Journal: Cancer Science

Article Title: The Positive Feedback of lncRNA ANRIL / miRNA ‐339‐5p/ ZBTB7A Suppresses Metastasis of Nasopharyngeal Carcinoma Cells via SREBP1 ‐ FASN

doi: 10.1111/cas.70225

Figure Lengend Snippet: Expressions of ZBTB7A and relative genes in NPC sublines with stable transfection of shZBTB7A and ANRIL. (A) Representative WB images of ZBTB7A, SREBP1, and FASN expressions in NPC sublines with stable knockdown of ZBTB7A and overexpression of ANRIL. (B) Protein levels of ZBTB7A, SREBP1, and FASN in the sublines. (C) The RNA levels of ZBTB7A, SREBF1, FASN, and ANRIL in the sublines. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Cell lysates were incubated with antibodies against ZBTB7A, SREBP1, FASN (Santa Cruz, USA) or control IgG (ABclonal Biotechnology, China) overnight at 4°C.

Techniques: Stable Transfection, Knockdown, Over Expression

Journal: Cancer Science

Article Title: The Positive Feedback of lncRNA ANRIL / miRNA ‐339‐5p/ ZBTB7A Suppresses Metastasis of Nasopharyngeal Carcinoma Cells via SREBP1 ‐ FASN

doi: 10.1111/cas.70225

Figure Lengend Snippet: Metastatic ability of NPC sublines is reduced in mice. (A) The figure illustrates the establishment of an inguinal lymph node metastasis model in mice. A representative photograph of a primary foot‐pad tumor (black arrow) and inguinal lymph node (red arrow) captured at 7 weeks post‐transplantation. (B) The photograph and average volumes of the inguinal lymph nodes in the CtrlANRIL‐5‐8F‐shZBTB7A + DMSO or FAT, ANRIL‐5‐8F‐shZBTB7A + DMSO or FAT groups ( n = 5 in each group). (C) Representative images of the foot‐pad tumor and inguinal lymph node tissues with H&E staining from the groups are shown. The formations of foot‐pad tumor or lymphatic metastases are indicated (black arrows or red arrows); none in the shZBTB7A‐ANRIL+FAT group (Short scale bar = 200 μm, Long scale bar = 50 μm). (D) Representative images of IHC analysis in SREBP1 and FASN expressions of the tissues of the groups (Short scale bar = 200 μm, Long scale bar = 50 μm). (E) The bar charts represent AOD values of SREBP1 and FASN in the tissues of the groups. (F) Summary of lymphatic metastasis in the groups. * p < 0.05, ** p < 0.01, *** p < 0.001; ns , not significant.

Article Snippet: Cell lysates were incubated with antibodies against ZBTB7A, SREBP1, FASN (Santa Cruz, USA) or control IgG (ABclonal Biotechnology, China) overnight at 4°C.

Techniques: Transplantation Assay, Staining

Journal: Cancer Science

Article Title: The Positive Feedback of lncRNA ANRIL / miRNA ‐339‐5p/ ZBTB7A Suppresses Metastasis of Nasopharyngeal Carcinoma Cells via SREBP1 ‐ FASN

doi: 10.1111/cas.70225

Figure Lengend Snippet: Lipid metabolism of NPC sublines is diminished. (A) HK1 and 5‐8F sublines were stained with LDs fluorescence, with OA used as a positive control (Scale bar = 200 μm), along with the quantification of the results. (B) The concentrations of TG and TC in HK1 and 5‐8F sublines were determined. (C) The concentrations of SREBP1 and FASN in the cells with stable dual‐gene transfections were measured. (D) Representative photographs of LDs in the cells were captured by TEM (Scale bar = 2 μm; LDs: Red arrows). The number of LDs in the cells was analyzed. (E) Representative images of mitochondria in the cells (Long scale bar = 2 μm, Short scale bar = 200 nm; Red arrows represent the representative mitochondria). DMSO or FAT were added to the assays, respectively (B–E). * p < 0.05, ** p < 0.01, *** p < 0.001; ns , not significant.

Article Snippet: Cell lysates were incubated with antibodies against ZBTB7A, SREBP1, FASN (Santa Cruz, USA) or control IgG (ABclonal Biotechnology, China) overnight at 4°C.

