Gong Xin, Liu Jing, Zhang Dan, Yang Dawei, Min Zhihui, Wen Xiaoxing, Wang Guifang, Li Huayin, Song Yuanlin, Bai Chunxue, Li Jing, Zhou Jian
Department of Pulmonary Medicine, Shanghai Respiratory Research Institute, Zhongshan Hospital, Fudan University, Shanghai, China.
Department of Pathology, The Affiliated Yantai Yuhuangding Hospital, Qingdao University, Yantai, China.
PLoS One. 2017 Aug 3;12(8):e0182410. doi: 10.1371/journal.pone.0182410. eCollection 2017.
Chemotherapy drugs, such as cisplatin (DDP), improve the survival of patients with lung cancer by inducing apoptosis in cancer cells, which quickly develop resistance to DDP through uncharacterized mechanisms. Glioma Pathogenesis-Related Protein 1 (GLIPR1) plays an important role in cell proliferation, migration and apoptosis. However, the expression and function of GLIPR1 in mediating DDP resistance in human lung adenocarcinoma A549/DDP and human large cell lung cancer H460/DDP cells has not yet been reported.
In this study, real-time PCR (RT-PCR) and western blot were used to examine the mRNA and protein expression of GLIPR1, respectively. Bright-field microscopy, the cell counting kit-8 (CCK-8) assay, flow cytometry analysis and JC-1 dye were used to measure the cellular morphology, proliferation, apoptosis and mitochondrial membrane potential, respectively.
Compared to human lung adenocarcinoma A549 cells, the mRNA and protein expression of GLIPR1 were significantly increased in DDP-resistant A549/DDP cells (p < 0.05). Similarly, the mRNA level of GLIPR1 in DDP-resistant H460/DDP cells was also significantly higher than that in DDP-sensitive H460 cells (p < 0.05). Silencing of GLIPR1 in A549/DDP and H460/DDP cells led to increased apoptosis via a mitochondrial signaling pathway following incubation with various concentrations of DDP. Furthermore, GLIPR1 downregulation markedly reduced the protein expression of Bcl-2, and increased the cleaved Poly (ADP-Ribose) Polymerase (PARP) and cleaved caspase-3 in DDP-resistant A549/DDP cells.
In this study, we demonstrated for the first time that GLIPR1 could modulate the response of DDP-resistant A549/DDP and H460/DDP cells to cisplatin. Therefore, GLIPR1 deserves further investigation in the context of none-small lung cancer (NSCLC).
化疗药物，如顺铂（DDP），通过诱导癌细胞凋亡来提高肺癌患者的生存率，但癌细胞会通过未知机制迅速对DDP产生耐药性。胶质瘤发病机制相关蛋白1（GLIPR1）在细胞增殖、迁移和凋亡中起重要作用。然而，GLIPR1在介导人肺腺癌A549/DDP和人肺大细胞癌H460/DDP细胞对DDP耐药中的表达及功能尚未见报道。
本研究分别采用实时荧光定量PCR（RT-PCR）和蛋白质印迹法检测GLIPR1的mRNA和蛋白表达。分别用明场显微镜、细胞计数试剂盒-8（CCK-8）法、流式细胞术分析和JC-1染料检测细胞形态、增殖、凋亡及线粒体膜电位。
与人类肺腺癌A549细胞相比，耐DDP的A549/DDP细胞中GLIPR1的mRNA和蛋白表达显著增加（p<0.05）。同样，耐DDP的H460/DDP细胞中GLIPR1的mRNA水平也显著高于对DDP敏感的H460细胞（p<0.05）。在A549/DDP和H460/DDP细胞中沉默GLIPR1后，用不同浓度的DDP孵育，通过线粒体信号通路导致细胞凋亡增加。此外，GLIPR1下调显著降低了耐DDP的A549/DDP细胞中Bcl-2的蛋白表达，并增加了裂解的聚（ADP-核糖）聚合酶（PARP）和裂解的半胱天冬酶-3。
在本研究中，我们首次证明GLIPR1可以调节耐DDP的A549/DDP和H460/DDP细胞对顺铂的反应。因此，在非小细胞肺癌（NSCLC）的背景下，GLIPR1值得进一步研究。
