Journal: Pharmaceutics

Article Title: Synergistic Encapsulation of Paclitaxel and Sorafenib by Methoxy Poly(Ethylene Glycol)- b -Poly(Caprolactone) Polymeric Micelles for Ovarian Cancer Therapy

doi: 10.3390/pharmaceutics15041206

Figure Lengend Snippet: IC 50 and CI values of paclitaxel (PTX) and sorafenib (SRF) at various molar ratios (n = 3).

Article Snippet: SKOV3-red-fluc, a cancer cell line in which a fluorescent factor was introduced into SKOV3, a human OC cell line, was purchased from PerkinElmer (Waltham, MA, USA).

Techniques:

Journal: Pharmaceutics

Article Title: Synergistic Encapsulation of Paclitaxel and Sorafenib by Methoxy Poly(Ethylene Glycol)- b -Poly(Caprolactone) Polymeric Micelles for Ovarian Cancer Therapy

doi: 10.3390/pharmaceutics15041206

Figure Lengend Snippet: In vitro cytotoxicity analysis of SKOV3-red-fluc ( A ) PTX solution, ( B ) PTX micelle, ( C ) SRF solution, ( D ) SRF micelle, ( E ) PTX/SRF solution, and ( F ) PTX/SRF micelle (n = 6).

Article Snippet: SKOV3-red-fluc, a cancer cell line in which a fluorescent factor was introduced into SKOV3, a human OC cell line, was purchased from PerkinElmer (Waltham, MA, USA).

Techniques: In Vitro

Journal: Pharmaceutics

Article Title: Synergistic Encapsulation of Paclitaxel and Sorafenib by Methoxy Poly(Ethylene Glycol)- b -Poly(Caprolactone) Polymeric Micelles for Ovarian Cancer Therapy

doi: 10.3390/pharmaceutics15041206

Figure Lengend Snippet: Colony formation inhibition in SKOV3-red-fluc cell line solution and micelles ( A ) free PTX, ( B ) PTX micelle, ( C ) free SRF, ( D ) SRF micelle, ( E ) free PTX/SRF, and ( F ) PTX/SRF micelle (n = 3).

Article Snippet: SKOV3-red-fluc, a cancer cell line in which a fluorescent factor was introduced into SKOV3, a human OC cell line, was purchased from PerkinElmer (Waltham, MA, USA).

Techniques: Inhibition

Journal: Pharmaceutics

Article Title: Synergistic Encapsulation of Paclitaxel and Sorafenib by Methoxy Poly(Ethylene Glycol)- b -Poly(Caprolactone) Polymeric Micelles for Ovarian Cancer Therapy

doi: 10.3390/pharmaceutics15041206

Figure Lengend Snippet: ( A ) Morphological observation of tumor spheroids prepared with SKOV3-red-fluc, ( B ) Control, PTX Solution, PTX micelles, PTX/SRF Solution, and PTX/SRF micelles SKOV3-red-fluc tumor spheroid IVIS measured with D-luciferin before treatment and after secondary treatment image (n = 4). ( C ) A graph showing the total luminescence value reflecting the tumor size for the first and second drug treatments based on the tumor spheroid before drug treatment (** p < 0.01, n = 4).

Article Snippet: SKOV3-red-fluc, a cancer cell line in which a fluorescent factor was introduced into SKOV3, a human OC cell line, was purchased from PerkinElmer (Waltham, MA, USA).

Techniques: Control

Journal: Pharmaceutics

Article Title: Synergistic Encapsulation of Paclitaxel and Sorafenib by Methoxy Poly(Ethylene Glycol)- b -Poly(Caprolactone) Polymeric Micelles for Ovarian Cancer Therapy

doi: 10.3390/pharmaceutics15041206

Figure Lengend Snippet: ( A ) Representative IVIS images of an SKOV3-red-fluc cell xenograft model taken 20 days after IV injection with DPBS (control), solution, and micelle, ( B ) relative total flux (% of day 9) value graph of each group (* p < 0.05, n = 5), ( C ) body weight change, ( D ) representative H&E staining of xenograft mouse model.

Article Snippet: SKOV3-red-fluc, a cancer cell line in which a fluorescent factor was introduced into SKOV3, a human OC cell line, was purchased from PerkinElmer (Waltham, MA, USA).