Techniques: Staining, Fluorescence, Positive Control, Transfection

Journal: Cancer Science

Article Title: The Positive Feedback of lncRNA ANRIL / miRNA ‐339‐5p/ ZBTB7A Suppresses Metastasis of Nasopharyngeal Carcinoma Cells via SREBP1 ‐ FASN

doi: 10.1111/cas.70225

Figure Lengend Snippet: ANRIL/miR‐339‐5p/ZBTB7A axis regulates SREBF1‐FASN. (A) MiR‐339‐5p binding sites on ANRIL and ZBTB7A were predicted. (B) Luciferase activities of HK1 and 5‐8F cells co‐transfected with ANRIL WT/MUT or ZBTB7A WT/MUT and miR‐339‐5p mimic or mimic NC were respectively analyzed. (C) A potential interaction was predicted between ZBTB7A and the promoter region of ANRIL and (D) luciferase activities of HK1 and 5‐8F cells co‐transfected with ANRIL WT/MUT and ZBTB7A or NC‐ZBTB7A. (E) ChIP‐qPCR was used to analyze whether ZBTB7A is directly bound to the ANRIL promoter. (F) Potential interactions between ZBTB7A and the promoter region of SREBP1 or FASN were predicted. (G) Luciferase activities of HK1 and 5‐8F cells co‐transfected with SREBF1 WT/MUT or FASN WT/MUT and ZBTB7A or NC‐ZBTB7A. (H) ChIP‐qPCR was used to analyze whether ZBTB7A is directly bound to the SREBF1 or FASN promoter. * p < 0.05, ** p < 0.01, *** p < 0.001; ns , not significant.

Article Snippet: Cell lysates were incubated with antibodies against ZBTB7A, SREBP1, FASN (Santa Cruz, USA) or control IgG (ABclonal Biotechnology, China) overnight at 4°C.

Techniques: Binding Assay, Luciferase, Transfection, ChIP-qPCR

Journal: Cancer Science

Article Title: The Positive Feedback of lncRNA ANRIL / miRNA ‐339‐5p/ ZBTB7A Suppresses Metastasis of Nasopharyngeal Carcinoma Cells via SREBP1 ‐ FASN

doi: 10.1111/cas.70225

Figure Lengend Snippet: The mechanism of positive feedback of ANRIL/microRNA‐339‐5p/ZBTB7A suppressing NPC metastasis. ZBTB7A knockdown weakens ANRIL absorption of miR‐339‐5p, while ANRIL overexpression reinforces ZBTB7A suppression of SREBP1 and FASN.

Article Snippet: Cell lysates were incubated with antibodies against ZBTB7A, SREBP1, FASN (Santa Cruz, USA) or control IgG (ABclonal Biotechnology, China) overnight at 4°C.

Techniques: Knockdown, Over Expression

Journal: Journal of Inflammation Research

Article Title: Qingyi Decoction Alleviates Alcoholic Pancreatitis by Improving Glycerolipid Homeostasis via the AMPK/SREBP-1c/PPARα Pathway

doi: 10.2147/JIR.S537259

Figure Lengend Snippet: ( A–E ) The mRNA levels of SREBP-1c, ACC1, FASN, PPARα, and CPT1A in pancreatic tissues measured by RT-qPCR (n=3, * P <0.05, ** P <0.01, *** P <0.001, ns indicates no statistical significance).

Article Snippet: Subsequently, membranes were blocked with a 5% skim milk solution for 1.5 h. Antibodies AMPKα (Rabbit, Proteintech, 1:1000), p-AMPKα (Rabbit, CST, 1:1000), SREBP-1c (Rabbit, Affinity, 1:1000), ACC1 (Rabbit, ABclonal, 1:1000), FASN (Rabbit, Proteintech, 1:5000), PPARα (Rabbit, Proteintech, 1:1000) and CPT1A (Mouse, Affinity, 1:5000) were added overnight at 4°C, and horseradish peroxidase (HRP) conjugated secondary antibodies (ABclonal, 1:10000) was incubated for 1 h at room temperature.

