Journal: Frontiers in Cell and Developmental Biology

Article Title: PD-L1 Is a Tumor Suppressor in Aggressive Endometrial Cancer Cells and Its Expression Is Regulated by miR-216a and lncRNA MEG3

doi: 10.3389/fcell.2020.598205

Figure Lengend Snippet: Loss of PD-L1 Induces Cell proliferation and Triggers EMT in Aggressive EC Cells. (A) Western blotting analysis of PD-L1 and MCL-1 expression in SPAC-1-L cells overexpressing PD-L1, and in PD-L1-silenced HEC-50 cells. (B) Cellular morphology of HEC-50 cells after knockdown of PD-L1. Scale bar, 100 μm. (C–E) Proliferation (C) , wound-healing (D) , and invasion (E) assays in EC cells after overexpression or knockdown of PD-L1. (F) A heatmap showing gene expression levels in human aggressive EC cell lines (Expression Atlas database). (G,H) Examination of gene expression in HEC-50 cells after knockdown of PD-L1 (G) and in SPAC-1-L cells after overexpression of PD-L1 (H) was performed using qRT-PCR assays. VIM, Vimentin. ∗ P < 0.05.

Article Snippet: The PD-L1 cDNA expresion vector pCMV6-PD-L1 (PD-L1-vec, RC213071), the MCL-1 cDNA expression vector pCMV6-MCL-1 (MCL-1-vec, RC200521), the MEG3 expression vector pCMV6-MEG3 (MEG3-vec, SC105816) and the pCMV6 control vector (Ctr-vec, PS100001) were purchased from OriGene (Rockville, MD, United States).

Techniques: Western Blot, Expressing, Over Expression, Quantitative RT-PCR

Journal: Frontiers in Cell and Developmental Biology

Article Title: PD-L1 Is a Tumor Suppressor in Aggressive Endometrial Cancer Cells and Its Expression Is Regulated by miR-216a and lncRNA MEG3

doi: 10.3389/fcell.2020.598205

Figure Lengend Snippet: PD-L1 Represses EMT by Decreasing MCL-1 Expression. (A) PD-L1-silenced HEC-50 cells or control cells were transfected with (or without) MCL-1 siRNA, and MCL-1 mRNA expression was verified using qRT-PCR analysis. (B) Cellular morphology of HEC-50 cells shown in panel (A) . Scale bar, 100 μm. (C) Wound-healing and invasion assays in HEC-50 cells transfected as indicated. (D) Western blotting analysis of the indicated proteins in HEC-50 cells transfected as indicated. (E) The IHC staining results were obtained from the HPA database. Immunohistochemical staining of EC tissues from the same patient showed a high MCL-1 and HSP47 expression and a low ZO-1 expression in EC tissues. (F) The correlation between the indicated genes and overall survival in patients with EC (KM plotter database). ∗ P < 0.05.

Article Snippet: The PD-L1 cDNA expresion vector pCMV6-PD-L1 (PD-L1-vec, RC213071), the MCL-1 cDNA expression vector pCMV6-MCL-1 (MCL-1-vec, RC200521), the MEG3 expression vector pCMV6-MEG3 (MEG3-vec, SC105816) and the pCMV6 control vector (Ctr-vec, PS100001) were purchased from OriGene (Rockville, MD, United States).

Techniques: Expressing, Transfection, Quantitative RT-PCR, Western Blot, Immunohistochemistry, Immunohistochemical staining, Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: PD-L1 Is a Tumor Suppressor in Aggressive Endometrial Cancer Cells and Its Expression Is Regulated by miR-216a and lncRNA MEG3

doi: 10.3389/fcell.2020.598205

Figure Lengend Snippet: Model Summarizing the Role of MEG3, MiR-216a, PD-L1, and MCL-1 in Controlling the Proliferative and Invasive Potential of aggressive EC cells. The downregulation of MEG3 causes an elevation of miR-216a expression. By targeting the 3′-UTR of PD-L1 mRNA, miR-216a suppresses PD-L1 expression. PD-L1 serves as a tumor-suppressor to inhibit the proliferation, EMT and invasive potential of aggressive EC cells via repressing MCL-1 expression.

Article Snippet: The PD-L1 cDNA expresion vector pCMV6-PD-L1 (PD-L1-vec, RC213071), the MCL-1 cDNA expression vector pCMV6-MCL-1 (MCL-1-vec, RC200521), the MEG3 expression vector pCMV6-MEG3 (MEG3-vec, SC105816) and the pCMV6 control vector (Ctr-vec, PS100001) were purchased from OriGene (Rockville, MD, United States).

Techniques: Expressing

Journal: BMC Cancer

Article Title: BIM EL is a key effector molecule in oxidative stress-mediated apoptosis in acute myeloid leukemia cells when combined with arsenic trioxide and buthionine sulfoximine

doi: 10.1186/1471-2407-14-27

Figure Lengend Snippet: BSO induces phosphorylation of BIM EL and MCL1 in mitochondria. HL60 cells were treated with ATO/BSO or ATO in the presence or absence of NAC or DTT for 12 h. A . The expression and phosphorylation of BIM were determined by immunoblotting. The lower panel indicates a longer exposure of the same blot. B, C . The expression and phosphorylation of BAD, BID, tBID, MCL1, BCLxL and BCL2 were determined by immunoblotting.

Article Snippet: The following antibodies were obtained from Cell Signaling Technology (Beverly, MA, USA): antibodies to the cleaved form of caspase 3 (C-cas3), caspase 9 (C-cas9), poly (ADP-ribose) polymerase (C-PARP); antibodies to normal and phosphorylated forms of MCL1 (MCL1, P-MCL1 at Ser159/Thr163), BCL2 (BCL2, P-BCL2 at Ser70), BIM (BIM, P-BIM at Ser69), JNK (JNK, P-JNK at Thr183/Tyr185), c-JUN (c-JUN, P-c-JUN at Ser63), p38 (p38, P-p38 at Thr180/Tyr182) and ERK1/2 (ERK1/2, P-ERK1/2 at Thr202/Tyr204); antibodies to actin, BAD, BID and BOK.

