Journal: Cells

Article Title: Molecular and Kinetic Analyses of Circulating Tumor Cells as Predictive Markers of Treatment Response in Locally Advanced Rectal Cancer Patients

doi: 10.3390/cells8070641

Figure Lengend Snippet: Immunostaining ( A–C ) and chromogenic in situ hybridization (CISH) ( D–F ) of CTCs from locally advanced rectal cancer (LARC) patients. ( A ) CTCs and leukocytes visualized with haematoxylin-eosin staining (×40 magnification). ( B ) CTCs stained with an anti-thymidylate synthase (TYMS) antibody, visualized with Permanent Red, and counterstained with haematoxylin (×40 magnification). ( C ) CTCs stained with an anti-RAD23B antibody, visualized with 3,3-diaminobenzidine (DAB), and counterstained with haematoxylin (×60 magnification). ( D ) CTCs negative for TYMS mRNA and counterstained with haematoxylin (×40 magnification). ( E ) CTCs with a low TYMS mRNA signal (normal expression) counterstained with haematoxylin (×40 magnification). ( F ) CTCs with a high TYMS mRNA signal (overexpression) counterstained with haematoxylin (×40 magnification). All images were analyzed on a Research System Microscope BX61 (Olympus, Tokyo, Japan) coupled to a digital camera (SC100–Olympus). Thick arrows indicate CTCs, thin arrows indicate leukocytes, and asterisks indicate 8 μm pores of the ISET ® membranes.

Article Snippet: Briefly, ISET membrane spots were cut out and subjected to immunocytochemistry (ICC) with an anti-RAD23B antibody (1:100 CSB-PA019260LA01HU; CusaBio, Wuhan, People’s Republic of China), an anti-TYMS antibody (1:230 WH0007298M1; Sigma-Aldrich, St. Louis, MO, USA), and an anti-CD45 antibody (1:200 HPA000440; Sigma-Aldrich).

Techniques: Immunostaining, Chromogenic In Situ Hybridization, Staining, Expressing, Over Expression, Microscopy

Journal: Cells

Article Title: Molecular and Kinetic Analyses of Circulating Tumor Cells as Predictive Markers of Treatment Response in Locally Advanced Rectal Cancer Patients

doi: 10.3390/cells8070641

Figure Lengend Snippet: Expression profiles of RAD23B and TYMS proteins and TYMS mRNA.

Article Snippet: Briefly, ISET membrane spots were cut out and subjected to immunocytochemistry (ICC) with an anti-RAD23B antibody (1:100 CSB-PA019260LA01HU; CusaBio, Wuhan, People’s Republic of China), an anti-TYMS antibody (1:230 WH0007298M1; Sigma-Aldrich, St. Louis, MO, USA), and an anti-CD45 antibody (1:200 HPA000440; Sigma-Aldrich).

Techniques: Expressing

Journal: Cells

Article Title: Molecular and Kinetic Analyses of Circulating Tumor Cells as Predictive Markers of Treatment Response in Locally Advanced Rectal Cancer Patients

doi: 10.3390/cells8070641

Figure Lengend Snippet: Summary of methodologies, analyses, and results in the present study. Patients were enrolled prior to the start of NCRT. Blood was collected to perform CTC counts and molecular analyses. At baseline, complete response (CR) correlated with low levels of TYMS mRNA. In contrast, NR correlated with high levels of TYMS mRNA and RAD23B protein expression. Blood samples were collected again during follow-up after NCRT. CTC analyses showed that CR correlated to low levels of TYMS mRNA and RAD23B protein expression, while NR correlated to high levels of TYMS mRNA and RAD23B/TYMS protein expression. CTC kinetics also correlated to pathological response. Based on these results, TYMS mRNA and RAD23B and TYMS protein expression appear to have a clinical and therapeutic impact in LARC patients. Abbreviations: CR: Complete response; LARC: Locally advanced rectal cancer; NCRT: Neoadjuvant chemoradiotherapy; NR: No response; PR: Partial response.

