Journal: Journal of Translational Medicine

Article Title: Adiponectin suppresses tumor growth of nasopharyngeal carcinoma through activating AMPK signaling pathway

doi: 10.1186/s12967-022-03283-0

Figure Lengend Snippet: Circulating adiponectin levels and risk of nasopharyngeal carcinoma in retrospective and prospective cohorts

Article Snippet: Recombination human full-length adiponectin was dissolved in deionized water to prepare a working stock solution of approximately 0.5 mg/mL (BioVendor, Brno, Czech Republic).

Techniques:

Journal: Journal of Translational Medicine

Article Title: Adiponectin suppresses tumor growth of nasopharyngeal carcinoma through activating AMPK signaling pathway

doi: 10.1186/s12967-022-03283-0

Figure Lengend Snippet: Adiponectin suppresses nasopharyngeal carcinoma growth. A , B 1 × 10 6 CNE-2 cells were injected subcutaneously into 5- to 6- week-old adiponectin-deficient nude mice, or the control nude mice ( n = 6 per group). Tumor growth were monitored by measuring the tumor volume for 10 days. Next, the mice were sacrificed, and tumors were collected, measured, weighed. C The plate colony assay was performed to determine colony-formation ability. CNE-2 and C666-1 cells were treated with various concentrations of adiponectin for 7 days. Graphs show the number of colonies. D EdU incorporation assay was performed to determine cell proliferation. CNE-2 and C666-1 cells were treated with various concentrations of adiponectin for 48 h. Bars: 50 μm. Graphs show the relative cell proliferation percentage. E , F CNE-2 and C666-1 cells were incubated with adiponectin (40 μg/mL) for 48 h. At the end of incubation, the cells were collected for FACS analysis. G Western blot analysis of p-AMPKα (T172), p-LKB1 and p-ERk1/2 in cultured CNE-2 and C666-1 cells after the treatment with adiponectin (40 μg/mL) for 30 min. Results are presented as mean ± SD of three independent experiments performed in triplicate. * P < 0.05, ** P < 0.01, *** P < 0.001, # P < 0.0001

Article Snippet: Recombination human full-length adiponectin was dissolved in deionized water to prepare a working stock solution of approximately 0.5 mg/mL (BioVendor, Brno, Czech Republic).

Techniques: Injection, Colony Assay, Incubation, Western Blot, Cell Culture

Journal: Journal of Translational Medicine

Article Title: Adiponectin suppresses tumor growth of nasopharyngeal carcinoma through activating AMPK signaling pathway

doi: 10.1186/s12967-022-03283-0

Figure Lengend Snippet: Adiponectin suppresses proliferation of NPC cells via AMPK activation. A , B CNE-2 and C666-1 cells were pretreated with compound C (10 mM) followed by treatment with adiponectin (40 μg/mL) for 24 h. Cell cycle was then analyzed using flow cytometer. # P < 0.05 and ## P < 0.01 compared to cells treated with adiponectin but not ComC; ** P < 0.01 compared with cells treated without adiponectin and ComC. C CNE-2 and C666-1 cells viability was determined after treatment with or without adiponectin for 48 h in the presence or absence of ComC (10 μM). ** P < 0.01, *** P < 0.001. D CNE-2 and C666-1 cells were pretreated with compound C (10 mM), p-AMPKα (Thr172) protein level was then determined by Western blot analysis after the treatment with adiponectin (40 μg/mL) for 30 min; cyclin D1, p21, and p27 protein level was then determined by Western blot analysis after the cells were exposed to adiponectin (40 μg/mL) for 48 h. E Quantitative analysis of p-AMPKα level was performed by densitometric analysis. Results are presented as mean ± SD of three independent experiments performed in triplicate. ** P < 0.01, *** P < 0.001

Article Snippet: Recombination human full-length adiponectin was dissolved in deionized water to prepare a working stock solution of approximately 0.5 mg/mL (BioVendor, Brno, Czech Republic).

Techniques: Activation Assay, Flow Cytometry, Western Blot

Journal: Journal of Translational Medicine

Article Title: Adiponectin suppresses tumor growth of nasopharyngeal carcinoma through activating AMPK signaling pathway

doi: 10.1186/s12967-022-03283-0

Figure Lengend Snippet: AdipoR1 and AdipoR2 mediate the anti-proliferative effect of adiponectin in NPC cells. A , B Expression of AdipoR1 and AdipoR2 was determined by qRT-PCR and Western blot in NPC cell lines. C , D CNE-2 and C666-1 cells were transfected with 50 μM siRNAs of NC, AdipoR1, or AdipoR2. The relative amounts of each AdipoR1/R2 mRNA against β-actin were measured with qRT-PCR. E Effect of the knockdown of AdipoR1 and AdipoR2 expression on the cell viability of CNE-2 and C666-1 cells treated with or without 40 μg/mL adiponectin for 48 h. F CNE-2 and C666-1 cells were transfected with AdipoR1, AdipoR2 siRNA or NC siRNA and treated with 40 μg/mL adiponectin for 30 min. AMPKα and p-AMPKα (T172) protein levels were determined by Western blot analysis. Quantitative analysis of p-AMPKα level was performed by densitometric analysis and shown in the below part. Results are presented as mean ± SD of three independent experiments performed in triplicate. ** P < 0.01, *** P < 0.001

Article Snippet: Recombination human full-length adiponectin was dissolved in deionized water to prepare a working stock solution of approximately 0.5 mg/mL (BioVendor, Brno, Czech Republic).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Transfection