Techniques: IV Injection, Control, Staining

Journal: Molecular Cancer

Article Title: Systematic analyses reveal long non-coding RNA (PTAF)-mediated promotion of EMT and invasion-metastasis in serous ovarian cancer

doi: 10.1186/s12943-018-0844-7

Figure Lengend Snippet: miR-25 knockdown promotes OvCa cell migration. a The expression of miR-25 in OvCa cells after treatment with 10 ng/ml TGF-β1 for 48 h. b The mRNA levels of EMT-related markers in SKOV3 cells transfected with a miR-25 inhibitor (AMO-25) or a negative control (NC). c Western blotting analysis of epithelial and mesenchymal markers in SKOV3 cells transfected with AMO-25. Wound-healing ( d ) and migration assays ( e ) were used to determine the effect of miR-25 inhibition on OvCa cell migration. n = 5 independent experiments. * P < 0.05, ** p < 0.01 vs. NC

Article Snippet: In accordance with the manufacturer’s instructions, stable PTAF knockdown or overexpression SKOV3 cell lines and respective control cell lines were generated using lentiviral vectors carrying either sh-PTAF, a PTAF overexpression construct, or the respective negative controls (Biowit Technology, Shenzhen, China).

Techniques: Knockdown, Migration, Expressing, Transfection, Negative Control, Western Blot, Inhibition

Journal: Molecular Cancer

Article Title: Systematic analyses reveal long non-coding RNA (PTAF)-mediated promotion of EMT and invasion-metastasis in serous ovarian cancer

doi: 10.1186/s12943-018-0844-7

Figure Lengend Snippet: Forced expression of miR-25 blunts TGF-β1-induced EMT and migration in OvCa cells. qRT-PCR ( a ) and western blot ( b ) analyses showed the inhibitory effect of miR-25 on EMT in SKOV3 cells treated with TGF-β1. A wound-healing assay displayed the inhibitory effects of miR-25 on TGF-β1-induced migration in SKOV3 cells ( c ) and in A2780 cells ( d ). A migration assay showed that miR-25 attenuated TGF-β1-induced migration in SKOV3 cells ( e ) and A2780 cells ( f ). n = 5 independent experiments. * P < 0.05, ** P < 0.01

Article Snippet: In accordance with the manufacturer’s instructions, stable PTAF knockdown or overexpression SKOV3 cell lines and respective control cell lines were generated using lentiviral vectors carrying either sh-PTAF, a PTAF overexpression construct, or the respective negative controls (Biowit Technology, Shenzhen, China).

Techniques: Expressing, Migration, Quantitative RT-PCR, Western Blot, Wound Healing Assay

Journal: Molecular Cancer

Article Title: Systematic analyses reveal long non-coding RNA (PTAF)-mediated promotion of EMT and invasion-metastasis in serous ovarian cancer

doi: 10.1186/s12943-018-0844-7

Figure Lengend Snippet: PTAF regulates the expression and activity of miR-25. a PTAF contains a sequence domain complementary to the seed motif of miR-25. b Up-regulation of PTAF in OvCa cells treated with 10 ng/ml TGF-β1 for 48 h. n = 6 independent experiments. ** P < 0.01 vs. Control. c - d Overexpression of PTAF inhibited the expression of miR-25 in SKOV3 cells. n = 6 independent experiments. ** P < 0.01 vs. pcDNA3.1. e - f Silencing of PTAF using a specific shRNA up-regulated miR-25 in SKOV3 cells. n = 6 independent experiments. * P < 0.05, ** P < 0.01 vs. sh-Scramble. g - h PTAF binds to miR-25 and regulates its activity. * P < 0.05, ** P < 0.01. i PTAF directly binding to miR-25. SKOV3 cells were transfected with biotin-tagged miR-Ctrl (Bio-miR-Ctrl) or biotin-tagged miR-25 (Bio-miR-25). Forty-eight hours after transfection, the cells were harvested for a biotin-based pull-down assay. PTAF expression levels were analyzed by qRT-PCR. ** P < 0.01. j Luciferase reporter activities of chimeric vectors carrying the luciferase gene and a fragment of PTAF containing the wild-type (WT) binding site or a mutated binding site for miR-25. ** P < 0.01. k - l qRT-PCR were used to examine the expression of PTAF in SKOV3 cells after overexpression or knockdown of miR-25. * P < 0.05, ** P < 0.01. n = 6 independent experiments

Article Snippet: In accordance with the manufacturer’s instructions, stable PTAF knockdown or overexpression SKOV3 cell lines and respective control cell lines were generated using lentiviral vectors carrying either sh-PTAF, a PTAF overexpression construct, or the respective negative controls (Biowit Technology, Shenzhen, China).

