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. 2020 Jun 18;5(12):e137112. doi: 10.1172/jci.insight.137112

Figure 5. Targeting MUC1-C in human colon cancer cells downregulates LGR5, BMI1, and stemness.

Figure 5

(A) Human SW620 colon cancer cells stably expressing a tet-CshRNA or tet-MUC1shRNA were treated with vehicle or 500 ng/mL DOX for 7 days. Lysates were immunoblotted with antibodies against the indicated proteins. (B) RNA-Seq was performed in triplicate on SW620 MUC1shRNA cells treated with vehicle or 500 ng/mL DOX for 7 days. Silencing MUC1-C expression was significantly associated with suppression of the indicated HALLMARK gene sets. (C) SW620 cells were left untreated or treated with 5 μM GO-203 for 48 hours. Lysates were immunoblotted with antibodies against the indicated proteins. (D) SW620 tet-MUC1shRNA cells treated with vehicle or 500 ng/mL DOX for 48 hours were monitored for wound healing in the scratch assay (left). The results are expressed as a percentage (mean ± SD of 3 biologic replicates) of the control at 0 hours (right). Scale bars: 100 μm. (E) SW620 cells left untreated or treated with 5 μM GO-203 for 48 hours were monitored for wound healing in the scratch assay (left). The results (mean ± SD of 3 biologic replicates) are expressed as a percentage of the control at 0 hours (right). Scale bars: 100 μm. (F) SW620 tet-CshRNA and SW620 tet-MUC1shRNA cells treated with vehicle or 500 ng/mL DOX for 24 hours were assayed for invasion (left). The results (mean ± SD of 3 biologic replicates) are expressed as the invasive cell number (right). Scale bar: 200 μm. (G) SW620 tet-CshRNA and SW620 tet-MUC1shRNA cells treated with vehicle or 500 ng/mL DOX for 7 days were assayed for colony formation (left). The results (mean ± SD of 3 biologic replicates) are expressed as colony number per field (right). (H) SW620 tet-CshRNA and SW620 tet-MUC1shRNA cells treated with vehicle or 500 ng/mL DOX for 5 days were assayed for tumorsphere formation (left). The results (mean ± SD of 3 biologic replicates) are expressed as tumorsphere number per field (right). Scale bar: 200 μm.