Journal: Journal of translational medicine

Article Title: MiR-30c-5p loss-induced PELI1 accumulation regulates cell proliferation and migration via activating PI3K/AKT pathway in papillary thyroid carcinoma.

doi: 10.1186/s12967-021-03226-1

Figure Lengend Snippet: Fig. 3 PELI1 activates the PI3K-AKT signaling pathways. A KEGG enrichment analysis showed that genes co-expressed with PELI1 are enriched in varied of pathway. B The protein levels of p-AKT, AKT, Ki-67 and MMP2 were assessed by western blotting in PELI1-overexpression or Peli1-silencing PTC cells. C Western blotting analysis showing the levels of PELI1 and p-AKT in the mouse tumor samples. D Representative IHC staining pictures of Ki-67 and MMP2 from mouse tumor sections (left panel), and the quantification of Ki-67 and MMP2 expression (right panel) (n = 3). The scale bars represent 50 µm. E Colony formation assay of PELI1-overexpression PTC cells after PI3K/AKT inhibition (LY294002; left panel), and the quantification was shown (right panel; n = 3). F Transwell assay was performed to estimate the cell migration of PELI1-overexpressing PTC cells after PI3K/AKT inhibition, the representative images (× 200) of Transwell assays were displayed (left panel), and the quantification was shown (right panel; n = 3). G Inhibition efficiency of PI3K/AKT and protein levels of Ki-67 and MMP2 in PELI1-overexpressing PTC cells after LY294002 treatment were determined by western blotting. The numbers are presented as fold increase over the LV-NC group. *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: Briefly, PTC tissue sections were incubated with an antibody to PELI1 (12,053–1- AP, diluted 1: 200; Proteintech, Rosemont, IL); mice tumor sections were incubated with an antibody to Ki-67 (ab16667, diluted 1: 200; Abcam, Cambridge, MA) or MMP2 (10,373–2-ap, diluted 1: 200; Proteintech) overnight at 4 °C, followed incubating by HPR-conjugated secondary antibody.

Techniques: Protein-Protein interactions, Western Blot, Over Expression, Immunohistochemistry, Expressing, Colony Assay, Inhibition, Transwell Assay, Migration

Journal: Journal of translational medicine

Article Title: MiR-30c-5p loss-induced PELI1 accumulation regulates cell proliferation and migration via activating PI3K/AKT pathway in papillary thyroid carcinoma.

doi: 10.1186/s12967-021-03226-1

Figure Lengend Snippet: Fig. 5 MiR-30c-5p inhibits PTC cell proliferation and migration by downregulating PELI1. A The cell proliferation of PTC cells with miR-30c-5p mimic or its control transfection was detected by CCK8 assay (n = 5). B Colony formation assays of PTC cells transfected with miR-NC or miR-30c-5p, and the quantification of colony formation was shown (n = 3). C Representative images of Transwell assays of PTC cells transfected with miR-30c-5p, and the quantification was shown (n = 3). D Representative images of scratch wound healing assays of PTC cells transfected with miR-30c-5p. E Transfection efficiency of increased PELI1 overexpression (oe-PELI1) was determined by western blot. F Colony formation analysis showed that oe-PELI1 could partially reverse the miR-30c-5p-mediated proliferation inhibition of PTC cells (n = 3). G Transwell analysis showed that oe-PELI1 significantly alleviated miR-30c-5p-mediated migration inhibition of PTC cells (n = 3). H p-AKT, AKT, Ki-67 and MMP2 protein levels were determined by western blot in each group. *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: Briefly, PTC tissue sections were incubated with an antibody to PELI1 (12,053–1- AP, diluted 1: 200; Proteintech, Rosemont, IL); mice tumor sections were incubated with an antibody to Ki-67 (ab16667, diluted 1: 200; Abcam, Cambridge, MA) or MMP2 (10,373–2-ap, diluted 1: 200; Proteintech) overnight at 4 °C, followed incubating by HPR-conjugated secondary antibody.

Techniques: Migration, Control, Transfection, CCK-8 Assay, Over Expression, Western Blot, Inhibition

Journal: Journal of translational medicine

Article Title: MiR-30c-5p loss-induced PELI1 accumulation regulates cell proliferation and migration via activating PI3K/AKT pathway in papillary thyroid carcinoma.

doi: 10.1186/s12967-021-03226-1

Figure Lengend Snippet: Fig. 6 EVs derived from miR-30c-5p-modified hUCMSC (miR-30c-5p-EVs) downregulate PELI1 expression in PTC cells. A miR-30c-5p-EVs and NC-EVs were observed under a transmission electron microscope (Scale bars: 200 nm), and the size distributions of these EVs were detected using the Nanoparticle Tracking Analysis. B Western blot analysis of TSG101 and HSP70 expression in miR-30c-5p-EVs and NC-EVs. Ponceau S staining served as a loading control. C The relative miR-30c-5p levels in miR-30c-5p-EVs and NC-EVs were detected by real-time PCR (n = 3). D Fluorescence was evaluated using laser confocal microscopy (Scale bars: 20 μm). E PTC cells treated with miR-30c-5p-EVs showed a significantly increased expression of miR-30c-5p in comparison with cells added with NC-EVs (n = 3). F, G Expression of PELI1 in PTC cells was assessed by real-time PCR (F) and Western blot (G). H p-AKT, AKT, Ki-67 and MMP2 protein levels were determined by western blot in each group. *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: Briefly, PTC tissue sections were incubated with an antibody to PELI1 (12,053–1- AP, diluted 1: 200; Proteintech, Rosemont, IL); mice tumor sections were incubated with an antibody to Ki-67 (ab16667, diluted 1: 200; Abcam, Cambridge, MA) or MMP2 (10,373–2-ap, diluted 1: 200; Proteintech) overnight at 4 °C, followed incubating by HPR-conjugated secondary antibody.

Techniques: Derivative Assay, Modification, Expressing, Transmission Assay, Microscopy, Western Blot, Staining, Control, Real-time Polymerase Chain Reaction, Fluorescence, Confocal Microscopy, Comparison

Journal: Journal of translational medicine

Article Title: MiR-30c-5p loss-induced PELI1 accumulation regulates cell proliferation and migration via activating PI3K/AKT pathway in papillary thyroid carcinoma.

doi: 10.1186/s12967-021-03226-1

Figure Lengend Snippet: Fig. 8 Proposed model of miR-30c-5p-EVs as new avenues for the treatment of PTC cancer. PELI1 was highly expressed in PTC cancer, while miR-30c-5p was poorly expressed. MiR-30c-5p could inhibit PTC cell proliferation and migration by negatively mediating the expression of the PELI1. MiR-30c-5p-EVs could significantly downregulate PELI1 expression and suppress the progression of PTC in vitro and in vivo, concomitant with reduced p-AKT, Ki-67 and MMP-2 expression

Article Snippet: Briefly, PTC tissue sections were incubated with an antibody to PELI1 (12,053–1- AP, diluted 1: 200; Proteintech, Rosemont, IL); mice tumor sections were incubated with an antibody to Ki-67 (ab16667, diluted 1: 200; Abcam, Cambridge, MA) or MMP2 (10,373–2-ap, diluted 1: 200; Proteintech) overnight at 4 °C, followed incubating by HPR-conjugated secondary antibody.

Techniques: Migration, Expressing, In Vitro, In Vivo