Journal: Current Issues in Molecular Biology

Article Title: Associating Patient Responses with Drug Sensitivity in Non-Small Cell Lung Carcinoma Using an Immunoassay on Patient-Derived Cell Cultures

doi: 10.3390/cimb47040281

Figure Lengend Snippet: Clinical parameters of NSCLC patients included in our study.

Article Snippet: Vinorelbine, pemetrexed, and osimertinib were purchased from Selleckchem (Houston, TX, USA).

Techniques: Mutagenesis

Journal: International journal of cancer

Article Title: SOCS-1 gene delivery cooperates with cisplatin plus pemetrexed to exhibit preclinical antitumor activity against malignant pleural mesothelioma.

doi: 10.1002/ijc.27611

Figure Lengend Snippet: Figure 2. Effect of AdSOCS-1, cisplatin and pemetrexed on viability of MPM cell lines (MESO-4, H28 and H226). (a) Effect of AdSOCS-1 on viability of three MPM cell lines. Cells were infected with either AdSOCS-1 or AdLacZ. After a 72-hr culture, viable cell numbers were counted by MTS assay. Values shown represent means þ SD of triplicate wells. (b) Effect of cisplatin and pemetrexed on viability of three MPM cell lines. Cells were cultivated in the presence of cisplatin (1–100 lM) or pemetrexed (10–5,000 lM). After 72 hr culture, cell viability was estimated using the MTS assay. Values shown represent means 6 SD of triplicate wells. (c) Effect of combined AdSOCS-1 and cisplatin plus pemetrexed on viability of three MPM cell lines. Cells were infected with either AdSOCS-1 or AdLacZ as control at an MOI of 40. After 6 hr from infection, cells were cultivated with or without 10 lM cisplatin (C) plus 200 lM pemetrexed (P). After an additional 120- hr culture, cell viability was estimated using the MTS assay. Values shown represent means þ SD of triplicate wells.

Article Snippet: Transfected cells were treated with cisplatin and pemetrexed in serumfree medium after 6 hr, and invasion assay was performed 24 hr after treatment using a Cultrex 96-well membrane invasion assay kit (R&D Systems, Minneapolis, MN) according to the manufacturer’s instructions.

Techniques: Infection, MTS Assay, Control

Journal: International journal of cancer

Article Title: SOCS-1 gene delivery cooperates with cisplatin plus pemetrexed to exhibit preclinical antitumor activity against malignant pleural mesothelioma.

doi: 10.1002/ijc.27611

Figure Lengend Snippet: Figure 3. Apoptosis and cell invasion regulated by AdSOCS-1 combined with cisplatin and pemetrexed in MPM cell lines (MESO-4, H28 and H226). (a) Apoptosis assay. MPM cells were infected with either AdSOCS-1 or AdLacZ as control at an MOI of 40. After 6 hr from infection, cells were cultivated with or without 10 lM cisplatin (C) plus 200 lM pemetrexed (P). After an additional 72-hr culture, apoptosis was determined by means of annexin V and 7-AAD staining using flow cytometry. Values shown represent means þ SD of triplicate wells. NS means not significant. (b) Cell invasion assay. MPM cells were infected with either AdSOCS-1 or AdLacZ as control at an MOI of 40. After 6 hr from infection, transfected cells were treated with or without 10 lM cisplatin (C) and 200 lM pemetrexed (P) in serum-free medium after 6 hr and invasion assay was performed 24 hr after treatment. Values shown represent means þ SD of triplicate wells. NS means not significant.

Article Snippet: Transfected cells were treated with cisplatin and pemetrexed in serumfree medium after 6 hr, and invasion assay was performed 24 hr after treatment using a Cultrex 96-well membrane invasion assay kit (R&D Systems, Minneapolis, MN) according to the manufacturer’s instructions.

Techniques: Apoptosis Assay, Infection, Control, Staining, Cytometry, Invasion Assay, Transfection

Journal: International journal of cancer

Article Title: SOCS-1 gene delivery cooperates with cisplatin plus pemetrexed to exhibit preclinical antitumor activity against malignant pleural mesothelioma.

doi: 10.1002/ijc.27611

Figure Lengend Snippet: Figure 4. Caspase pathway activated by combined AdSOCS-1 and cisplatin plus pemetrexed. (a) Inhibition of antiapoptotic proteins. Three MPM cell lines (MESO-4, H28 and H226) were infected with either AdSOCS-1 or AdLacZ as control at an MOI of 40. After 6 hr from infection, cells were cultivated with or without 10 lM cisplatin (C) plus 200 lM pemetrexed (P). After an additional 24-hr (upper panel) or 48-hr (lower panel) culture, protein extracts were blotted with indicated antibodies. (b) Activation of caspases. MESO-4 and H226 cells were infected with either AdSOCS-1 or AdLacZ as control at an MOI of 40. After 6 hr from infection, cells were cultivated with or without 10 lM cisplatin (C) plus 200 lM pemetrexed (P). After an additional 72-hr culture, protein extracts were blotted with indicated antibodies. (c) Cleavage of FAK and Akt by caspase-3. Protein extracts of MESO-4 cells were incubated with recombinant active caspase-3 with or without z-DEVD-fmk. After 3 hr of incubation, protein extracts were blotted with indicated antibodies.

Article Snippet: Transfected cells were treated with cisplatin and pemetrexed in serumfree medium after 6 hr, and invasion assay was performed 24 hr after treatment using a Cultrex 96-well membrane invasion assay kit (R&D Systems, Minneapolis, MN) according to the manufacturer’s instructions.

