Polygonatum odoratum lectin promotes BECN1 expression and induces autophagy in malignant melanoma by regulation of miR1290

Figures & data

Figure 1 POL suppressed proliferation and induced autophagy in melanoma cells.

Notes: (A) Proliferative ability of A375 and HEMa-LP cells was measured by CCK8 assay after cells had been treated with POL. (B) Representative electron microscopy of A375 cells treated with 25 μg/mL POL for 24 hours. Arrows indicate autophagosomes. (C, D) Western blot assays show levels of LC3I, LC3II, and p62 following treatment with POL; GAPDH was used as a control. **P<0.01; ***P<0.001.
Abbreviation: POL, Polygonatum odoratum lectin.

Figure 2 POL increased levels of BECN1 mRNA and protein in melanoma cells.

Notes: (A) BECN1 levels were analyzed in 30 primary malignant melanoma tissues and adjacent normal tissues. (B, C) Expression of BECN1 mRNA in A375 cells treated with POL. (D, E) Western blot assays show levels of BECN1 following treatment with POL; GAPDH was used as a control. (F) Western blot identified LC3I, LC3II, and p62 protein-expression changes following treatment with POL alone or in combination with siBECN1. **P<0.01; ***P<0.001.
Abbreviation: POL, Polygonatum odoratum lectin.

Figure 3 POL may regulate BECN1 expression though miR1290.

Notes: (A) No significant change was detected in luciferase activity between the control group and the POL-treatment group. (B) Binding sites of miR1290 within the 3′-UTR of BECN1 were predicted by TargetScan. (C, D) Real-time PCR showed expression of miR1290 in POL-treated cells. (E) Real-time PCR showed that miR1290 mimics abolished the effect of POL on BECN1-mRNA expression. (F) Western blot showed that miR1290 mimics abolished the effect of POL on Becn1-protein expression. *P<0.05; **P<0.01; ***P<0.001.
Abbreviations: POL, Polygonatum odoratum lectin; PCR, polymerase chain reaction; NC, negative control.

Figure 4 miR1290 inhibited autophagy in melanoma cells by targeting BECN1.

Notes: (A) miR1290 levels were analyzed in 30 primary malignant melanoma tissues and adjacent normal tissues. (B) Pearson correlations of miR1290 and BECN1 expression in 30 malignant melanoma tissues were negative. (C) Transfection efficiency of miR1290 inhibitor was determined by PCR. (D) Representative electron microscopy of A375 cells after treatment with miR1290 inhibitor or in combination with siBECN1. Arrows indicate autophagosomes. (E) Overexpression of miR290 suppressed luciferase activity in A375 cells with the pMiR-BECN1-WT. (F) Real-time PCR showed that miR1290 downregulation led to an increase in the expression of endogenous BECN1. (G) Western blot identified Becn1, LC3I, LC3II, and P62 protein-expression changes following transfection with miR1290 alone or in combination with siBECN1. **P<0.01; ***P<0.001.
Abbreviations: PCR, polymerase chain reaction; NC, negative control; WT, wild type; Mut, mutant.

Figure 5 POL induces autophagy though regulating miR1290 in melanoma cells.

Notes: (A) Proliferative ability of A375 cells was measured by CCK-8 assay. (B) Representative electron microscopy of A375 cells. Arrows indicate autophagosomes. (C, D) Western blot shows Becn1, LC3I, LC3II, and P62 protein-expression changes; GAPDH was used as a control. *P<0.05; ***P<0.001.
Abbreviations: POL, Polygonatum odoratum lectin; NC, negative control.