Techniques: Expressing, Activity Assay, Sequencing, Control, Over Expression, shRNA, Binding Assay, Transfection, Pull Down Assay, Quantitative RT-PCR, Luciferase, Knockdown

Journal: Molecular Cancer

Article Title: Systematic analyses reveal long non-coding RNA (PTAF)-mediated promotion of EMT and invasion-metastasis in serous ovarian cancer

doi: 10.1186/s12943-018-0844-7

Figure Lengend Snippet: PTAF silencing inhibits tumor progression in an orthotopic mouse model of OvCa. a Representative bioluminescence images of mice injected intraperitoneally with SKOV3 OvCa cells expressing either PTAF or empty pcDNA3.1 ( n = 10 in each group). b More tumor masses (yellow arrows) were formed by SKOV3-PTAF cells than by SKOV3-pcDNA3.1 cells. c Mice injected with SKOV3-PTAF cells showed liver metastases. d Representative images of tumor nodules and quantification of tumor nodules and tumor weights in mice injected intraperitoneally with SKOV3 OvCa cells expressing either PTAF or empty pcDNA3.1 ( n = 10 in each group). ** P < 0.01 vs. pcDNA3.1. e SKOV3-ip tumor samples from mice injected with cells expressing PTAF or empty pcDNA3.1 were immunohistochemically stained for E-cadherin and SNAI2. * P < 0.05 vs. pcDNA3.1. f Representative bioluminescence images in sh-Scramble- and sh-PTAF-treated mice ( n = 10 in each group) that were injected intraperitoneally with SKOV3 OvCa cells. g Fewer tumor masses (yellow arrows) were formed in the mice treated with sh-PTAF than in those treated with sh-Scramble. h Mice injected with sh-Scramble showed liver metastases, whereas very little metastasis was observed in mice injected with sh-PTAF. i Representative images of tumor nodules and quantification of tumor nodules and tumor weights in sh-Scramble- and sh-PTAF-treated mice injected with SKOV3 cells ( n = 10 for each group). ** P < 0.01 vs. sh-Scramble. j SKOV3-ip tumor samples from sh-Scramble- and sh-PTAF-treated mice were immunohistochemically stained for E-cadherin and SNAI2. * P < 0.05 vs. sh-Scramble. Scale bars, 100X = 100 μm; 400X = 20 μm

Article Snippet: In accordance with the manufacturer’s instructions, stable PTAF knockdown or overexpression SKOV3 cell lines and respective control cell lines were generated using lentiviral vectors carrying either sh-PTAF, a PTAF overexpression construct, or the respective negative controls (Biowit Technology, Shenzhen, China).

Techniques: Injection, Expressing, Staining

Journal: Oxidative Medicine and Cellular Longevity

Article Title: HSP90AB1 as the Druggable Target of Maggot Extract Reverses Cisplatin Resistance in Ovarian Cancer

doi: 10.1155/2023/9335440

Figure Lengend Snippet: ME enhances the sensitivity of A2780/CDDP and SKOV3/CDDP cells to cisplatin (Cis). Cells were treated with different concentrations of cisplatin with and without ME (6 mg/ml) for 48 h. (a) The viability of A2780 and A2780/CDDP cells was assayed using CCK8 kits. (b) The viability of SKOV3 and SKOV3/CDDP cells was assayed. SKOV3/CDDP cells were infected with lentiviral particles to express luciferase, and then cells were treated with cisplatin (3.2 μ g/ml) and/or ME (4 mg/ml or 6 mg/ml) for 48 h followed by 150 μ g/ml D-luciferin for 10 min. (c) The luciferase-positive SKOV3/CDDP cells were analyzed using Living Image software and a GloMax® 96 Microplate Luminometer. The quantification of luciferase-positive cells is shown on the right. ∗ p ≤ 0.05 vs. blank; # p ≤ 0.05 vs. cisplatin; (d) A2780/CDDP and SKOV3/CDDP cells were double stained with Annexin V-FITC and PI then analyzed using flow cytometry, and the quantitative analysis was located in the right. ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001. (e) Levels of p-p53 in A2780/DDP cells were measured by immunofluorescence staining (400x magnification). The panel under the staining shows the quantitative analysis of p-p53 in A2780/CDDP cells. The nuclei are stained with DAPI. ∗ p ≤ 0.05 vs. blank; # p ≤ 0.05 vs. cisplatin. (f) Representative images of the wound healing assays in A2780/CDDP and SKOV3/CDDP cells are shown, and the panels under the images show the wound closure rate. ∗ p ≤ 0.05 vs. 0 h; # p ≤ 0.05 vs. blank; ns means no significance vs. 0 h. (g) The transwell assay was used to analyze the invasion of A2780 and A2780/CDDP cells. The invasive cell number was quantified using ImageJ and located under the images. ∗ p ≤ 0.05. Three independent experiments were performed with similar results. Data are shown as the mean ± SEM.