Techniques: Quantitative RT-PCR

Journal: Journal of Inflammation Research

Article Title: Qingyi Decoction Alleviates Alcoholic Pancreatitis by Improving Glycerolipid Homeostasis via the AMPK/SREBP-1c/PPARα Pathway

doi: 10.2147/JIR.S537259

Figure Lengend Snippet: ( A–F ) The protein levels of AMPKα, p-AMPKα, SREBP-1c, ACC1, FASN, PPARα, and CPT1A in pancreatic tissues detected by Western blot (n=3, * P <0.05, ** P <0.01, *** P <0.001, ns indicates no statistical significance).

Article Snippet: Subsequently, membranes were blocked with a 5% skim milk solution for 1.5 h. Antibodies AMPKα (Rabbit, Proteintech, 1:1000), p-AMPKα (Rabbit, CST, 1:1000), SREBP-1c (Rabbit, Affinity, 1:1000), ACC1 (Rabbit, ABclonal, 1:1000), FASN (Rabbit, Proteintech, 1:5000), PPARα (Rabbit, Proteintech, 1:1000) and CPT1A (Mouse, Affinity, 1:5000) were added overnight at 4°C, and horseradish peroxidase (HRP) conjugated secondary antibodies (ABclonal, 1:10000) was incubated for 1 h at room temperature.

Techniques: Western Blot

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Platelet-Derived Growth Factor C Facilitates Malignant Behavior of Pancreatic Ductal Adenocarcinoma by Regulating SREBP1 Mediated Lipid Metabolism.

doi: 10.1002/advs.202407069

Figure Lengend Snippet: Figure 5. PDGFC regulates SREBP1 via PI3K/AKT signaling pathway to promote lipid biosynthesis. A) GO molecular function enrichment of DEGs in BxPC-3 cells silencing PDGFC or not. B) The mRNA level of SREBF1 between PDAC tumor tissue or normal tissue in GEPIA2 database. C) The mRNA level of SREBF1 after PDGFC silencing in SW1990 cells. D) KEGG enrichment of DEGs after PDGFC silencing or not. E) GSEA enrichment results suggest PI3K/AKT signaling pathway is the downstream target of PDGFC. F) The protein level of PDGFR/PI3K/AKT signaling pathway and SREBP1 after PDGFC silencing were confirmed by western blotting. G) The mRNA level of SREBF1 after capivasertib treatment in PDAC cells. H) The protein level

Article Snippet: To further elucidate the pivotal role of PI3K/AKT signaling pathway in PDGFC regulation of SREBP1, we treated PDAC cells with capivasertib, a specific AKT inhibitor (TargetMol, Shanghai, China).

Techniques: Western Blot

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Platelet-Derived Growth Factor C Facilitates Malignant Behavior of Pancreatic Ductal Adenocarcinoma by Regulating SREBP1 Mediated Lipid Metabolism.

doi: 10.1002/advs.202407069

Figure Lengend Snippet: Figure 6. PDGFC facilitates PDAC progression by upregulating SREBP1 expression. A) The mRNA and protein level after SREBF1 overexpression in BxPC-3 and SW1990 cells. B) Cell growth curve after SREBF1 overexpression in PDAC cells. C) Cell migration and invasion ability after SREBF1 overex- pression in BxPC-3 cells. D) Cell migration and invasion ability after SREBF1 overexpression in SW1990 cells. E) The mRNA level after PDGFC silencing and PDGFC silencing with SREBF1 overexpression confirmed by RT-qPCR. F) The protein level after PDGFC silencing and PDGFC silencing with SREBF1 overexpression confirmed by western blotting. G) Cell growth curve after PDGFC silencing and PDGFC silencing with SREBF1 overexpression. H) Sta- tistical analysis of cell migration and invasion ability. I) Representative images of cell migration and invasion ability in BxPC-3 and SW1990 cells. Data are presented as mean ± SD (n = 3). *p < 0.05; **p < 0.01; ***p < 0.001 according to Student’s t-test.

Article Snippet: To further elucidate the pivotal role of PI3K/AKT signaling pathway in PDGFC regulation of SREBP1, we treated PDAC cells with capivasertib, a specific AKT inhibitor (TargetMol, Shanghai, China).

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