摘要：

目的 探讨蛋白激酶N1（PKN1）在胶质瘤组织中的表达变化及其对胶质瘤细胞生物学行为的影响。方法 检索GEPIA数据库验证PKN1基因表达情况，Western blotting法检测胶质瘤细胞系LN229、U87和A172的PKN1蛋白表达量。采用无意义序列（siRNA-NC组）和PKN1小干扰RNA序列（siRNA-PKN1组）转染A172细胞，并通过CCK-8实验、流式细胞术、细胞划痕实验和Transwell实验检测肿瘤细胞增殖能力、凋亡率、迁移能力和侵袭能力。结果 经检索GEPIA数据库发现胶质母细胞瘤组织（163例）PKN1 mRNA表达量高于正常脑组织（207例，P<0.05）。Western blotting检测显示，A172细胞PKN1蛋白相对表达量高于U87细胞（t=3.096，P=0.036）和LN229细胞（t=8.994，P=0.000）。敲低PKN1基因表达第3~6天siRNA-PKN1组A172细胞增殖能力低于siRNA-NC组（t=3.275，P=0.031；t=5.949，P=0.004；t=10.430，P=0.000；t=6.645，P=0.003）；两组A172细胞划痕愈合率随时间推移具有不同程度增加（F=249.993，P=0.000），敲低PKN1基因表达后，A172细胞划痕愈合率（F=97.811，P=0.000）和细胞穿膜数目（t=5.840，P=0.004）均低于siRNA-NC组，而A172细胞凋亡率高于siRNA-NC组（t=5.461，P=0.006）。结论 胶质瘤PKN1 mRNA表达量高于正常脑组织，敲低PKN1基因表达可抑制肿瘤细胞增殖、迁移和侵袭，同时诱导肿瘤细胞凋亡。

关键词: 神经胶质瘤, 蛋白激酶类, 细胞增殖, 细胞凋亡, 细胞运动, 肿瘤侵润, 免疫印迹法, 流式细胞术

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.

Key words: Glioma, Protein kinases, Cell proliferation, Apoptosis, Cell movement, Neoplasm invasiveness, Immunoblotting, Flow cytometry