Journal: Frontiers in Oncology

Article Title: Kiss1 Inhibits the Proliferation of Nasopharyngeal Carcinoma Cells Via Activation of the LKB1/AMPK Pathway

doi: 10.3389/fonc.2021.724251

Figure Lengend Snippet: Overexpression of KISS1 increased LKB1, phosphorylation of LKB1 and AMPKα. (A) Schematic diagram of AMPK signaling pathway. (B) Heat map of gene expression related to AMPK pathway in 5-8F-vehicle cells, 5-8F-KISS1 cells, 5-8F-1R cells and 5-8F-1R-KISS1 cells. (C) RT-PCR verified the expression of LKB1, CAMKK1, CAMKK2, AMPKα1, AMPKα2 genes in 5-8F-vehicle cells, 5-8F-KISS1 cells, 5-8F-1R cells and 5-8F-1R-KISS1 cells. (D) Western blot analysis of LKB1, P-LKB1, CAMMKK2, AMPKα, P-AMPKα and GAPDH expression in 5-8F-vehicle cells, 5-8F-KISS1 cells, 5-8F-1R cells and 5-8F-1R-KISS1 cells. (E) Representative bands show that the expression of LKB1, P-LKB1, CAMKK2, AMPKα, P-AMPKα and GAPDH in 5-8F-vehicle cells, 5-8F-KISS1 cells, 5-8F-1R cells and 5-8F-1R-KISS1 cells. (F) Relative amount of icellular ATP in 5-8F-vehicle cells, 5-8F-KISS1 cells, 5-8F-1R cells and 5-8F-1R-KISS1 cells. Unpaired t test for (C) and (E) . Data are presented as the mean ± SEM, *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: After blocking with 5% non-fat milk on a shaking bed and then incubated with the primary antibodies against KISS1 (1:1000, rabbit, Thermo Fisher), KISS1R (D9D7C) (1:1000, rabbit, CST), LKB1 (1:1000, rabbit, CST), Phospho-LKB1 (Ser428) (C67A3) (1:1000, rabbit, CST), CaMKK2 (D8D4D) (1:1000, rabbit, CST), AMPKα (D5A2) (1:1000, rabbit, CST), Phospho-AMPKα (Thr172) (D4D6D) (1:1000, rabbit, CST) and GAPDH (1:1000, rabbit, CST) and subsequently incubated with relevant anti-rabbit IgG-HRP (1:5000, CST) secondary antibodies.

Techniques: Over Expression, Phospho-proteomics, Gene Expression, Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot