Journal: Future science OA

Article Title: Adverse predictive value of ASPM on lung adenocarcinoma overall survival depended on chemotherapy status.

doi: 10.1080/20565623.2025.2489328

Figure Lengend Snippet: Figure 1. (A) TCGA TARGET GTEx pan-cancer dataset analysis showed the relationship between ASPM gene expression and patient prognosis in each tumor. (B) In TCGA dataset of LUAD, high ASPM expression was related to worse overall survival (Log-rank test p = 2.4e-6). (C) In proteomics dataset of LUAD, the difference of prognosis was not significant between ASPM high or low expression groups (Log-rank test p = 0.26).

Article Snippet: Immunohistochemistry and special staining the Luad tissues (n = 160) were obtained from surgically resected specimens and made into tissue microarray. the sections were de-paraffinized and hydrated, and the endogenous peroxidase was blocked. antigen retrieval was performed using the dako target Retrieval Solution, High pH (dako ominis, agilent technologies, Santa Clara, Ca, uSa), in a ptLink set at 98 °C for 25 min. the tissue sections were incubated with an anti-aSpm rabbit polyclonal antibody (1:200, a02584-1, Boster, China) 1h at 37 °C. detection of immunostaining was achieved by an enzyme-conjugated polymer complex (K8002, dako, denmark) adapted for autostainers from daKo (dako autostainer, agilent technologies). the staining intensity in selected areas was scored as follows: 0, no staining; 1, weak staining; 2, moderate staining; 3, strong staining. the stained area was scored as follows: 0, no staining; 1, 1–10% positive cells; 2, 10–50% positive cells; 3, > 50% positive cells. the final score was evaluated by multiplying the staining intensity and stained area percentage. the negative and weak staining cases were considered as low expression group while the moderate and strong staining cases were defined as high expression group in later analysis.

Techniques: Gene Expression, Expressing

Journal: Future science OA

Article Title: Adverse predictive value of ASPM on lung adenocarcinoma overall survival depended on chemotherapy status.

doi: 10.1080/20565623.2025.2489328

Figure Lengend Snippet: Figure 2. (A) The representative immunohistochemical staining of ASPM negative, weak, moderate, and strong cases. (B) In our cohort, high ASPM expression predicted better LUAD prognosis (Log-rank test p = 5.4e-4). (C) In patients who received chemotherapy, high ASPM expression was related to better overall survival (Log-rank test p = 5.0e-4). (D) In patients who did not received chemotherapy, ASPM expression was not related to patient overall survival (Log-rank test p = 0.48). (E) Multivariate regression analysis showed ASPM was not an independent prognostic factor while other protective factor included chemother apy and risk factor included clinical stages. (F) time dependent ROC analysis was conducted at time points 365, 1095, and 1825 days. The AUC and confidence intervals were evaluated using the ci function of pROC.

Article Snippet: Immunohistochemistry and special staining the Luad tissues (n = 160) were obtained from surgically resected specimens and made into tissue microarray. the sections were de-paraffinized and hydrated, and the endogenous peroxidase was blocked. antigen retrieval was performed using the dako target Retrieval Solution, High pH (dako ominis, agilent technologies, Santa Clara, Ca, uSa), in a ptLink set at 98 °C for 25 min. the tissue sections were incubated with an anti-aSpm rabbit polyclonal antibody (1:200, a02584-1, Boster, China) 1h at 37 °C. detection of immunostaining was achieved by an enzyme-conjugated polymer complex (K8002, dako, denmark) adapted for autostainers from daKo (dako autostainer, agilent technologies). the staining intensity in selected areas was scored as follows: 0, no staining; 1, weak staining; 2, moderate staining; 3, strong staining. the stained area was scored as follows: 0, no staining; 1, 1–10% positive cells; 2, 10–50% positive cells; 3, > 50% positive cells. the final score was evaluated by multiplying the staining intensity and stained area percentage. the negative and weak staining cases were considered as low expression group while the moderate and strong staining cases were defined as high expression group in later analysis.

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Future science OA

Article Title: Adverse predictive value of ASPM on lung adenocarcinoma overall survival depended on chemotherapy status.

doi: 10.1080/20565623.2025.2489328

Figure Lengend Snippet: Figure 3. (A) heatmap of signature proteins in ASPM low or high groups from proteomics dataset. (B) High ASPM1 related signature proteins were enriched mainly in cell cycle and mitosis processes by Metascape analysis. (C) ASPM positive group in proteomics dataset showed higher expression of proliferation related proteins mainly including AURKB, PLK1, KIFC3, EZH2,CDKN2A, BRD3, CASP2, MKI67, CDK1, and MDC1.