Techniques: Inhibition, Infection, Control, Activation Assay, Incubation, Recombinant

Journal: International journal of cancer

Article Title: SOCS-1 gene delivery cooperates with cisplatin plus pemetrexed to exhibit preclinical antitumor activity against malignant pleural mesothelioma.

doi: 10.1002/ijc.27611

Figure Lengend Snippet: Figure 5. NF-jB and STAT3 signaling is regulated by AdSOCS-1. (a, upper panel) Activation of NF-jB p65 by TNF-a. After 12 hr of serum starvation, MESO-4 cells were incubated with 10 ng/ml TNF-a for 0–90 min. Cytoplasmic and nuclear fraction of protein extracts were blotted with indicated antibodies. (a, lower panel) Inhibition of NF-jB by SOCS-1. MESO-4 and H226 cells were infected with either AdSOCS-1 or AdLacZ as control at an MOI of 40. After 6 hr from infection, cells were cultivated with or without 10 lM cisplatin (C) plus 200 lM pemetrexed (P). After an additional 24-hr culture in serum-starved medium, MESO-4 and H226 cells were stimulated with 10 ng/ml TNF-a for 30 min. Cytoplasmic and nuclear fraction of protein extracts were blotted with indicated antibodies. (b) Inhibition of STAT3 by SOCS-1. MESO-4 and H226 cells were infected with either AdSOCS-1 or AdLacZ as control at an MOI of 40. After 6 hr from infection, cells were cultivated with or without 10 lM cisplatin (C) plus 200 lM pemetrexed (P). After an additional 24-hr culture, protein extracts were blotted with indicated antibodies. (c) Inhibition of antiapoptotic proteins by NF-jB p65 and STAT3. STAT3, NF-jB p65 or nonspecific siRNA as control was added to MESO-4 and H226 cells. After 24-hr culture, protein extracts were blotted with indicated antibodies. (d) DNA binding and luciferase assay of NF-jB p65 (upper panel) and STAT3 (lower panel). MESO-4 and H226 cells were infected with either AdSOCS-1 or AdLacZ as control at an MOI of 40. After 6 hr from infection, cells were cultivated with or without 10 lM cisplatin (C) plus 200 lM pemetrexed (P). After an additional 24-hr culture, cells were analyzed by DNA binding and luciferase assay. Values shown represent means þ SD of triplicate wells. NS means not significant.

Article Snippet: Transfected cells were treated with cisplatin and pemetrexed in serumfree medium after 6 hr, and invasion assay was performed 24 hr after treatment using a Cultrex 96-well membrane invasion assay kit (R&D Systems, Minneapolis, MN) according to the manufacturer’s instructions.

Techniques: Activation Assay, Incubation, Inhibition, Infection, Control, Binding Assay, Luciferase

Journal: International journal of cancer

Article Title: SOCS-1 gene delivery cooperates with cisplatin plus pemetrexed to exhibit preclinical antitumor activity against malignant pleural mesothelioma.

doi: 10.1002/ijc.27611

Figure Lengend Snippet: Figure 6. Antitumor effect of AdSOCS-1 combined with cisplatin and pemetrexed in vivo. (a) Gross appearance of MESO-4 and H226 tumors grown orthotopically in the thoracic spaces. Female ICR nu/nu mice were intrathoracically treated with AdSOCS-1 or AdLacZ and intraperitoneally treated with pemetrexed and cisplatin after the implantation of MESO-4 or H226 cells into the pleural space. (b) Each tumor nodule found in the thoracic spaces was also weighed. Values shown represent means þ SD of five (MESO-4) or six (H226) mice. NS means not significant. (c) Immunohistochemical analysis of SOCS-1, NFjB p65 and TUNEL (blue fluorescence ¼ DAPI staining for nuclei; cyan fluorescence ¼ TUNEL positivity) in H226 tissue. The mice were treated in the same way as described above.

Article Snippet: Transfected cells were treated with cisplatin and pemetrexed in serumfree medium after 6 hr, and invasion assay was performed 24 hr after treatment using a Cultrex 96-well membrane invasion assay kit (R&D Systems, Minneapolis, MN) according to the manufacturer’s instructions.

Techniques: In Vivo, Immunohistochemical staining, TUNEL Assay, Staining

Journal: Lung cancer (Amsterdam, Netherlands)

Article Title: Cytotoxic chemotherapeutic agents and the EGFR-TKI osimertinib induce calreticulin exposure in non-small cell lung cancer.

doi: 10.1016/j.lungcan.2021.03.018

Figure Lengend Snippet: Fig. 4. Concentrations of soluble CRT in plasma of NSCLC patients before and after the onset of treatment with cytotoxic agents or osimertinib. (A) The plasma concentration of soluble CRT was determined before (pre), on days 3 and 8 of, and after the end of the first cycle of chemo therapy with DTX (n = 12) or PEM (n = 4) in patients with advanced NSCLC. The maximum (max) value for soluble CRT observed after the onset of treatment was then compared with the corresponding baseline value. **P < 0.01 (Wilcoxon matched-pairs signed-rank test). (B) The plasma concentration of soluble CRT was determined before (pre), on days 3 and 8 of, and about 1 month after the onset of osimertinib therapy in patients with EGFR-mutated advanced NSCLC (n = 9). The maximum (max) value for soluble CRT observed after the onset of treatment was then compared with the cor responding baseline value. ***P < 0.001 (Wil coxon matched-pairs signed-rank test).

Article Snippet: Pemetrexed disodium (PEM) was obtained from LKT Laboratories; docetaxel (DTX), cisplatin (CDDP), and carboplatin (CBDCA) were from Tocris; gemcitabine (GEM) was from Tronto; and paclitaxel (PTX), vinorelbine ditartrate (VNR), dimethyl sulfoxide were from Wako, and Z-VAD-FMK was from Selleck.

Techniques: Clinical Proteomics, Concentration Assay