human normal lung epithelial cell line beas2b  (Procell Inc)

Journal: Molecular Therapy Oncolytics

Article Title: MicroRNA Modification of Coxsackievirus B3 Decreases Its Toxicity, while Retaining Oncolytic Potency against Lung Cancer

doi: 10.1016/j.omto.2020.01.002

Figure Lengend Snippet: miR-145 and miR-143 Are Significantly Downregulated In Lung Cancer Cells Compared with Normal Lung Epithelial Cells and Cardiomyocytes (A and B) Expression levels of miR-145 (A) and miR-143 (B) in human normal lung epithelial cells (BEAS2B and primary lung epithelial cells), cardiomyocytes (HL-1 mouse cardiomyocytes and human induced pluripotent stem cell-derived cardiomyocytes [iCMs]), KRAS mut non-small-cell lung cancer (NSCLC) cells (H2030, H23, and A549), TP53 mut /RB1 mut small-cell lung cancer (SCLC) cells (H524 and H526), and HeLa cells were measured by qRT-PCR and calculated using the equation RQ (relative quantity) = 2 −ΔΔCt . The results are presented as mean ± SD (n = 3). One-way ANOVA was used to assess the differences among different cell types. *p < 0.05, # p < 0.01, & p < 0.001. (C) Construction of miRNA-modified CVB3 (miR-CVB3). Four copies of miR-145 (5′-AAGGGATTCCTGGGAAAACTGGAC-3′) and two copies of miR-143 (5′-GAGCTACAGTGCTTCATCTCA-3′) target sequences were inserted between the 5′ UTR and VP4 of the CVB3 genome. A Kozak sequence was added before the start codon (ATG) of VP4 to facilitate the translation of viral protein in cancer cells.

Article Snippet: The KRAS mut lung adenocarcinoma cell lines of epithelial origin (i.e., A549 cells [#CCL-185], H2030 cells [#CRL-5914], and H23 cells [#CRL-5800]), the TP53 mut /RB1 mut SCLC epithelial cell lines (i.e., H524 cells [#CRL-5831] and H526 cells [#CRL-5811]), and the BEAS2B human normal lung epithelial cell line (#CRL-9609) were all obtained from the American Type Culture Collection.

Techniques: Expressing, Derivative Assay, Quantitative RT-PCR, Modification, Sequencing

Journal: Molecular Therapy Oncolytics

Article Title: MicroRNA Modification of Coxsackievirus B3 Decreases Its Toxicity, while Retaining Oncolytic Potency against Lung Cancer

doi: 10.1016/j.omto.2020.01.002

Figure Lengend Snippet: Both WT-CVB3 and miR-CVB3 Potently Destroy SCLC Cells, whereas Human Normal Lung Epithelial Cells Are Not Permissive to Either WT-CVB3 or miR-CVB3 (A–D) TP53 mut /RB1 mut SCLC cell lines (H524 and H526; A and B) and human normal lung epithelial cells (BEAS2B; C and D) were sham infected or inoculated with WT-CVB3 or miR-CVB3 at different MOIs as indicated for 72 h. Cell morphology was examined by light microscopy (original magnification, ×10; A and C). Cell viability was measured by the alamarBlue assay (mean ± SD, n = 3; B and D). An unpaired Student’s t test was used to assess the difference between cells treated with miR-CVB3 and WT-CVB3. *p < 0.05, # p < 0.01 compared to WT-CVB3.

Article Snippet: The KRAS mut lung adenocarcinoma cell lines of epithelial origin (i.e., A549 cells [#CCL-185], H2030 cells [#CRL-5914], and H23 cells [#CRL-5800]), the TP53 mut /RB1 mut SCLC epithelial cell lines (i.e., H524 cells [#CRL-5831] and H526 cells [#CRL-5811]), and the BEAS2B human normal lung epithelial cell line (#CRL-9609) were all obtained from the American Type Culture Collection.

Techniques: Infection, Light Microscopy, Alamar Blue Assay