Effects of zinc finger transcription factor 580 in regulating autophagy and extracelluar matrix secretion of PANC1 cells
10.3760/cma.j.cn115667-20250407-00042
- VernacularTitle:锌指转录因子580在胰腺癌细胞PANC1自噬和细胞外基质分泌中的调控作用
- Author:
Baiqing LIU
1
;
Wei XU
1
;
Wencheng ZHANG
1
;
Shihai XIA
1
Author Information
1. 中国人民武装警察部队特色医学中心，天津 300162
- Publication Type:Journal Article
- Keywords:
Pancreatic neoplasms;
Tumor cells, cultured;
Transcription factors;
Autophagy;
Extracellular matrix
- From:
Chinese Journal of Pancreatology
2025;25(5):355-360
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the regulatory effect of zinc finger transcription factor 580 (ZNF580) gene on autophagy and extracellular matrix (ECM) secretion in human pancreatic cancer cells PANC1.Methods:PANC1 cells were transfected with 500 ng/ml short hairpin RNA-ZNF580 (shRNA-ZNF580) and a ZNF580 expression vector with a green fluorescent protein reporter gene (GFP-ZNF580) using lentiviral transfection to establish the ZNF580-silenced group and ZNF580-overexpression group, respectively. PANC1 cells were treated with 10 mmol/L rapamycin (RA), a cell autophagy inducer, and the autophagy inhibitor LY294002 for 2 hours to construct the autophagy-induced group and autophagy-inhibited group, respectively. The autophagy inhibition+ZNF580 silencing group was established by transfecting PANC1 cells with 500 ng/ml sh-ZNF580 using lentiviral transfection while simultaneously adding 10 mmol/L LY294002. PANC1 cells cultured in conventional medium served as control group. The expression levels of ZNF580 protein and autophagy-related proteins ATG7 and LC3 in PANC1 cells from each group were detected by Western blot. The expression changes of ECM secretion-related markers type I collagen (Col-Ⅰ), Col-Ⅲ, fibronectin (FN), and tumor necrosis factor-α (TNF-α) in PANC1 cells were measured by ELISA.Results:Compared with control group, the protein expression levels of ATG7, LC3-Ⅰ, and LC3-Ⅱ in PANC1 cells of the ZNF580-silenced group were significantly decreased (0.40±0.04 vs 0.81±0.13, 0.66±0.08 vs 2.0±0.45, 0.78±0.10 vs 1.89±0.23), while they were significantly increased in the ZNF580-overexpression group (2.07±0.17 vs 0.83±0.09, 1.21±0.37 vs 0.88±0.09, 0.77±0.16 vs 0.37±0.06). The protein expression level of ZNF580 in PANC1 cells of the autophagy inhibition group was significantly down-regulated compared with the control group (0.40±0.15 vs 1.07±0.18), while it was significantly up-regulated in the autophagy induction group (1.59±0.25 vs 0.67±0.09). Compared with the control group, the levels of extracellularly secreted Col-Ⅰ, Col-Ⅲ, FN, and TNF-α in PANC1 cells were significantly decreased in the ZNF580-silenced group (5.02±0.81 vs 8.38±0.83, 6.17±0.83 vs 10.73±1.69, 28.66±2.47 vs 45.20±4.31, 10.09±1.32 vs 19.48±2.77), which were significantly increased in the ZNF580-overexpression group (19.28±2.05 vs 8.38±0.83, 28.29±5.96 vs 10.73±1.69, 103.22±6.37 vs 45.20±4.31, 46.78±6.96 vs 19.48±2.77), significantly decreased in the autophagy inhibition group (5.10±0.66 vs 9.01±1.24, 7.22±0.67 vs 11.83±1.71, 28.45±2.82 vs 43.51±4.38, 12.16±2.13 vs 20.53±3.65, respectively), and significantly increased in the autophagy induction group (20.49±3.68 vs 9.01±1.24, 26.58±3.96 vs 11.83±1.71, 73.18±7.15 vs 43.51±4.38, 41.11±8.87 vs 20.53±3.65). Compared with the autophagy inhibition group and the ZNF580-silenced group, the levels of extracellularly secreted Col-Ⅰ, Col-Ⅲ, FN, and TNF-α in PANC1 cells of the autophagy inhibition+ZNF580 silencing group were significantly decreased (Col-Ⅰ: 3.36±1.25 vs 5.73±0.62 and 5.57±0.35; Col-Ⅲ: 4.15±0.16 vs 6.24±0.90 and 6.71±0.34; FN: 18.31±2.00 vs 26.46±1.18 and 27.09±2.01; TNF-α: 6.81±0.46 vs 9.96±1.87 and 10.62±0.65). All the above differences were statistically significant (all P value <0.05). Conclusions:The transcription factor ZNF580 could positively regulate the levels of autophagy and ECM secretion in PANC1 cells. The combined application of ZNF580 gene silencing and autophagy inhibitors can significantly inhibit ECM secretion in PANC1 cells.
