Effect of protein kinase N1 on the biological behavior of human glioma

Yu⁃bing HAO, An⁃ling ZHANG, Hu WANG, Guang⁃xiu WANG, Zhao GAO, Zhi⁃fan JIA

Abstract


Objective To study the expression of protein kinase N1 (PKN1) in glioma and its influence on the biological behavior of glioma cells. Methods The expression of PKN1 gene was verified via GEPIA database, and LN229, U87 and A172 gliomas cells were detected by Western blotting. A172 cells were transfected with negative control sequence (siRNA⁃NC group) and PKN1 small interference RNA (siRNA) sequence (siRNA⁃PKN1 group). CCK⁃8 assay, flow cytometry, scratch assay and Transwell assay were applied to detect proliferation, apoptosis rate, migration ability and invasion ability, respectively. Results Analysis of GEPIA database showed that the expression of PKN1 mRNA in glioblastoma tissues (163 cases) was significantly higher than that in nontumorous brain tissues (207 cases, P < 0.05). Western blotting showed that the relative expression of PKN1 protein in A172 cells was higher than that in U87 cells (t = 3.096, P = 0.036) and LN229 cells (t = 8.994, P = 0.000). The proliferation ability of A172 cells in siRNA⁃PKN1 group was lower than that in siRNA⁃NC group at day 3-6 after PKN1 gene was knocked down (t = 3.275, P = 0.031; t = 5.949, P = 0.004; t = 10.430, P = 0.000; t = 6.645, P = 0.003). The scratch healing rate of A172 cells in both groups increased in different degree over time (F = 249.993, P = 0.000). After PKN1 gene was knocked down, both the scratch healing rate (F = 97.811, P = 0.000) and the invasion cells number (t = 5.840, P = 0.004) of A172 cells were lower than that of siRNA ⁃ NC group, while the apoptosis rate of A172 cells was higher than that of siRNA⁃NC group (t = 5.461, P = 0.006). Conclusions The expression of PKN1 mRNA in glioma was higher than that in nontumorous brain tissue, and knockout of PKN1 gene could inhibit the proliferation, migration and invasion of tumor cells, and induce the apoptosis of tumor cells.

DOI:10.3969/j.issn.1672⁃6731.2020.12.009

Keywords


Glioma; Protein kinases; Cell proliferation; Apoptosis; Cell movement; Neoplasm invasiveness; Immunoblotting; Flow cytometry

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