Sinomenine restrains breast cancer cells proliferation, migration and invasion via modulation of miR-29/PDCD-4 axis

Figures & data

Figure 1. The chemical structure of Sino. The molecular formula of Sinomenine (Sino) is C₁₉H₂₃NO₄, and the molecular weight is 329.38.

Figure 2. Impacts of Sino on MDA-MB-231 cells growth. Diverse dosages of Sino (0, 1, 2, 4, 8 and 16 μM) were exploited for the stimulation of MDA-MB-231 cells. (A) Diagram showed the CCK-8 analytical results of cell viability after Sino stimulation. (B) BrdU experiment was adopted for the detection of cell proliferation after management with 4 μM of Sino. (C and D) Western blot experiment was enforced for the determination of PCNA, CyclinD1, CDK4 and p16 in Sino-processed cells. (E) Flow cytometry experiment was executed for the estimation of cell apoptosis in Sino-processed cells. (F) Western blot experiment was worked out for the examination of pro-Caspase-3/-9 and cleaved-Caspase-3/-9 in Sino-managed cells. *p < .05; **p < .01; ***p < .001.

Figure 3. Impacts of Sino on MDA-MB-231 cells migration and invasion. The 4 μM Sino was employed for the administration of MDA-MB-231 cells, and subsequently Transwell experiment was applied for the evaluation of (A) cell migration and (B) cell invasion in MDA-MB-231 cells after Sino manipulation. *p < .05.

Figure 4. Impacts of Sino on the expression of miR-29 in MDA-MB-231 cells. The different dosages of Sino (0, 1, 2, and 4 μM) were utilized to manage MDA-MB-231 cells. qRT-PCR experiment was implemented for the examination of miR-29 expression in MDA-MB-231 cells after Sino stimulation. *p < .05; ***p < .001.

Figure 5. Impacts of miR-29 on cell growth, migration and invasion in Sino-processed MDA-MB-231 cells. After miR-29 mimic, miR-29 inhibitor and the corresponding NC transfection, (A) miR-29 expression was assessed by qRT-PCR experiment. (B) The positive BrdU cells, (C and D) PCNA, CyclinD1, CDK4 and p16, (E) cell apoptosis, (F) pro-Caspase-3/-9 and cleaved-Caspase-3/-9, (G) cell migration and (H) cell invasion were respectively evaluated by BrdU, flow cytometry, western blot and Transwell trials in Sino-managed and miR-29 mimic/inhibitor transfected MDA-MB-231 cells. *p < .05; **p < .01; ***p < .001 vs NC or Control group; #p < .05; ##p < .01; ###p < .0.01 vs Sino+NC group.

Figure 6. Impacts of PDCD-4 on JNK and MEK/ERK pathways in MDA-MB-231 cells. (A) PDCD-4 expression in Sino-managed and miR-29 mimic/inhibitor transfected MDA-MB-231 cells was evaluated through applying western blot and qRT-PCR experiments. The pc-PDCD-4, sh-PDCD-4 and the correlative controls were transfected into MDA-MB-231 cells, (B) PDCD-4 expression was measured by western blot ans qRT-PCR experiments; (C) p/t-JNK, and (D) p/t-MEK and p/t-ERK protein levels were then appraised via adopting western blot experiment. *p < .05; **p < .01; ***p < .001.

Data availability statement

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.