Department of General Thoracic Surgery, Faculty of Medicine, Kagawa University, Kagawa, Japan.
Department of General Thoracic Surgery, Faculty of Medicine, Kagawa University, Kagawa, Japan
Anticancer Res. 2020 Feb;40(2):733-741. doi: 10.21873/anticanres.14004.
BACKGROUND/AIM: GPR87 is a member of the cell surface molecular G protein-coupled receptors (GPCR) family and suggested to contribute to the viability of human tumor cells. Its tumor-specific expression and cell surface location make it a potential molecule for targeted therapy. In the present study, we aimed to examine the effect of silencing GPR87 expression and explore the possibility of establishing gene therapy against GPR87-overexpressing lung cancer.
Twenty malignant cell lines were investigated and GPR87-overexpressing H358 and PC9 lung cancer cells were subjected to inhibiting experiments. A short hairpin siRNA targeting the GPR87 gene was transformed into an adenoviral vector (Ad-shGPR87). Real-time RT-PCR and western blot analyses were performed to evaluate gene and protein expression. Tumors derived from human H358 cells were subcutaneously implanted in nude mice for in vivo experiments.
About 50% (10/20) malignant cells showed GPR87-overexpression, especially for lung cancer cells (70%, 7/10). Ad-shGPR87 effectively down-regulated the GPR87 expression, and significantly inhibited the cell proliferation in GPR87-overexpressing H358 and PC9 cells. Treatment with Ad-shGPR87 exerted a significant antitumor effect against the GPR87-expressing H358 xenografts. In addition, the gene expression of H3.3, a recently proved activator for GPR87 transcription, was positively correlated with GPR87 gene expression. Furthermore, a significant decrease of KRAS and c-Myc expression was observed in both cell lines after Ad-shGPR87 infection. In conclusion, GPR87 may play a critical role in cancer cell proliferation, and indicate its potential as a novel target for lung cancer treatment.
背景/目的：GPR87 是细胞表面分子 G 蛋白偶联受体（GPCR）家族的成员，被认为有助于人类肿瘤细胞的存活。其肿瘤特异性表达和细胞表面位置使其成为靶向治疗的潜在分子。在本研究中，我们旨在研究沉默 GPR87 表达的效果，并探索针对 GPR87 过表达肺癌建立基因治疗的可能性。
研究了 20 种恶性细胞系，并对 GPR87 过表达的 H358 和 PC9 肺癌细胞进行了抑制实验。针对 GPR87 基因的短发夹 siRNA 被转化为腺病毒载体（Ad-shGPR87）。实时 RT-PCR 和 Western blot 分析用于评估基因和蛋白表达。源自人 H358 细胞的肿瘤被皮下植入裸鼠进行体内实验。
约 50%（10/20）的恶性细胞表现出 GPR87 过表达，特别是肺癌细胞（70%，7/10）。Ad-shGPR87 有效地下调了 GPR87 的表达，并显著抑制了 GPR87 过表达的 H358 和 PC9 细胞的增殖。Ad-shGPR87 对 GPR87 表达的 H358 异种移植物具有显著的抗肿瘤作用。此外，最近被证明是 GPR87 转录激活剂的 H3.3 基因的表达与 GPR87 基因的表达呈正相关。此外，在感染 Ad-shGPR87 后，两种细胞系中的 KRAS 和 c-Myc 表达均显著下降。总之，GPR87 可能在癌细胞增殖中发挥关键作用，并表明其作为肺癌治疗新靶点的潜力。
