Effect of protein tyrosine phosphatase receptor R-type on malignant biological behavior of glioma cells

doi:10.11958/20240831

Abstract

Abstract:

Objective To explore effects of protein tyrosine phosphatase receptor type R (PTPRR) on malignant biological behavior of glioma cells.Methods UALCAN website was used to analyze the expression of PTPRR in glioma samples from the Cancer Genome Atlas (TCGA) database. The fresh brain glioma tissue samples of 20 patients with glioma and normal brain tissue samples of 20 patients with traumatic cerebral hemorrhage removed during operation were obtained. Human glioma cell line and human astrocyte cell line were cultured. U87 cells were respectively transfected with PTPRR over-expression plasmid pcDNA3.1/PTPRR (pcDNA3.1/PTPRR group) and blank control plasmid pcDNA3.1 (pcDNA3.1 group). The expression of PTPRR mRNA in different glioma cell lines and glioma tissue samples were detected by quantitative real-time PCR. Cell activity was detected by CCK-8 assay. Cell proliferation capacity was detected by colony formation assay. Cell migration and invasion ability were detected by transwell assay. Meanwhile, the cell apoptosis experiment was performed.Results The result of UALCAN database analysis indicated that the expression of PTPRR in glioma was lower than that in normal brain tissue, and the expression of PTPRR was lower in high grade gliomas than that of low grade gliomas. The survival time of glioma patients with low/medium PTPRR expression was shorter than that of patients with high PTPRR expression. The result quantitative real-time PCR showed that the expression levels of PTPRR mRNA in different glioma cell lines and glioma tissue were decreased. CCK-8 assay showed that the cell viability at 24 h and 48 h after over-expression of PTPRR was decreased respectively compared with the pcDNA3.1 group. Results of colony formation assay, Transwell migration and invasion assays showed that the number of colony formation, migration and invasion ability of glioma cells after over-expression of PTPRR were lower than the pcDNA3.1 group. The result of apoptosis experiment showed that the apoptosis rate of glioma cells after over-expression of PTPRR was increased compared with that of the pcDNA3.1 group.Conclusion The expression of PTPRR in glioma is lower. PTPRR acted as a tumor suppressor gene inhibits the activity, proliferation, migration and invasion of glioma cells, and promotes glioma cell apoptosis.

Key words: glioma, cell proliferation, cell movement, neoplasm invasiveness, protein tyrosine phosphatase receptor type R

CLC Number:

R739.41

Figures/Tables 7

Tab.1 Primer sequence

基因名称	引物序列（5′→3′）	产物大小/bp
PTPRR	上游：ACTGTCCGAGGCAAAGAC 下游：CAGAGGTAGCGGTGGTAG	125
β-actin	上游：CGTGACATTAAGGAGAAGCTG 下游：CTAGAAGCATTTGCGGTGGAC	501

Fig.1 UALCAN website used to analyze the expression of PTPRR in glioma

Tab.2 Comparison of PTPRR mRNA expression, cell activity and cell proliferation ability between the two groups （n=3，$\bar{x}$±s）

组别	PTPRR mRNA	细胞活性/OD值			细胞集落数/个
组别	PTPRR mRNA	0 h	24 h	48 h	细胞集落数/个
pcDNA3.1组	1.00±0.23	1.00±0.02	1.00±0.02	1.00±0.01	91.33±6.55
pcDNA3.1/PTPRR组	34.77±3.22	0.97±0.02	0.81±0.02	0.74±0.06	24.00±2.94
t	14.900^＊	0.672	11.520^＊	12.660^＊	12.340^＊

Fig.2 Effects of PTPRR on the cell proliferation of glioma cells investigated by colony formation assays

Tab.3 Comparison of cell migration and invasion after over-expression of PTPRR （n=3，$\bar{x}$±s）

组别	迁移数/（个/视野）	侵袭数/（个/视野）
pcDNA3.1组	116.00±3.74	77.67±4.99
pcDNA3.1/PTPRR组	78.00±3.27	41.33±2.87
t	7.703^＊	12.760^＊

Fig.3 Effects of PTPRR on the migration and invasion of glioma cells investigated by Transwell assays

Fig.4 The effect of PTPRR on apoptosis of glioma cells detected by apoptosis experiment

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