Article Snippet: Briefly, ISET membrane spots were cut out and subjected to immunocytochemistry (ICC) with an anti-RAD23B antibody (1:100 CSB-PA019260LA01HU; CusaBio, Wuhan, People’s Republic of China), an anti-TYMS antibody (1:230 WH0007298M1; Sigma-Aldrich, St. Louis, MO, USA), and an anti-CD45 antibody (1:200 HPA000440; Sigma-Aldrich).

Techniques: Expressing

Journal: Thoracic Cancer

Article Title: Knockdown of circ‐RAD23B inhibits non‐small cell lung cancer progression via the miR ‐142‐3p/ MAP4K3 axis

doi: 10.1111/1759-7714.14319

Figure Lengend Snippet: Correlation between clinicopathological parameters of NSCLC patients and circ‐RAD23B expression

Article Snippet: A549 cells infected with lentivirus sh‐circ‐RAD23B (Genepharma) or sh‐NC (Genepharma) were subcutaneously inoculated into the right flank of mice ( n = 6 per group; 2 × 10 6 cells per mouse).

Techniques: Expressing

Journal: Thoracic Cancer

Article Title: Knockdown of circ‐RAD23B inhibits non‐small cell lung cancer progression via the miR ‐142‐3p/ MAP4K3 axis

doi: 10.1111/1759-7714.14319

Figure Lengend Snippet: Circ‐RAD23B knockdown blocked NSCLC development in vitro. (a) The expression of circ‐RAD23B in NSCLC tissues and normal tissues was measured by qPCR. (b) The expression of circ‐RAD23B in H1299, A549 and 16 HBE cells was detected by qPCR. (c) The efficiency of circ‐RAD23B interference was checked by qPCR. (d) The effect of circ‐RAD23B knockdown on cell viability was examined by CCK‐8 assay. (e) The effect of circ‐RAD23B knockdown on cell proliferation was examined by colony formation assay. (f and g) The effect of circ‐RAD23B knockdown on migration and invasion was examined by transwell assay. (h) Matrigel tube formation assay was performed to evaluate the angiogenic ability of HUVECs. (i) The effect of circ‐RAD23B knockdown on cell cycle distribution was determined by flow cytometry assay. (j) The effect of circ‐RAD23B knockdown on cell apoptosis was analyzed by flow cytometry assay. (k) Expression of PCNA and Bax affected by circ‐RAD23B knockdown was detected by western blot. * p < 0.05

Article Snippet: A549 cells infected with lentivirus sh‐circ‐RAD23B (Genepharma) or sh‐NC (Genepharma) were subcutaneously inoculated into the right flank of mice ( n = 6 per group; 2 × 10 6 cells per mouse).

Techniques: In Vitro, Expressing, CCK-8 Assay, Colony Assay, Migration, Transwell Assay, Tube Formation Assay, Flow Cytometry, Western Blot

Journal: Thoracic Cancer

Article Title: Knockdown of circ‐RAD23B inhibits non‐small cell lung cancer progression via the miR ‐142‐3p/ MAP4K3 axis

doi: 10.1111/1759-7714.14319

Figure Lengend Snippet: MiR‐142‐3p was a target of circ‐RAD23B. (a) the binding site between circ‐RAD23B and miR‐142‐3p was provided by Circinteractome tool. (b) The efficiency of miR‐142‐3p mimic was examined by qPCR. (c) Dual‐luciferase reporter assay was performed to validate the interaction between circ‐RAD23B and miR‐142‐3p. (d) The expression of miR‐142‐3p in NSCLC tissues and normal tissues was detected by qPCR. (e) The expression of miR‐142‐3p in H1299, A549 and 16 HBE cells was detected by qPCR. (f) The efficiency of miR‐142‐3p inhibitor was checked by qPCR. (g) The expression of miR‐142‐3p in H1299 and A549 cells transfected with si‐circ‐RAD23B‐1 or si‐circ‐RAD23B‐1 + miR‐142‐3p inhibitor was detected by qPCR. * p < 0.05

Article Snippet: A549 cells infected with lentivirus sh‐circ‐RAD23B (Genepharma) or sh‐NC (Genepharma) were subcutaneously inoculated into the right flank of mice ( n = 6 per group; 2 × 10 6 cells per mouse).

