TP73-AS1 regulates cell autophagy via PTEN/AKT signaling pathway and participates in the pathogenesis of Parkinson disease
10.3760/cma.j.cn371468-20241129-00571
- VernacularTitle:TP73-AS1调控PTEN/AKT介导的自噬在帕金森病发病机制中的作用
- Author:
Xue ZHANG
1
;
Anmu XIE
Author Information
1. 青岛大学附属医院神经内科，青岛 266003
- Publication Type:Journal Article
- Keywords:
Parkinson disease;
Long non-coding RNA;
p73 antisense RNA 1T;
Phosphatase and tensin homolog/protein kinase B signaling pathway;
Autophagy
- From:
Chinese Journal of Behavioral Medicine and Brain Science
2025;34(4):296-302
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effects of long non-coding RNA(lncRNA) p73 antisense RNA 1T(TP73-AS1) on cell viability, autophagy and phosphatase and tensin homolog/protein kinase B(PTEN/AKT) signaling pathway in Parkinson disease cell model.Methods:SH-SY5Y cells were treated with 1-methyl-4-phenylpyridine(MPP+ ) to establish Parkinson disease cell model.The cultured SH-SY5Y cells were divided into si-NC group (transfected with empty virus negative control), si-NC+ MPP+ group (transfected with empty virus negative control, followed by 1 mol/L MPP+ ), si-TP73-AS1 group (transfected with si-TP73-AS1 target virus), and si-TP73-AS1+ MPP+ group (transfected with si-TP73-AS1 target virus, followed by 1 mol/L MPP+ ). The proliferation rate of SH-SY5Y cells was detected by CCK-8. Western blot method was used to detect the protein expression PTEN, AKT, Beclin1, and p62. qRT-PCR was used to explore the mRNA expression levels of TP73-AS1, PTEN and AKT. Graphpad Prism 5.0 software was used for data analysis. The comparison among multiple groups was conducted by one-way ANOVA, and further pairwise comparisons were conducted by LSD- t test. Results:(1) The results of CCK-8 showed that the cell proliferation rate of SH-SY5Y cells was significantly different among the four groups ( F=425.1, P<0.001).Compared with the si-NC+ MPP+ group ((57.17±3.60)%), the cell proliferation rate of si-TP73-AS1+ MPP+ group was significantly higher((89.33±2.58)%)( P<0.001). (2) qRT-PCR results showed that there were statistical differences in the expression of PTEN mRNA among the four groups ( F=113.4, P<0.001). The mRNA level of PTEN in si-NC+ MPP+ group was higher than that in si-NC group (2.65±0.23, 1.00±0.18, P<0.05). The mRNA level of PTEN in si-TP73-AS1+ MPP+ group was higher than that in si-TP73-AS1 group (1.69±0.14, 1.00±0.15, P<0.05).There was no significant difference in mRNA level of AKT among the 4 groups ( F=58.1, P>0.05). (3) Western blot results showed that there were significant differences in the expressions of PTEN protein and p-AKT/AKT ratio among the 4 groups ( F=176.3, 178.5, both P<0.001). The level of PTEN protein in si-TP73-AS1+ MPP+ group was lower than that in si-NC+ MPP+ group(0.76±0.03, 1.05±0.04, P<0.001).The p-AKT/AKT ratio in si-TP73-AS1+ MPP+ group was higher than that in si-NC+ MPP+ group (0.66±0.04, 0.38±0.02, P<0.001).There were significant differences in the expression of Beclin1 and p62 proteins among the 4 groups ( F=156.2, 182.2, both P<0.001). The expression level of p62 protein in si-TP73-AS1+ MPP+ group was lower than that in si-NC+ MPP+ group (0.72±0.03, 1.06±0.10, P<0.001). The expression level of Beclin1 protein level in si-TP73-AS1+ MPP+ group was higher than that in si-NC+ MPP+ group (0.97±0.06, 0.70±0.06, P<0.001). Conclusion:Silencing of TP73-AS1 can increase the expression of Beclin1 protein and decrease the expression of p62 protein. The mechanism may be related to the down-regulation of PTEN/AKT autophagy pathway.