Article Snippet: Immunohistochemistry and special staining the Luad tissues (n = 160) were obtained from surgically resected specimens and made into tissue microarray. the sections were de-paraffinized and hydrated, and the endogenous peroxidase was blocked. antigen retrieval was performed using the dako target Retrieval Solution, High pH (dako ominis, agilent technologies, Santa Clara, Ca, uSa), in a ptLink set at 98 °C for 25 min. the tissue sections were incubated with an anti-aSpm rabbit polyclonal antibody (1:200, a02584-1, Boster, China) 1h at 37 °C. detection of immunostaining was achieved by an enzyme-conjugated polymer complex (K8002, dako, denmark) adapted for autostainers from daKo (dako autostainer, agilent technologies). the staining intensity in selected areas was scored as follows: 0, no staining; 1, weak staining; 2, moderate staining; 3, strong staining. the stained area was scored as follows: 0, no staining; 1, 1–10% positive cells; 2, 10–50% positive cells; 3, > 50% positive cells. the final score was evaluated by multiplying the staining intensity and stained area percentage. the negative and weak staining cases were considered as low expression group while the moderate and strong staining cases were defined as high expression group in later analysis.

Techniques: Expressing

Journal: Future science OA

Article Title: Adverse predictive value of ASPM on lung adenocarcinoma overall survival depended on chemotherapy status.

doi: 10.1080/20565623.2025.2489328

Figure Lengend Snippet: Figure 4. (A) Proteomics data analysis showed ASPM positive group had higher MKI67 expression. (B) Western blot analysis of A549 cells and H1299 cells confirmed effectiveness of ASPM transfection and in vitro overexpression. (C-D) Statistical anal ysis of western blot images. (E-F) CCK8 assay proved overexpression of ASPM promoted tumor cell proliferation in A549 and H1299 cells. (G-H) When treated with cisplatin, overexpression of ASPM enhanced the lethality on A549 and H1299 tumor cells. (P value *<0.05, **<0.01, ***<0.001).

Article Snippet: Immunohistochemistry and special staining the Luad tissues (n = 160) were obtained from surgically resected specimens and made into tissue microarray. the sections were de-paraffinized and hydrated, and the endogenous peroxidase was blocked. antigen retrieval was performed using the dako target Retrieval Solution, High pH (dako ominis, agilent technologies, Santa Clara, Ca, uSa), in a ptLink set at 98 °C for 25 min. the tissue sections were incubated with an anti-aSpm rabbit polyclonal antibody (1:200, a02584-1, Boster, China) 1h at 37 °C. detection of immunostaining was achieved by an enzyme-conjugated polymer complex (K8002, dako, denmark) adapted for autostainers from daKo (dako autostainer, agilent technologies). the staining intensity in selected areas was scored as follows: 0, no staining; 1, weak staining; 2, moderate staining; 3, strong staining. the stained area was scored as follows: 0, no staining; 1, 1–10% positive cells; 2, 10–50% positive cells; 3, > 50% positive cells. the final score was evaluated by multiplying the staining intensity and stained area percentage. the negative and weak staining cases were considered as low expression group while the moderate and strong staining cases were defined as high expression group in later analysis.

Techniques: Expressing, Western Blot, Transfection, In Vitro, Over Expression, CCK-8 Assay

Journal: Frontiers in Pharmacology

Article Title: Antitumor Activity of Abnormal Cannabidiol and Its Analog O-1602 in Taxol-Resistant Preclinical Models of Breast Cancer

doi: 10.3389/fphar.2019.01124

Figure Lengend Snippet: Cell viability. (A) Effect of paclitaxel on sensitive and resistant (PR) MCF-7 and MDA-MB-231 cells. (B) Effect of O-1602 and Abn-CBD on the cell viability of MCF-10A cells. (C) Cell viability measured in paclitaxel-sensitive MCF-7 cells following 48-h treatment with various concentrations of O-1602 alone or in presence of paclitaxel. (D) Effect of Abn-CBD ± paclitaxel on taxol-sensitive MCF-7 cells. (E) Effect of O-1602 ± paclitaxel on taxol-sensitive MDA-MB-231 cells. (F) Effect of Abn-CBD ± paclitaxel on taxol-sensitive MDA-MB-231 cells.

Article Snippet: Cells were grown on glass coverslips in six-well plates and then treated with DMSO or 2 μM O-1602 or abnormal CBD in DMSO for 24 h. The annexin V apoptosis detection kit (Santa Cruz Biotechnology) was used to determine the rate of apoptosis.