Article Snippet: The human ovarian cancer cell line A2780 (RRID: CVCL_0134) was purchased from Shanghai Pituo Biological Technology Co., Ltd. (China), and the cell line SKOV3 (RRID: CVCL_0532) was purchased from Solarbio Science & Technology Co., Ltd. (China).

Techniques: Infection, Luciferase, Software, Staining, Flow Cytometry, Immunofluorescence, Transwell Assay

Journal: Oxidative Medicine and Cellular Longevity

Article Title: HSP90AB1 as the Druggable Target of Maggot Extract Reverses Cisplatin Resistance in Ovarian Cancer

doi: 10.1155/2023/9335440

Figure Lengend Snippet: Enrichment analysis of DEGs in A2780/CDDP cells based on high-throughput RNA sequencing. The total RNA from A2780 and A2780/DDP cells was extracted using RNA Miniprep kit reagents. Next-generation sequencing analysis was performed on the BGISEQ-500 platform by BGI Genomic Services. (a) Volcano plot showing significantly upregulated genes (red dots) and downregulated genes (blue dots). (b) A2780 vs. A2780/CDDP Gene Ontology (GO) analysis on a cellular component of DEGs. (c) A2780 vs. A2780/CDDP KEGG pathway enrichment analysis of DEGs. (d) The heat map shows the relative transcript levels of the DEGs in A2780 and A2780/CDDP cells. (e) The protein–protein interaction network shows the upregulated DEGs from A2780/CDDP cells compared with A2780 cells. (f) qRT-PCR analyses of the mRNA levels of MYC in A2780 cells and A2780/CDDP cells. (g) qRT-PCR analyses of the mRNA levels of HSP90AB1 in A2780 cells and A2780/CDDP cells. Three independent experiments were performed with similar results. Data are shown as the mean ± SEM. ∗ p ≤ 0.05, ∗∗∗ p ≤ 0.001.

Article Snippet: The human ovarian cancer cell line A2780 (RRID: CVCL_0134) was purchased from Shanghai Pituo Biological Technology Co., Ltd. (China), and the cell line SKOV3 (RRID: CVCL_0532) was purchased from Solarbio Science & Technology Co., Ltd. (China).

Techniques: High Throughput Screening Assay, RNA Sequencing, Next-Generation Sequencing, Quantitative RT-PCR

Journal: Oxidative Medicine and Cellular Longevity

Article Title: HSP90AB1 as the Druggable Target of Maggot Extract Reverses Cisplatin Resistance in Ovarian Cancer

doi: 10.1155/2023/9335440

Figure Lengend Snippet: ME induces cisplatin to promote apoptosis by suppressing the expression of HSP90AB1/IGF1R in cisplatin-resistant ovarian cancer cells. (a) The expression of HSP90AB1, IGF1R, and IGFBP2 in A2780 cells and A2780/CDDP cells was assessed by western blot. (b) The lower panel shows the quantitative analysis. ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001. (c) The expression of HSP90AB1, IGF1R, and IGFBP2 in SKOV3 cells and SKOV3/CDDP cells was assessed by western blot. (d) The lower panel shows the quantitative analysis. ∗ p ≤ 0.05, ∗∗ p ≤ 0.01. The GSH level in A2780 cells (e) and SKOV3 cells (f) was analyzed using a reduced GSH assay kit. ∗ p ≤ 0.05, ns means no significance. Relative mRNA expression of MYC (g), HSP90AB1 (h), and IGF1R (i) in A2780/CDDP cells was determined by real-time PCR. ∗ p ≤ 0.05, ∗∗∗ p ≤ 0.001, ns means no significance. A2780/CDDP and SKOV3/CDDP cells were treated with cisplatin (3.2 μ g/ml) and ME (6 mg/ml) for 48 h. The protein was extracted for further analysis. (j–m) The protein expression of HSP90AB1, IGF1R, MYC, PTEN, p-H2AX, and BCL2 in A2780/CDDP cells (j) and SKOV3/CDDP cells (l) was measured by western blot. The lower panel shows the quantitative analysis of the western blot in A2780/CDDP cells (K) and SKOV3/CDDP cells (m). ∗ p ≤ 0.05, ns means no significance. Drug-resistant cells were treated with cisplatin, ME, or both. The amount of GSH in A2780/CDDP cells (n) and SKOV3/CDDP cells (o) was analyzed using a reduced GSH assay kit. ∗ p ≤ 0.05, ns means no significance. Three independent experiments were performed with similar results. Data are shown as the mean ± SEM.

