sw48 colon cancer cell lines  (ATCC)

Journal: Molecular Therapy Oncolytics

Article Title: M2 macrophage-derived exosomal microRNA-155-5p promotes the immune escape of colon cancer by downregulating ZC3H12B

doi: 10.1016/j.omto.2021.02.005

Figure Lengend Snippet: Colon cancer progression was affected by M2 macrophage-derived exosomes (A) Immunostaining of expression of M2 macrophage markers (CD68, CD163, and CD206) (×200). (B) Western blot analysis of protein expression patterns of the M2 macrophage markers (CD68, CD163, and CD206). (C) The effect of M2 macrophages on SW48 cell proliferation detected by EdU assay. (D) The effect of M2 macrophages on SW48 cell apoptosis rate measured by flow cytometry. (E) Nanoparticle tracking analysis of particle size of exosomes. (F) The ultrastructure of the exosomes (×5,000) observed under a transmission electron microscope. (G) Western blot analysis of the expression of exosome markers (CD63 and CD81). Control, the supernatant after exosome isolation. Exosome, extracted exosomes. (H) Internalization of M2 macrophage-derived exosomes in SW48 cells (×400). (I) SW48 cell proliferation measured by EdU assay upon coculture with GW4869-treated M2 macrophages. (J) Western blot analysis of exosome-specific marker proteins CD63 and CD81 in SW48 cells cocultured with GW4869-treated M2 macrophages. The data are all measurement data expressed as mean ± standard deviation and compared by independent-sample t test. ∗p < 0.05. The cell experiment was repeated three times.

Article Snippet: Colon cancer cell line (SW48) cells and human normal colonic epithelial cells (CCD841CoN) were procured from American Type Culture Collection (Rockville, MD, USA).

Techniques: Derivative Assay, Immunostaining, Expressing, Western Blot, EdU Assay, Flow Cytometry, Transmission Assay, Microscopy, Control, Isolation, Marker, Standard Deviation

Journal: Molecular Therapy Oncolytics

Article Title: M2 macrophage-derived exosomal microRNA-155-5p promotes the immune escape of colon cancer by downregulating ZC3H12B

doi: 10.1016/j.omto.2021.02.005

Figure Lengend Snippet: M2 macrophage-derived exosomes transferred miR-155-5p to promote proliferation and to repress apoptosis of SW48 cells (A) qRT-PCR examining the expression patterns of miR-155-5p in M2 macrophage-derived exosomes compared with M1 macrophage-derived exosomes (normalized to U6). (B) Immunofluorescence demonstrating transferring of miR-155-5p from M2 macrophages to SW48 cells (scale bars, 25 μm). (C) qRT-PCR examining the expression patterns of miR-155-5p in exo-miR-155-5p inhibitor and exo-inhibitor NC (normalized to U6). (D) qRT-PCR analysis of miR-155-5p in SW48 cells after coculture with exo-miR-155-5p inhibitor and exo-inhibitor NC (normalized to U6). (E) SW48 cell proliferation after coculture with exo-miR-155-5p inhibitor and exo-inhibitor NC detected by EdU assay. (F) SW48 cell apoptosis after coculture with exo-miR-155-5p inhibitor and exo-inhibitor NC detected by flow cytometry. The data are measurement data expressed as mean ± standard deviation. Comparisons between two groups were analyzed by independent-sample t test. Comparisons among multiple groups were analyzed by one-way ANOVA, followed by Tukey’s post hoc test. ∗p < 0.05. The cell experiment was repeated three times.

Article Snippet: Colon cancer cell line (SW48) cells and human normal colonic epithelial cells (CCD841CoN) were procured from American Type Culture Collection (Rockville, MD, USA).

