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. 2017 Oct 26;9:387–398. doi: 10.1016/j.omtn.2017.10.016

Figure 2.

Figure 2

Antitumor Activity of Shrimp miR-34 in Breast Cancer Cells

(A) Sequence alignment of shrimp miR-34 and human miR-34a. (B) The expression of shrimp miR-34 in breast cancer cells. MDA-MB-231 or MDA-MB-435 cells were transfected with miR-34 or miR-34-scrambled. At 36 hr after transfection, the cells were subjected to northern blotting. U6 was used as a control. (C) Quantification of miR-34 in breast cancer cells. MDA-MB-231 or MDA-MB-435 cells were transfected with shrimp miR-34 or miR-34-scrambled. At 36 hr after transfection, the expression level of miR-34 in cells was analyzed by real-time qPCR. Negative control represents the cells without any treatment. (D) The effects of shrimp miR-34 on cancer cell proliferation. The proliferation rates of miR-34-transfected MDA-MB-231 and MDA-MB-435 cells were examined. MiR-34-scrambled and the cells without any treatment (negative control) were used as controls. (E) The influence of shrimp miR-34 on the cell cycle of breast cancer cells. MDA-MB-231 and MDA-MB-435 cells were transfected with the synthesized shrimp miR-34. As controls, the cells without any treatment (negative control) and the cells transfected with miR-34-scrambled were analyzed. At 36 hr after transfection, the cell cycle was examined by flow cytometry. (F) The impact of shrimp miR-34 on apoptosis of cancer cells. The cancer cells were transfected with shrimp miR-34 or miR-34-scrambled. Then 36 hr later, the percentage of apoptotic cancer cells was evaluated by flow cytometry. The cells without any treatment (negative control) were used as controls. (G) The influence of shrimp miR-34 expression on cancer cell metastasis. miR-34-transfected cancer cells were subjected to wound-healing assays to evaluate the migration ability of cancer cells. (H) The effects of shrimp miR-34 on breast cancer cell migration. MDA-MB-231 and MDA-MB-435 cells were transfected with synthesized miR-34 or miR-34-scrambled and further cultured for 36 hr. (I) The effects of shrimp miR-34 expression on breast cancer cell adhesion. As a control, miR-34-scrambled was included in the assay. (J) The role of shrimp miR-34 in breast cancer cell invasion. Scale bar, 100 μm. In all panels, statistically significant differences between treatments are represented with asterisks (error bar, SD; *p < 0.05 and **p < 0.01).