Article Snippet: The human ovarian cancer cell line A2780 (RRID: CVCL_0134) was purchased from Shanghai Pituo Biological Technology Co., Ltd. (China), and the cell line SKOV3 (RRID: CVCL_0532) was purchased from Solarbio Science & Technology Co., Ltd. (China).

Techniques: Expressing, Western Blot, GSH Assay, Real-time Polymerase Chain Reaction

Journal: Oxidative Medicine and Cellular Longevity

Article Title: HSP90AB1 as the Druggable Target of Maggot Extract Reverses Cisplatin Resistance in Ovarian Cancer

doi: 10.1155/2023/9335440

Figure Lengend Snippet: Inhibition of HSP90 ATPase activity with geldanamycin promotes apoptosis and suppresses migration in cisplatin-resistant ovarian cancer cells. A2780/CDDP cells and SKOV3/CDDP cells were pretreated with 5 μ M geldanamycin (In-HSP) for 1 h and then treated with cisplatin (3.2 μ g/ml) and ME (6 mg/ml) for 72 h. (a) A2780/CDDP cells and SKOV3/CDDP cells were double stained with Annexin V-FITC and PI and then analyzed by flow cytometry, and the quantitative analysis was located on the right. ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001. (b, c) A2780/CDDP cells and SKOV3/CDDP cells expressing luciferase were pretreated with 150 μ g/ml D-luciferin for 10 min. The luciferase-positive A2780/CDDP cells (b) and SKOV3/CDDP cells (c) were analyzed using Living Image software and a GloMax® 96 Microplate Luminometer. The quantification of luciferase-positive cells is shown on the right. ∗∗∗ p ≤ 0.001, ∗∗∗∗ p ≤ 0.0001. Representative images of the wound healing assays of A2780/CDDP cells (d) and SKOV3/CDDP cells (e) are shown, and the quantification of the wound closure rate is on the right. ∗ p ≤ 0.05 vs. 0 h; ns mean no significance vs. 0 h; ## p ≤ 0.01. (f) The expression of HSP90AB1, IGF1R, p-H2AX, p-p53, and BAX in A2780/CDDP cells was analyzed by western blot. (g) The panel on the right shows the quantitative analysis. ∗ p ≤ 0.05. (h) The expression of HSP90AB1, IGF1R, p-H2AX, p-p53, and BCL2 in SKOV3/CDDP cells was measured by western blot. (i) The panel on the right shows the quantitative analysis. ∗ p ≤ 0.05. (j–m) The coimmunoprecipitation assay. SKOV3/CDDP cells were treated with geldanamycin for 24 h. The expression levels of HSP90AB1 and IGF1R were measured (j), and the panel on the right shows the quantitative analysis (k). ∗ p ≤ 0.05. The same cell lysates were immunoprecipitated with IGF1R, HSP90AB1, and isoform-matched immunoglobulin (IgG). (l) Western blot assays of SKOV3/CDDP cells using site-specific antibodies against HSP90AB1 and IGF1R, and the panel on the right shows the quantitative analysis (m). ∗ p ≤ 0.05. Three independent experiments were performed with similar results. Data are shown as the mean ± SEM.

Article Snippet: The human ovarian cancer cell line A2780 (RRID: CVCL_0134) was purchased from Shanghai Pituo Biological Technology Co., Ltd. (China), and the cell line SKOV3 (RRID: CVCL_0532) was purchased from Solarbio Science & Technology Co., Ltd. (China).

Techniques: Inhibition, Activity Assay, Migration, Staining, Flow Cytometry, Expressing, Luciferase, Software, Western Blot, Co-Immunoprecipitation Assay, Immunoprecipitation