长链非编码RNA PAPPA-AS1在人肺腺癌细胞铂类耐药中的作用

目的: 研究长链非编码RNA（long noncoding RNA，lncRNA）PAPPA-AS1在人肺腺癌细胞铂类耐药中的作用，并初步探讨其可能的调控机制。方法: 采用RNA测序（RNA-seq）技术检测人肺腺癌细胞A549及其铂类耐药细胞中lncRNA和mRNA的差异表达谱；qRT-PCR检测lncRNA PAPPA-AS1在A549/DDP和A549/NDP细胞中的表达水平；利用siRNA技术下调铂类耐药人肺癌细胞中PAPPA-AS1的表达，CCK-8实验检测肺腺癌细胞对铂类药物的敏感性；克隆形成实验检测细胞的增殖能力；流式细胞术检测细胞凋亡分布；蛋白质印迹法检测Cleaved Caspase-3、Cleaved Caspase-9、p-PI3K、p-Akt、p-mTOR蛋白在A549/DDP和A549/NDP细胞中的表达水平。 结果： 与亲代敏感细胞A549相比，lncRNA PAPPA-AS1在A549/DDP和A549/NDP细胞中呈高表达；与对照组比较，si-PAPPA-AS1组的肺腺癌细胞对铂类药物的敏感性增强，细胞增殖活力下降，细胞凋亡率增加（P均<0.05）；与对照组比较，si-PAPPA-AS1组的细胞中Cleaved Caspase-3和Cleaved Caspase-9蛋白表达明显升高，磷酸化PI3K、Akt和mTOR蛋白表达明显降低。结论： 下调PAPPA-AS1可以通过抑制PI3K/Akt信号通路逆转肺腺癌细胞对铂类药物的抵抗性。

The role of long noncoding RNA PAPPA-AS1 in platinum-resistant lung adenocarcinoma cells

Objective: This study aims to investigate the role of lncRNA PAPPA-AS1 in platinum-resistant cells of lung adenocarcinoma(LAD) and to explore its possible regulatory mechanism. Methods: The differential expression profiling of lncRNA and mRNA in human lung adenocarcinoma(LAD) parental A549 cells and platinum-resistant cells were detected by RNA-seq; the expression of lncRNA PAPPA-AS1 in A549/DDP and A549/NDP cells were detected by fluorescence quantitative PCR; After using siRNA to down-regulate the expression of PAPPA-AS1 in platinum-resistant lung adenocarcinoma cells, the Chemosensitivity of LAD cells to platinumdrugs was detected by CCK-8 assay;the proliferation ability of platinum-resistant cells was detected by cloning formation assay; flow cytometry was used to detect the apoptosis of platinum-resistant cells; Western blotting was used to detect the expression of Cleaved Caspase-3, Cleaved Caspase-9, p-PI3K, p-Akt and p-mTOR protein levels in A549/DDP and A549/NDP cells. Results: Compared with human LAD parental A549 cells, lncRNA PAPPA-AS1 was highly expressed in A549/DDP and A549/NDP. Compared with the control group, LAD platinum-resistant cells in the si-PAPPA-AS1 group showed increased sensitivity to drugs, decreased cell proliferation, and increased apoptosis (all P<0.05). Compared with the control group, the expression of Cleaved Caspase-3 and Cleaved Caspase-9 protein levels in si-PAPPA-AS1cells were significantly up-regulated, and the expression of phosphorylated PI3K, Akt and mTOR protein levels weredown-regulated. Conclusion: Down-regulation of PAPPA-AS1 can reverse platinumresistance of LAD via inhibiting the PI3K/Akt signaling pathway. ［Key words］lncRNA PAPPA-AS1; platinum resistance; lung cancer; RNA-Seq；cell proliferation; cell apoptosis；PI3K/Akt signaling pathway