Techniques: Phospho-proteomics, Expressing, Western Blot

Journal: BMC Cancer

Article Title: BIM EL is a key effector molecule in oxidative stress-mediated apoptosis in acute myeloid leukemia cells when combined with arsenic trioxide and buthionine sulfoximine

doi: 10.1186/1471-2407-14-27

Figure Lengend Snippet: BSO induces the dissociation of phosphorylated BIM EL from MCL1 and the interaction with BAX. A and B , The dissociation of phosphorylated BIM EL and MCL1, and the interaction with BAX were determined by immunoprecipitation and immunoblotting with antibodies to their normal and phosphorylated forms. Values were normalized to actin or IgG(L), respectively and represent relative changes compared with control. A typical result of 3 independent experiments is shown. IP, immunoprecipitation; Ppt, immunoprecipitate; Sup, supernatant.

Article Snippet: The following antibodies were obtained from Cell Signaling Technology (Beverly, MA, USA): antibodies to the cleaved form of caspase 3 (C-cas3), caspase 9 (C-cas9), poly (ADP-ribose) polymerase (C-PARP); antibodies to normal and phosphorylated forms of MCL1 (MCL1, P-MCL1 at Ser159/Thr163), BCL2 (BCL2, P-BCL2 at Ser70), BIM (BIM, P-BIM at Ser69), JNK (JNK, P-JNK at Thr183/Tyr185), c-JUN (c-JUN, P-c-JUN at Ser63), p38 (p38, P-p38 at Thr180/Tyr182) and ERK1/2 (ERK1/2, P-ERK1/2 at Thr202/Tyr204); antibodies to actin, BAD, BID and BOK.

Techniques: Immunoprecipitation, Western Blot, Control

Journal: BMC Cancer

Article Title: BIM EL is a key effector molecule in oxidative stress-mediated apoptosis in acute myeloid leukemia cells when combined with arsenic trioxide and buthionine sulfoximine

doi: 10.1186/1471-2407-14-27

Figure Lengend Snippet: BSO triggers phosphorylation of MCL1and BIM EL via activation of JNK. A , The phosphorylation of JNK, ERK1/2 and p38 was determined by immunoblotting. HL60 cells were treated with ATO/BSO or ATO in the presence or absence of NAC or DTT for 12 h. B , The phosphorylation of BIM EL and MCL1, and the cleavage of caspase 3 and PARP, were determined by immunoblotting. HL60 cells were treated with ATO/BSO or ATO in the presence of SP600125 (10 μM) as a JNK inhibitor, U0126 (2 μM) as an ERK1/2 inhibitor, or SB203580 (10 μM) as a p38 inhibitor, PD035901 (100 nM)as a MEK1/2 inhibitor for 12 h. A typical result of 3 independent experiments is shown.

Article Snippet: The following antibodies were obtained from Cell Signaling Technology (Beverly, MA, USA): antibodies to the cleaved form of caspase 3 (C-cas3), caspase 9 (C-cas9), poly (ADP-ribose) polymerase (C-PARP); antibodies to normal and phosphorylated forms of MCL1 (MCL1, P-MCL1 at Ser159/Thr163), BCL2 (BCL2, P-BCL2 at Ser70), BIM (BIM, P-BIM at Ser69), JNK (JNK, P-JNK at Thr183/Tyr185), c-JUN (c-JUN, P-c-JUN at Ser63), p38 (p38, P-p38 at Thr180/Tyr182) and ERK1/2 (ERK1/2, P-ERK1/2 at Thr202/Tyr204); antibodies to actin, BAD, BID and BOK.

Techniques: Phospho-proteomics, Activation Assay, Western Blot

Journal: BMC Cancer

Article Title: BIM EL is a key effector molecule in oxidative stress-mediated apoptosis in acute myeloid leukemia cells when combined with arsenic trioxide and buthionine sulfoximine

doi: 10.1186/1471-2407-14-27

Figure Lengend Snippet: BSO triggers activation of ASK1 and JNK and induces phosphorylation of BIM EL and MCL1. A , The phosphorylation of ASK1 was determined by immunoblotting. HL60 cells were treated with ATO/BSO or ATO in the presence or absence of NAC or DTT for 12 h. B , The phosphorylation of JNK, BIM EL , and MCL1, and cleavage of caspase 3 and PARP, were determined by immunoblotting. HL60 cells were treated with ATO/BSO or ATO in the presence or absence of NQDI1 (10 μM) for 12 h. A typical result of 3 independent experiments is shown.

Article Snippet: The following antibodies were obtained from Cell Signaling Technology (Beverly, MA, USA): antibodies to the cleaved form of caspase 3 (C-cas3), caspase 9 (C-cas9), poly (ADP-ribose) polymerase (C-PARP); antibodies to normal and phosphorylated forms of MCL1 (MCL1, P-MCL1 at Ser159/Thr163), BCL2 (BCL2, P-BCL2 at Ser70), BIM (BIM, P-BIM at Ser69), JNK (JNK, P-JNK at Thr183/Tyr185), c-JUN (c-JUN, P-c-JUN at Ser63), p38 (p38, P-p38 at Thr180/Tyr182) and ERK1/2 (ERK1/2, P-ERK1/2 at Thr202/Tyr204); antibodies to actin, BAD, BID and BOK.

Techniques: Activation Assay, Phospho-proteomics, Western Blot