Techniques: Derivative Assay, Quantitative RT-PCR, Expressing, Immunofluorescence, Transferring, EdU Assay, Flow Cytometry, Standard Deviation

Journal: Molecular Therapy Oncolytics

Article Title: M2 macrophage-derived exosomal microRNA-155-5p promotes the immune escape of colon cancer by downregulating ZC3H12B

doi: 10.1016/j.omto.2021.02.005

Figure Lengend Snippet: miR-155-5p negatively targeted ZC3H12B (A) The online site prediction of the binding site of miR-155-5p to ZC3H12B. (B) GEPIA website prediction of ZC3H12B expression patterns in colon cancer. (C) qRT-PCR examining the mRNA expression patterns of ZC3H12B in colon cancer and adjacent normal tissue samples (n = 36; normalized to GAPDH). (D) qRT-PCR examining the mRNA expression patterns of ZC3H12B in SW48 colon cancer cells and CCD841CoN human normal colon epithelial cells (normalized to GAPDH). (E) Dual luciferase reporter assay verifying the targeted binding of miR-155-5p to ZC3H12B. (F) qRT-PCR and western blot analysis examining the mRNA and protein expression patterns of ZC3H12B in SW48 cells after alteration of miR-155-5p (normalized to GAPDH). The data are measurement data expressed as mean ± standard deviation. Comparisons between two groups were analyzed by unpaired t test. Data were compared between cancer tissues and adjacent normal tissues by paired t test. Comparisons among multiple groups were analyzed by one-way ANOVA, followed by Tukey’s post hoc test. ∗p < 0.05. The cell experiment was repeated three times.

Article Snippet: Colon cancer cell line (SW48) cells and human normal colonic epithelial cells (CCD841CoN) were procured from American Type Culture Collection (Rockville, MD, USA).

Techniques: Binding Assay, Expressing, Quantitative RT-PCR, Luciferase, Reporter Assay, Western Blot, Standard Deviation

Journal: Molecular Therapy Oncolytics

Article Title: M2 macrophage-derived exosomal microRNA-155-5p promotes the immune escape of colon cancer by downregulating ZC3H12B

doi: 10.1016/j.omto.2021.02.005

Figure Lengend Snippet: ZC3H12B downregulated IL-6 expression to repress immune escape (A) ZC3H12B and IL-6 mRNA expression patterns after alteration of ZC3H12B detected by qRT-PCR. (B) ZC3H12B and IL-6 protein expression patterns after alteration of ZC3H12B detected by western blot analysis. (C) Luciferase reporter gene system validating the binding of ZC3H12B and IL-6 mRNA 3¢ UTR. (D) The residual amount of IL-6 mRNA at different time points in SW48 cells treated with sh-ZC3H12B and actinomycin D (Act D) measured by qRT-PCR (normalized to GAPDH). (E) Western blot analysis of ZC3H12B and IL-6 protein expression patterns in SW48 cells after overexpression of ZC3H12B and IL-6 (normalized to GAPDH). (F) ELISA examining IL-6 expression patterns in the supernatant of SW48 cells after overexpression of ZC3H12B and IL-6. (G) Flow cytometry examining the proliferation of CD3 + T cells and the proportion of activated IFN-γ + T cells after coculture of SW48 colon cancer cells treated with oe-ZC3H12B and oe-IL-6. The data are measurement data expressed as mean ± standard deviation. Comparisons between two groups were analyzed by independent-sample t test. Comparisons among multiple groups were analyzed by one-way ANOVA, followed by Tukey’s post hoc test. ∗p < 0.05. #p < 0.05. The cell experiment was repeated three times.

Article Snippet: Colon cancer cell line (SW48) cells and human normal colonic epithelial cells (CCD841CoN) were procured from American Type Culture Collection (Rockville, MD, USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Luciferase, Binding Assay, Over Expression, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Standard Deviation

Journal: Molecular Therapy Oncolytics

Article Title: M2 macrophage-derived exosomal microRNA-155-5p promotes the immune escape of colon cancer by downregulating ZC3H12B