Techniques: Binding Assay, Luciferase, Reporter Assay, Expressing, Transfection

Journal: Thoracic Cancer

Article Title: Knockdown of circ‐RAD23B inhibits non‐small cell lung cancer progression via the miR ‐142‐3p/ MAP4K3 axis

doi: 10.1111/1759-7714.14319

Figure Lengend Snippet: Circ‐RAD23B knockdown blocked NSCLC development in vitro by mediating miR‐142‐3p. In H1299 and A549 cells transfected with si‐circ‐RAD23B‐1 or si‐circ‐RAD23B‐1 + miR‐142‐3p inhibitor, (a and b) cell proliferation was assessed by CCK‐8 assay and colony formation assay. (c and d) Cell migration and cell invasion were assessed by transwell assay. (e) The angiogenic ability was assessed by Matrigel tube formation assay. (f and g) Cell cycle distribution was determined by flow cytometry assay. (h) Cell apoptosis was analyzed by flow cytometry assay. (i and j) The expression of PCNA and Bax was measured by western blot. * p < 0.05

Article Snippet: A549 cells infected with lentivirus sh‐circ‐RAD23B (Genepharma) or sh‐NC (Genepharma) were subcutaneously inoculated into the right flank of mice ( n = 6 per group; 2 × 10 6 cells per mouse).

Techniques: In Vitro, Transfection, CCK-8 Assay, Colony Assay, Migration, Transwell Assay, Tube Formation Assay, Flow Cytometry, Expressing, Western Blot

Journal: Thoracic Cancer

Article Title: Knockdown of circ‐RAD23B inhibits non‐small cell lung cancer progression via the miR ‐142‐3p/ MAP4K3 axis

doi: 10.1111/1759-7714.14319

Figure Lengend Snippet: The circ‐RAD23B/miR‐142‐3p/MAP4K3 axis regulated the MAPK signaling pathway. (a) The protein level of MAP4K3 in H1299 and A549 cells transfected with si‐circ‐RAD23B‐1 or si‐circ‐RAD23B‐1 + miR‐142‐3p inhibitor was detected by western blot. (b and c) The protein levels of p‐ERK1/2, p‐JNK and p‐p38 in H1299 and A549 cells transfected with si‐circ‐RAD23B‐1 or si‐circ‐RAD23B‐1 + miR‐142‐3p inhibitor was detected by western blot. * p < 0.05

Article Snippet: A549 cells infected with lentivirus sh‐circ‐RAD23B (Genepharma) or sh‐NC (Genepharma) were subcutaneously inoculated into the right flank of mice ( n = 6 per group; 2 × 10 6 cells per mouse).

Techniques: Transfection, Western Blot

Journal: Thoracic Cancer

Article Title: Knockdown of circ‐RAD23B inhibits non‐small cell lung cancer progression via the miR ‐142‐3p/ MAP4K3 axis

doi: 10.1111/1759-7714.14319

Figure Lengend Snippet: Circ‐RAD23B knockdown inhibited tumor growth in vivo. In sh‐circ‐RAD23B or sh‐NC‐administered mice, (a) tumor volume was measured once a week. (b) Once tumor tissues were excised, tumor size was photographed, and tumor weight was measured. (c and d) The expression of circ‐RAD23B and miR‐142‐3p in the excised tumor tissues was detected by qPCR. (e) The expression of MAP4K3 in the excised tumor tissues was detected by western blot. * p < 0.05

Article Snippet: A549 cells infected with lentivirus sh‐circ‐RAD23B (Genepharma) or sh‐NC (Genepharma) were subcutaneously inoculated into the right flank of mice ( n = 6 per group; 2 × 10 6 cells per mouse).

Techniques: In Vivo, Expressing, Western Blot