Figure 5: IL-11 as a downstream target of TWF1.
From: MicroRNA-30c inhibits human breast tumour chemotherapy resistance by regulating TWF1 and IL-11
(a) Relative IL-11 expression levels measured by real-time PCR analyses. The miR-30c oligos (mimic) downregulated IL-11 expression, whereas the anti-miR-30c inhibitor upregulated IL-11 expression in MDA-MB-231 cells after 36 h of transfections, compared with the scrambled control (Scr). *P=0.03299, ***P=0.00052 (n=3) by a Student’s t-test. Error bars: s.d. (b) Overexpression of IL-11 reversed miR-30c-mediated sensitivity of MDA-MD-231 cells to doxorubicin (Doxo) and paclitaxel (Taxol) (n=6). Student’s t-test P=2.8E-05 (Doxo) and 1.1E-04 (Taxol), when comparing 30c/vector with 30c/IL-11. Error bars: s.d. (c) Survival of MDA-MB-231 cells incubated with the goat IgG control or a neutralizing antibody to human IL-11 (12 μg ml−1), upon 72 h exposure to 100 nM doxorubicin (Doxo). *****P=4.24E-06 by a Student’s t-test (n=6). Error bars: s.d. (d) Knockdown of IL-11 by siRNAs sensitized cytotoxicity of MDA-MB-231 cells to 72-h treatment of paclitaxel (Taxol, 10 nM). ****P=3.0E-05 by Student’s t-test (n=6). Error bars: s.d. (e) Reduction of IL-11 mRNA expression upon gene knockdowns of siTWF1 and siIL-11, compared with the scrambled control (Scr) (n=3), measured by real-time PCR. ***P=0.0009 (siTWF1) and 0.0003 (siIL-11) by a Student’s t-test. Error bars: s.d. (f) IL-11 levels detected in the cultured media of MDA-231 cells, upon transfections with scrambled, miR-30c and siRNAs for TWF1 or IL-11 (n=4). Student’s t-test P=0.0381 (30c), 0.0329 (siTWF1), 0.0007 (siIL11) compared to the scrambled control (Scr). Error bars: s.d. (g) Images of F-actin staining of MDA-MB-231 cells, transfected with siRNAs of IL-11 and scrambled control (Scr) (X1,000, green for F-actin staining and blue for DAPI staining), white scale bars, 5 μm. Cells were collected 36 h after transfections.
