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. 2020 Sep 20;20:132–146. doi: 10.1016/j.omto.2020.09.005

Figure 1.

Figure 1

MSC-Derived EVs Transfers miR-130b-3p to Lung Cancer Cells and miR-130b-3p Is Upregulated in Lung Cancer

(A) miRNA expression in lung cancer datasets GSE63805. (B) miRNA expression in lung cancer datasets GSE102286. Black represents miRNAs without significant difference, green represents downregulated miRNAs, and red represents upregulated miRNAs in lung cancer. (C) The intersection of upregulated miRNAs in lung cancer-related datasets and miRNA expression in MSC-EVs through the EVmiRNA database. The red and green circles represent the top 25 upregulated miRNAs in the lung cancer datasets GSE63805 and GSE102286. The center represents the overlapped miRNAs in 3 sets of data. (D) miR-130b-3p expression in lung cancer and adjacent normal tissues (n = 50) determined by qRT-PCR. ∗p < 0.05 compared to adjacent normal tissues. (E) Analysis of MSC osteogenic differentiation assessed by alkaline phosphatase(ALP) staining (200×), adipogenic differentiation detected by oil red O staining (400×), and chondrogenic differentiation detected by Alcian blue staining (400×). (F) EV morphology observed under TEM (representative bar, 100 nm). (G) EV diameters distribution determined by NTA. (H) EV markers determined by western blot analysis. (I) The uptake of EVs by H292 and H1299 cells observed by the fluorescence microscope (400×). (J) miR-130b-3p expression in H292 and H1299 cells after co-culture with MSC-EVs determined by qRT-PCR. ∗p < 0.05 compared to PBS treatment. (K) The transference of cy3-labeled miR-130b-3p mimic into H292 and H1299 cells observed by the fluorescence microscope (400×). (L) miR-130b-3p expression after the MSC condition medium was treated with RNase or RNase + Triton X-100 determined by qRT-PCR. ∗p < 0.05 compared to control. (M) miR-130b-3p expression after H292 and H1299 cells were treated with actinomycin D or MSC-EVs determined by qRT-PCR. ∗p < 0.05 compared to treatment with DMSO + EVs. The data were shown as means ± standard deviations. The cancer tissues were compared with adjacent normal tissues using the paired t test, while the other 2 groups were compared by the unpaired t test. The cell experiment was repeated 3 times.