doi: 10.1016/j.omto.2021.02.005

Figure Lengend Snippet: miR-155-5p promoted immune escape of colon cancer by downregulating the expression of ZC3H12B in vitro SW48 cells were transfected with oe-NC, miR-155-5p mimic + oe-NC, oe-ZC3H12B, or miR-155-5p mimic + oe-ZC3H12B. (A) qRT-PCR examining the expression patterns of miR-155-5p expression and ZC3H12B mRNA in SW48 cells. (B) Western blot analysis examining the protein expression patterns of ZC3H12B and IL-6 in SW48 cells. (C) ELISA examining the expression patterns of IL-6 in the supernatant of SW48 cells. (D) Flow cytometry examining the proliferation of T cells and the proportion of activated INF-γ + T cells after coculture of SW48 cells with different transfections. (E) qRT-PCR examining the expression patterns of miR-155-5p and ZC3H12B mRNA in SW48 cells after coculture with exo-miR-155-5p inhibitor (SW48 cells were treated with PBS as control, the same as below). (F) Western blot analysis examining the expression patterns of ZC3H12B and IL-6 protein in SW48 cells after coculture with exo-miR-155-5p inhibitor. (G) ELISA examining the expression patterns of IL-6 in the supernatant of SW48 cells after coculture with exo-miR-155-5p inhibitor. (H) Flow cytometry examining the proliferation of T cells and the proportion of activated IFN-γ + T cells after coculture with exo-miR-155-5p inhibitor-treated SW48 cells. The data are measurement data expressed as mean ± standard deviation. Comparisons between two groups were analyzed by independent-sample t test. Comparisons among multiple groups were analyzed by one-way ANOVA, followed by Tukey’s post hoc test. ∗p < 0.05. The cell experiment was repeated three times.

Article Snippet: Colon cancer cell line (SW48) cells and human normal colonic epithelial cells (CCD841CoN) were procured from American Type Culture Collection (Rockville, MD, USA).

Techniques: Expressing, In Vitro, Transfection, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Control, Standard Deviation

Journal: Molecular Therapy Oncolytics

Article Title: M2 macrophage-derived exosomal microRNA-155-5p promotes the immune escape of colon cancer by downregulating ZC3H12B

doi: 10.1016/j.omto.2021.02.005

Figure Lengend Snippet: miR-155-5p delivered by M2 macrophage-derived exosomes diminished ZC3H12B expression to accelerate immune escape in colon cancer in vivo Mice were injected with SW48 cells treated with exo-inhibitor-NC + oe-NC, exo-miR-155-5p inhibitor + oe-NC, exo-inhibitor-NC + oe-ZC3H12B, or exo-miR-155-5p inhibitor + oe-ZC3H12B. (A) Expression patterns of M2 macrophage-derived exosomes analyzed by transmission electron microscope and immunofluorescence assay in mouse tumor tissues. (B) qRT-PCR examining the expression patterns of miR-155-5p expression and ZC3H12B mRNA in mouse tumor tissues. (C) Tumor growth of mice. (D) Tumor volume of mice. (E) Tumor weight of mice. (F) ELISA examining the expression patterns of IL-6 in spleen cell lysates of mice. (G) The expression patterns of ZC3H12B and IL-6 protein in tumor tissues of mice without any other treatments determined by immunohistochemistry. (H) miR-155-5p expression patterns in tumor tissues of mice without any other treatments. (I) Flow cytometry examining T cells in spleen cells of mice without any other treatments. The data are measurement data expressed as mean ± standard deviation. Comparisons between two groups were analyzed by independent-sample t test. Comparisons among multiple groups were analyzed by one-way ANOVA, followed by Tukey’s post hoc test. ∗p < 0.05. n = 5.

Article Snippet: Colon cancer cell line (SW48) cells and human normal colonic epithelial cells (CCD841CoN) were procured from American Type Culture Collection (Rockville, MD, USA).

Techniques: Derivative Assay, Expressing, In Vivo, Injection, Transmission Assay, Microscopy, Immunofluorescence, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Flow Cytometry, Standard Deviation

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Triple blockade of EGFR, MEK and PD-L1 has antitumor activity in colorectal cancer models with constitutive activation of MAPK signaling and PD-L1 overexpression

doi: 10.1186/s13046-019-1497-0

Figure Lengend Snippet: CMS4 gene expression signature in a preclinical model of colon cancer cells with acquired resistance to MEKi. Agilent microarray analyses were performed to assess baseline gene expression profile for SW48 and SW48-MR Colon cancer cell lines, as described in Materials and Methods. (Log) fold change indicates relative microRNA expression levels in SW48-MR respect to SW48 cells. The genes whose expression is up regulated in SW48-MR compared to SW48 cells are represented in schematic tables. These genes are involved in CMS4 gene signature

Article Snippet: The human SW48 (catalogue number: HTL99020) and HCT116 (catalogue number: HTL99017) colon cancer cell lines were obtained from IRCCS “Azienda Ospedaliera Universitaria San Martino-IST Istituto Nazionale per la Ricerca sul Cancro” (Genova, Italy).

Techniques: Gene Expression, Microarray, Expressing