SPARC knockout affects the development of metabolic fatty liver related liver cancer by regulating liver lipid metabolism

Abstract

Abstract: Objective To investigate the impact of secreted protein acidic and rich in cysteine (SPARC) gene knockout on the development of metabolic fatty liver related liver cancer by regulating liver lipid metabolism.Methods The study samples were selected from 17 adult healthy rats as the control group and 17 SPARC knockout rats as the study group. Hepatocytes were extracted from one rat each from both groups for in vitro experiments. The remaining animals were treated with streptozotocin Injection and high-fat diet to induce metabolic fatty liver related liver cancer. The pathological changes, the lipid deposition, lipid synthesis, lipid metabolism, cell viability related indicators and the protein expression of AMPK and α-Tubulin in the livers of two groups of rats were studied.Results At the 16th week, all rats from both groups developed liver cancer. The average diameter of liver cancers in the study group was (1.39±0.14) mm, which was significantly larger than that of (1.01±0.09) mm in the control group; the NAS score showed that the fatty degeneration score of the study group was (2.71±0.25), which was significantly higher than that of (1.55±0.19) of the control group. The score of lobular inflammation in the study group was (0.74±0.09), which was significantly lower than that of (1.27±0.11) in the control group. The results of H&E staining showed that the portal vein inflammation of the rats in the study group was weaker; the results of Sirius red staining showed that the liver fibrosis of the rats in the study group was more severe than that of the control group; in the in vitro study, the results of oil red O staining showed that the lipid deposits in the cells of the study group increased, and the expression of GL-TAG (23.15±5.39 vs. 5.73±1.47) and GPL (0.29±0.03 vs. 0.23±0.02) were significantly higher than those of the control group; the expression of [U-14C]-glycerol in the liver cells of the study group was significantly higher than that of the control group; The relative expressions of PPARγ (1.95±0.20 vs. 1.01±0.11), PPARα(1.27±0.12 vs. 0.98±0.09), CD36 (2.24±0.26 vs. 0.98±0.09), FABP4 (1.92±0.19 vs. 0.99±0.10), FABP5 (2.31±0.27 vs. 0.97±0.10), ACC1 (2.52±0.27 vs. 1.01±0.11), FASN (1.28±0.13 vs. 0.99±0.09), SREBP1 (3.95±0.43 vs. 1.02±0.11), LXRα (1.52±0.16 vs. 0.97±0.09), CPT1 (2.76±0.29 vs. 1.02±0.11), LIPIN (2.49±0.26 vs. 1.01±0.11), ACOX1 (2.69±0.28 vs. 1.01±0.11) mRNAs in rat livers of the study group were significantly lower than that of the control group (all P<0.05). There was no statistically significant difference in ACADSB mRNA expression in rat livers from these two groups (P>0.05); the results of acridine orange-ethidium bromide staining showed that cells from the study group had higher viability than those from the control group, and had a significantly higher cell survival rate than those from the control group {(97.89±1.44)% vs. (48.47±1.21)%}; Immunofluorescence detection results showed that cells in the study group had increased nuclear localization of SREBP in hepatocytes. By Western blot analysis it was shown that the expression of AMPK and α-Tubulin protein in the study group was lower than that of the control group.Conclusion SPARC knockout can affect the development of metabolic fatty liver-related liver cancer by regulating lipid metabolism, cell viability and lipid accumulation in the liver cells in rats.

Key words: secreted protein acidic and rich in cysteine, liver, lipid metabolism, metabolic fatty liver, liver cancer

CHANG Ye-fei, ZHENG Sheng, HU Tao, HONG Ying, ZHONG Yong, ZHANG Xiao-jun. SPARC knockout affects the development of metabolic fatty liver related liver cancer by regulating liver lipid metabolism[J]. Chinese Hepatolgy, 2022, 27(12): 1309-1313.

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