靶向沉默KIAA1199基因表达对人非小细胞肺癌A549细胞生物学行为的影响

EFFECTS OF SILENCING EXPRESSION OF KIAA1199 GENE ON BIOLOGICAL BEHAVIORS OF HUMAN NON-SMALL CELL LUNG CANCER A549 CELLS

目的探究靶向沉默KIAA1199基因表达对人非小细胞肺癌A549细胞生物学行为的影响。方法采用实时荧光定量PCR(qRT-PCR)和蛋白印迹(western blot)检测肺正常上皮细胞系BEAS-2B和肺癌细胞系(A549、H332和H1299)中KIAA1199 mRNA和蛋白的表达;以小干扰RNA(siRNA)技术沉默A549细胞中KIAA1199的表达,并经qRT-PCR和Weatern blot验证KIAA1199表达变化;CCK-8实验、流式细胞仪、Transwell小室和划痕实验检测细胞增殖、凋亡、侵袭和迁移能力,Weatern blot检测细胞中相关蛋白的表达。结果与正常上皮细胞系BEAS-2B比较,肺癌细胞系A549、H332和H1299细胞中KIAA1199 mRNA和蛋白的表达均升高(P<0.05);与空白组比较,KIAA1199-siRNA组KIAA1199mRNA和蛋白表达、细胞A值、穿膜细胞数和迁移率、细胞中PCNA、Cyclin D1、p-p38/p38、p-JNK/JNK及p-ERK1/2/ERK1/2表达均降低,细胞凋亡率和细胞中caspase-3和caspase-9蛋白表达增加(均P<0.05),但空白组和sc-siRNA组上述指标的比较,差异无统计学意义(P>0.05)。结论沉默KIAA1199基因表达可抑制A549细胞的增殖、侵袭与迁移,诱导其凋亡,可能与阻滞MAPK信号通路有关。

Objective Objective To explore the effects of silencing the expression of KIAA1199 gene on biological behaviors of human non-small cell lung cancer(NSCLC) A549 cells.Methods The expression of KIAA1199 mRNA and protein in normal lung epithelial cell line BEAS-2 B and lung cancer cell lines(A549,H332,H1299) was detected by real-time quantitative PCR(qRT-PCR)and western blot.The expression of KIAA1199 in A549 cells was silenced by small interfering RNA(siRNA) technology.The changes in expression of KIAA1199 were verified by qRT-PCR and western blot.The cells proliferation, apoptosis, invasion andmigration abilities were detected by CCK-8,flow cytometry, Transwell chamber assay and scratch test.The expression of related proteins in cells was detected by western blot.Results Compared with normal epithelial cell line BEAS-2 B,the expression of KIAA1199 mRNA and protein in lung cancer cell lines A549,H332 and H1299 was all increased(P<0.05).Compared with blank group, expression of KIAA1199 mRNA and protein, A value of cells, number of transmembrane cells, migration rate, expression of PCNA,Cyclin D1,p-p38/p38,p-JNK/JNK and p-ERK1/2/ERK1/2 in cells were all decreased, while apoptosis rate, expression of caspase-3 and caspase-9 proteins in cells were increased in KIAA1199-siRNA group(P<0.05).However, there were no significant differences in the above indexes between blank group and sc-siRNA group(P>0.05).Conclusion Silencing the expression of KIAA1199 gene can inhibit proliferation, invasion and migration of A549 cells, and induce their apoptosis, which may be related to blocking MAPK signal pathway.