Experimental study on UBQLN2 induced-radioresistance in esophageal squamous cell carcinoma cells by upregulating purine metabolism levels
10.3760/cma.j.cn112271-20240220-00065
- VernacularTitle:UBQLN2通过上调嘌呤代谢水平诱导食管鳞癌细胞放射抵抗的实验研究
- Author:
Yanli MA
1
;
Jialin WANG
;
Rong MA
;
Wenyan PAN
;
Zhoulan BAI
;
Yanyang WANG
Author Information
1. 宁夏医科大学总医院肿瘤医院放疗科，银川 750001
- Keywords:
Esophageal squamous cell carcinoma (ESCC);
UBQLN2;
Radiosensitivity;
Purine metabolism
- From:
Chinese Journal of Radiological Medicine and Protection
2024;44(11):909-916
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To observe the effect of ubiquilin 2 gene (UBQLN2) on the radiosensitivity of human esophageal squamous cell carcinoma cells (ESCC) of EC109 and KYSE30, and explore underlying molecular mechanism.Methods:siRNA and lentivirus transfection techniques were used to establish UBQLN2-knockdown and UBQLN2-overexpression cell lines. The cells were irradiated with a dose of 4 Gy X-rays. UPLC-Q-TOF-MS technique was used for metabolite difference analysis and KEGG pathway analysis. The results of metabonomics analyses were verified by qRT-PCR assay. The influence of UBQLN2 level on the radiosensitivity of ESCC was confirmed by CCK-8 cell proliferation assay and clone formation assay and further verified by treating the cells with metabolic enzyme inhibitors and exogenous metabolites.Results:Compared with irradiation alone, down-regulating UBQLN2 in EC109 and KYSE30 cell lines reduced the cell survival by 32.29% and 16.42% ( t=5.35, 4.88, P<0.05), and reduced the clone formation rate by 11.07% and 7.47% after 4 Gy irradiation, respectively ( t=4.18, 5.09, P<0.05). On the contrary, up-regulating UBQLN2 in EC109 and KYSE30 cell lines increased the survival rate by 14.07% and 10.64% ( t=5.88, 4.21, P<0.05), and increased the clone formation rate by 6.53% and 7.87% after 4 Gy irradiation, respectively ( t=8.60, 8.26, P<0.05). Metabonomics study showed that the purine metabolic pathway was significantly enriched after down-regulating UBQLN2 in EC109 cell. The qRT-PCR experiment showed a positive correlation between the expression level of UBQLN2 and the mRNA level of five purine metabolism enzymes. The viability of irradiated UBQLN2-overexpression cells decreased by 18.28% and 25.58%, respectively ( t=7.76, 10.95, P<0.05), and the clone formation rate decreased by 9.33% and 9.93%, respectively ( t=5.97, 8.02, P<0.05) after adding mycophenolic acid(MPA). However, the survival rate of cells increased by by 8.28% and 10.74% ( t=2.83, 6.20, P<0.05), and the clone formation rate increased by 7.33% and 5.80%, respectively ( t=7.16, 5.49, P<0.05), when exogenous supplementation of nucleotides (ATP + GTP) were added. Conclusion:The expression level of UBQLN2 was negatively related to the radiosensitivity of ESCC by up-regulating purine metabolism.
