Research on the function and related mechanism of P27 gene in the intervertebral disc degeneration of mice

Abstract

This study aims to investigate the function and related mechanism of P27 gene in intervertebral disc (IVD) degeneration of mice. X-ray, immunohistochemical staining, and alkaline phosphatase (ALP) histochemical staining were used to analyze the phenotypic difference of the intervertebral discs of 4-week-old mice with P27 gene knockout (P27−/−) and wild-type (WT) mice in the same brood. Protein in the intervertebral disc was extracted and western blot analysis was employed to detect the changes in the expression of related molecules in the Shh-signal pathways, including Shh, Patched, Smoothened and Gli2. As a result, the ALP histochemical staining revealed that the ALP-positive area of mice in the P27−/− group was obviously increased compared to the 4-week-old mice of the same brood in the WT group. In addition, the Col-I immunohistochemical staining showed that the Col-I-positive area of mice in the P27−/− group was significantly increased compared to mice in the WT group. Furthermore, Smo-positive cell rate of mice in the P27−/− group was apparently increased compared to mice in the WT group. Western blot analysis revealed that in terms of changes of protein expression levels of Shh, Patched, Smoothened and Gli2 in the intervertebral disc, protein expression levels of Shh, Patched, Smoothened and Gli2 of mice in the P27−/− group were significantly increased compared to those of mice in the WT group. The results show that P27 deficiency activates the expression of Shh-signal pathway and promotes the proliferation of osteoblast, thus, playing a role in promoting IVD degeneration, which provides a scientific and reliable experimental basis for the treatment of the IVD degeneration-related diseases in clinical practice.

Delta Like-1 Gene Mutation: A Novel Cause of Congenital Vertebral Malformation

“…This effect is modulated by type I collagen, osteoblast number and activity, and mRNA expression of RUNX2, osteocalcin, and BMP2 (Yin et al, 2014). The proliferation of osteoblasts may also be regulated via the Sonic hedgehog signaling pathway, as demonstrated in p27-deficient mice (Liu et al, 2017). This may explain the observed downregulation of p27 in the affected siblings, possibly to counteract the negative effect of p27 and enhance bone formation.…”

Key LncRNAs Associated With Oxidative Stress Were Identified by GEO Database Data and Whole Blood Analysis of Intervertebral Disc Degeneration Patients

“…Overexpression of MIR100HG significantly increased cell proliferation, and mir-5590-3p expressed the opposite effect (Wang et al, 2018b;Chen et al, 2020). The lack of the P27 gene activates the expression of Shh signaling pathway and promotes the proliferation of osteoblasts, thus playing a role in promoting IDD (Liu et al, 2017). MIR100HG may promote the proliferation, migration, and invasion of laryngeal squamous cell carcinoma (LSCC) through down-regulation of mir-204-5p (Huang et al, 2019).…”

Constitutive and conditional gene knockout mice for the study of intervertebral disc degeneration: Current status, decision considerations, and future possibilities

“…When Tnmd gene is mutated in IVDs, Chm1 may play a compensatory role. 42 Many other genes have similar functions 43 , 44 , 45 , 46 , 47 , 48 , 49 , 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 , 60 , 61 , 62 , 63 , 64 , 65 (Table 2 ).…”

“…When Tnmd gene is mutated in IVDs, Chm1 may play a compensatory role 42 . Many other genes have similar functions 43–65 (Table 2).…”