Wiskott-Aldrich综合征蛋白家族成员3对非小细胞肺癌侵袭转移的影响及机制研究

Effect and mechanism of WASF3-AS1 on invasion and metastasis of non-small cell lung cancer

目的:探究Wiskott-Aldrich综合征蛋白家族成员3(WASF3-AS1)对非小细胞肺癌(NSCLC)侵袭转移的影响及机制。方法:选取26例临床不同病理分期NSCLC患者的癌组织和癌旁组织,以及NSCLC细胞系NCI-H1975细胞和正常肺支气管上皮细胞系HBE细胞为研究对象。实时荧光定量PCR(qRT-PCR)检测上述临床组织和细胞系中WASF3-AS1、赖氨酸特异性组蛋白去甲基化酶1(LSD1)、组蛋白去乙酰化酶6(HDAC6)、E-钙黏蛋白(E-ca)、N-钙黏蛋白(N-ca)的表达。采用CCK-8实验检测细胞增殖。采用划痕实验检测细胞迁移能力。采用Transwell实验检测细胞侵袭能力。采用Western blot检测LSD1、HDAC6、蛋白酪氨酸磷酸酶(PTEN)、蛋白激酶B(AKT)、E-ca和N-ca的表达。结果:与对照组比较,在NSCLC组织和细胞中,WASF3-AS1、LSD1、HDAC6、PTEN和E-ca低表达,AKT和N-ca高表达(均P<0.05);WASF3-AS1过表达能够促进LSD1和HDAC6并抑制PTEN/AKT信号,进而抑制NCI-H1975细胞增殖、迁移、侵袭和上皮间充质转化(均P<0.05)。结论:ASF3-AS1可能通过招募LSD1和HDAC6到靶基因PTEN启动子区发挥抑制NSCLC细胞侵袭转移作用。

Objective:To investigate the effect and mechanism of WASF3-AS1 on the invasion and metastasis of non-small-cell lung cancer(NSCLC).Methods:The cancer tissues and adjacent tissues of 26 NSCLC patients with different clinical pathological stages,as well as NSCLC cell line NCI-H1975 cells and normal lung bronchial epithelial cell line HBE cells were selected as the research objects.Real-time fluorescent quantitative PCR(qRT-PCR)was used to detect the expression of WASF3-AS1,lysine-specific histone demethylase 1(LSD1),histone deacetylase 6(HDAC6),E-cadherin(E-ca)and N-cadherin(N-ca)in the above clinical tissues and cell lines.The CCK-8 experiment was used to detect cell proliferation.Scratch-wound experiment was used to detect cell migration ability.Transwell experiment was used to detect cell invasion ability.Western blot was used to detect the expression of LSD1,HDAC6,protein tyrosine phosphatase(PTEN),protein kinase B(AKT),E-ca and N-ca.Results:Compared with the control group,in NSCLC tissues and cells,WASF3-AS1,LSD1,HDAC6,PTEN and E-ca were lowly expressed,and AKT and N-ca were highly expressed(all P<0.05);Overexpression of WASF3-AS1 could promote LSD1 and HDAC6 and inhibit PTEN/AKT signaling,thereby inhibiting the proliferation,migration,invasion and epithelial-mesenchymal transformation of NCI-H1975 cells(all P<0.05).Conclusion:ASF3-AS1 may inhibit the invasion and metastasis of NSCLC cells by recruiting LSD1 and HDAC6 to the promoter region of the target gene PTEN.