Li Tian-Jiao, Huang Yan-Hong, Chen Xi, Zhou Zhou, Luo Si-Wei, Feng Dan-Dan, Han Jian-Zhong, Luo Zi-Qiang
Grade 2011 of Clinical Medicine, Xiangya School of Medicine, Central South University, Changsha 410013, China.
Department of Physiology, Xiangya School of Medicine, Central South University, Changsha 410013, China.
Sheng Li Xue Bao. 2015 Oct 25;67(5):513-20.
This study aims to detect the expression of metabotropic glutamate receptors (mGluRs) in lung carcinoma A549 cells, and to investigate the effects of mGluR8 and mGluR4 activation on the growth of A549 cells in vitro. The mRNA expression levels of the 8 subtypes of mGluRs in A549 cells were determined by real-time PCR. Immunohistochemistry was used to analyze the protein expression of mGluR4 and mGluR8 in A549 cells and lung tissue sections obtained from lung adenocarcinoma patients. To observe the effects of mGluR8 and mGluR4 activation on the growth of A549 cells, the cultured cells were treated with (S)-3,4-DCPG (an agonist of mGluR8) and VU0155041 (an agonist of mGluR4), respectively, and then the cell viability was analyzed by CCK-8 kit, the percentage of DNA synthesis was detected by EdU incorporation, and the apoptosis of the cells was measured by hoechst 33258 staining and flow cytometry. The results showed that there were low expressions of mGluR1, mGluR5, mGluR6, mGluR7 mRNA, no expression of mGluR2 and mGluR3 mRNA, and high expressions of mGluR8 and mGluR4 mRNA in A549 cells. Accordingly, there were also mGluR4 and mGluR8 protein expressions in the A549 cells and the lung adenocarcinoma tissue sections. VU0155041 had no effect on the growth of A549 cells, but (S)-3,4-DCPG significantly decreased the cells' growth in a dose-dependent manner and increased the apoptosis of the cells. The results revealed a role of mGluR8 in the growth and apoptosis of A549 cells and suggested a potential target for clinical treatment of lung cancer.
本研究旨在检测代谢型谷氨酸受体(mGluRs)在肺癌A549细胞中的表达,并探讨mGluR8和mGluR4激活对A549细胞体外生长的影响。通过实时PCR测定A549细胞中mGluRs 8种亚型的mRNA表达水平。采用免疫组织化学方法分析mGluR4和mGluR8在A549细胞及肺腺癌患者肺组织切片中的蛋白表达。为观察mGluR8和mGluR4激活对A549细胞生长的影响,分别用(S)-3,4-DCPG(mGluR8激动剂)和VU0155041(mGluR4激动剂)处理培养的细胞,然后用CCK-8试剂盒分析细胞活力,通过EdU掺入检测DNA合成百分比,并用hoechst 33258染色和流式细胞术检测细胞凋亡。结果显示,A549细胞中mGluR1、mGluR5、mGluR6、mGluR7 mRNA表达较低,mGluR2和mGluR3 mRNA无表达,mGluR8和mGluR4 mRNA表达较高。相应地,A549细胞和肺腺癌组织切片中也有mGluR4和mGluR8蛋白表达。VU0155041对A549细胞生长无影响,但(S)-3,4-DCPG以剂量依赖性方式显著降低细胞生长并增加细胞凋亡。结果揭示了mGluR8在A549细胞生长和凋亡中的作用,并提示其可能成为肺癌临床治疗的潜在靶点。