A3 levels in the conditional culture medium from the ovarian cancer cells were measured by ELISA. (B) Proteins from the ovarian cancer cell lysates and concentrated culture media were analysed by anti-annexin A3 immunoblotting. Enforced expression of annexin A3 in SKOV3 and A2780 cells resulted in the increased secretion of annexin A3 in the culture medium. Down-regulation of annexin A3 in platinum-resistant ovarian cancer cells SKOV3/Cis andA2780/Cis, which express high levels of annexin A3, reduced the secretion of annexin A3. (C) Annexin A3 levels in sera from the 30 normal female donors and the 50 ovarian cancer patients were determined by ELISA [particular high levels of annexin A3 (2.0461, 3.4453, 8.8125 and 18.3081 ng/ml, respectively) cannot be seen in the graph because the Y -axis range is 2.0 ng/ml]. The patients are divided into platinum-sensitive ( n = 20) and resistant ( n = 30) groups based on clinical data. There are significant differences in the serum levels of annexin A3 among normal donors, the sensitive group and the resistant group. Data were analysed with a Mann–Whitney U -Test. P values were all two-sided. (D) Progress-free times of the 50 ovarian cancer patients. There is a significant difference between the high annexin A3 group (serum A3 > 1.13 ng/ml) and the low annexin A3 group (serum A3 < 1.13 ng/ml) ( P = 0.009 < 0.05).

Article Snippet: The level of annexin A3 in sera of ovarian cancer patients was determined using the ELISA kit for human annexin A3 (Cusabio) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, MANN-WHITNEY

Journal: Journal of Cellular and Molecular Medicine

Article Title: Secretion of annexin A3 from ovarian cancer cells and its association with platinum resistance in ovarian cancer patients

doi: 10.1111/j.1582-4934.2011.01316.x

Figure Lengend Snippet: Annexin A3 levels in sera from ovarian cancer patients and normal female donors

Article Snippet: The level of annexin A3 in sera of ovarian cancer patients was determined using the ELISA kit for human annexin A3 (Cusabio) according to the manufacturer’s instructions.

Techniques:

Journal: Journal of Cellular and Molecular Medicine

Article Title: Secretion of annexin A3 from ovarian cancer cells and its association with platinum resistance in ovarian cancer patients

doi: 10.1111/j.1582-4934.2011.01316.x

Figure Lengend Snippet: Clinical characteristics of the four patients with high-level serum annexin A3 (> 2 ng/ml)

Article Snippet: The level of annexin A3 in sera of ovarian cancer patients was determined using the ELISA kit for human annexin A3 (Cusabio) according to the manufacturer’s instructions.

Techniques:

Journal: Journal of Cellular and Molecular Medicine

Article Title: Secretion of annexin A3 from ovarian cancer cells and its association with platinum resistance in ovarian cancer patients

doi: 10.1111/j.1582-4934.2011.01316.x

Figure Lengend Snippet: High levels of annexin A3 are associated with increased numbers of vesicles in the cytoplasm. (A) Representative TEM (10,000×) photographs of paired ovarian cancer cell lines SKOV3 and SKOV3/Cis and A2780 and A2780/Cis. The arrows indicate the increased numbers of vesicles in platinum-resistant SKOV3/Cis and A2780/Cis cells. (B) Higher magnification (50,000×) photographs of SKOV3/Cis cells. Arrows indicate the MVB-like vesicle (upper) and exocytosis of a vesicle (lower). (C) Representative TEM (10,000×) photographs of ovarian cancer cell lines. Arrows in the upper panel indicated increased vesicles in annexin A3-expressing A2780/Ann and SKOV3/Ann cells. The lower panels are pictures of A2780/Cis/R and SKOV3/Cis/R cells. The vesicles observed in A2780/Cis and SKOV3/Cis (A) have largely disappeared.

Article Snippet: The level of annexin A3 in sera of ovarian cancer patients was determined using the ELISA kit for human annexin A3 (Cusabio) according to the manufacturer’s instructions.

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Secretion of annexin A3 from ovarian cancer cells and its association with platinum resistance in ovarian cancer patients

doi: 10.1111/j.1582-4934.2011.01316.x

Figure Lengend Snippet: Immunoelectron microscopy studies of annexin A3 in platinum-resistant ovarian cancer cell SKOV3/Cis. Cell preparation and treatment are described in Materials and Methods. Ultrathin sections were examined by transmission electron microscopy (left, 50,000×; right, 100,000×). The high-density 12-nm colloidal gold particles, which are indicative of annexin A3, could be observed in the cytoplasm of the cells (small arrows). Some gold particles existed in or around vesicles (medium arrows). (A) Control (sections were treated as described without anti-annexin A3 antibody). (B) Annexin A3 expression in cytoplasm. (C, D) Annexin A3 expression in cytoplamic vesicles. The vesicle in C appears ready to undergo exocytosis.

Article Snippet: The level of annexin A3 in sera of ovarian cancer patients was determined using the ELISA kit for human annexin A3 (Cusabio) according to the manufacturer’s instructions.

Techniques: Immuno-Electron Microscopy, Transmission Assay, Electron Microscopy, Control, Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Secretion of annexin A3 from ovarian cancer cells and its association with platinum resistance in ovarian cancer patients

doi: 10.1111/j.1582-4934.2011.01316.x

Figure Lengend Snippet: Annexin A3 is associated with exosomes from ovarian cancer cells SKOV3/Cis. (A) Electron microscopic picture of exosomes from SKOV3/Cis cells. The exosomes are 40–100 nm in diameter and exhibit the typical cup-shaped morphology. (B, C) IEM photographs of exosomes from SKOV3/Cis cells. The high-density dots represent 12-nm gold particles, which are indicatives of annexin A3. (D) Conditional medium (30 ml) from cultured SKOV3/Cis cells was collected, and divided into two parts. After removing the exosomes from one of them (DMEM 2), both were concentrated and analysed by immunobltting with anti-annexin A3 and anti-Hsp70 antibodies. The exosome preparation and the upper sucrose were also examined by the immunoblotting.

Article Snippet: The level of annexin A3 in sera of ovarian cancer patients was determined using the ELISA kit for human annexin A3 (Cusabio) according to the manufacturer’s instructions.

Techniques: Cell Culture, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Secretion of annexin A3 from ovarian cancer cells and its association with platinum resistance in ovarian cancer patients

doi: 10.1111/j.1582-4934.2011.01316.x

Figure Lengend Snippet: The amount of exosomes released by the ovarian cancer cells depends on the expression of annexin A3. (A) Exosomes were isolated from conditioned medium of cultured ovarian cancer cells. The total amount of protein was used to quantify recovered exosomes. (B) Comparison of the amounts of exosomes released from various ovarian cancer cells by immunobltting with anti-annexin A3 and anti-Hsp70 antibodies.

Article Snippet: The level of annexin A3 in sera of ovarian cancer patients was determined using the ELISA kit for human annexin A3 (Cusabio) according to the manufacturer’s instructions.

Techniques: Expressing, Isolation, Cell Culture, Comparison