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Antitumor Activity of Abnormal Cannabidiol and Its Analog O-1602 in Taxol-Resistant Preclinical Models of Breast Cancer

doi: 10.3389/fphar.2019.01124

Figure Lengend Snippet: Atypical cannabinoids effect on the viability of paclitaxel-resistant cells. (A) Cell viability measured in paclitaxel-resistant (PR) MCF-7 cells following 48-h treatment with various concentrations of O-1602 alone or in presence of paclitaxel. (B) Effect of Abn-CBD ± paclitaxel on MCF-7 PR cells. (C) Effect of O-1602 ± paclitaxel on MDA-MB-231 PR cells. (D) Effect of Abn-CBD ± paclitaxel on MDA-MB-231 PR cells.

Article Snippet: Cells were grown on glass coverslips in six-well plates and then treated with DMSO or 2 μM O-1602 or abnormal CBD in DMSO for 24 h. The annexin V apoptosis detection kit (Santa Cruz Biotechnology) was used to determine the rate of apoptosis.

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Antitumor Activity of Abnormal Cannabidiol and Its Analog O-1602 in Taxol-Resistant Preclinical Models of Breast Cancer

doi: 10.3389/fphar.2019.01124

Figure Lengend Snippet: Effect of atypical cannabinoids on apoptosis. Cells were treated for 24 h with either the DMSO vehicle, O-1602, or Abn-CBD. (A) Histogram showing the % of annexin V–labeled cells. Cells staining for PI only or both PI/AV are not shown. Cells were counted from three random fields of view on a fluorescence microscope. * p < 0.05, ** p < 0.01, n = 3. (B) Western blotting analysis was performed using an anti–caspase-3 antibody, and β-tubulin was included as a loading control. Figure is a representative blot of n = 3 experiments. (C) Reactive oxygen species assay measurements in MDA-MB-231 and MCF-7 cells following treatment with O-1602 or Abn-CBD. (n = 3).

Article Snippet: Cells were grown on glass coverslips in six-well plates and then treated with DMSO or 2 μM O-1602 or abnormal CBD in DMSO for 24 h. The annexin V apoptosis detection kit (Santa Cruz Biotechnology) was used to determine the rate of apoptosis.

Techniques: Labeling, Staining, Fluorescence, Microscopy, Western Blot, Control

Journal: Frontiers in Pharmacology

Article Title: Antitumor Activity of Abnormal Cannabidiol and Its Analog O-1602 in Taxol-Resistant Preclinical Models of Breast Cancer

doi: 10.3389/fphar.2019.01124

Figure Lengend Snippet: Effect of atypical cannabinoids on EGF-mediated migration and invasion. (A) Histogram summarizing Transwell migration results using MDA-MB-231 PR cells in response to EGF in presence of either the vehicle control or various concentrations of O-1602 or Abn-CBD. (B) Histogram summarizing Matrigel invasion using MDA-MB-231 PR cells in presence of either the vehicle control, O-1602, or Abn-CBD. Results represent the means ± SEM of at least three independent experiments. * p < 0.05, ** p < 0.01.

Article Snippet: Cells were grown on glass coverslips in six-well plates and then treated with DMSO or 2 μM O-1602 or abnormal CBD in DMSO for 24 h. The annexin V apoptosis detection kit (Santa Cruz Biotechnology) was used to determine the rate of apoptosis.

Techniques: Migration, Control

Journal: Frontiers in Pharmacology

Article Title: Antitumor Activity of Abnormal Cannabidiol and Its Analog O-1602 in Taxol-Resistant Preclinical Models of Breast Cancer

doi: 10.3389/fphar.2019.01124

Figure Lengend Snippet: Toxicity of atypical cannabinoids in zebrafish. Graph showing the percentage of zebrafish larvae affected by various concentrations of O-1602 or Abn-CBD in water for 3 days.

Article Snippet: Cells were grown on glass coverslips in six-well plates and then treated with DMSO or 2 μM O-1602 or abnormal CBD in DMSO for 24 h. The annexin V apoptosis detection kit (Santa Cruz Biotechnology) was used to determine the rate of apoptosis.

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Antitumor Activity of Abnormal Cannabidiol and Its Analog O-1602 in Taxol-Resistant Preclinical Models of Breast Cancer

doi: 10.3389/fphar.2019.01124

Figure Lengend Snippet: Effect of atypical cannabinoids on the viability of Paclitaxel-resistant breast cancer cells in vivo . (A) Images of representative zebrafish injected with human MDA-MB-231 PR cells in the yolk sac (white pixels) before (0 hpf) and following 3-day treatment with the DMSO vehicle control, O-1602, or Abn-CBD (72 hpf). (B) Quantification of the analysis of more than 24 images for each condition. *** p < 0.001.

Article Snippet: Cells were grown on glass coverslips in six-well plates and then treated with DMSO or 2 μM O-1602 or abnormal CBD in DMSO for 24 h. The annexin V apoptosis detection kit (Santa Cruz Biotechnology) was used to determine the rate of apoptosis.

Techniques: In Vivo, Injection, Control