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本文旨在检测谷氨酸促代谢型受体(metabotropic glutamate receptors,mGluR)各亚型在肺腺癌A549细胞中的表达,并进一步探讨呈高表达的mGluR8和mGluR4激活对A549细胞体外生长的影响。采用real-time PCR技术检测A549细胞mGluR各亚型的m RNA表达,采用免疫组化技术检测A549细胞以及肺腺癌组织mGluR8和mGluR4蛋白表达。采用CCK-8试剂盒检测细胞生长,EdU掺入检测细胞DNA合成率,hoechst 33258染色及流式细胞仪检测细胞的凋亡的变化,分别观察mGluR8的激动剂(S)-3,4-DCPG及mGluR4的激动剂VU0155041对A549细胞生长的影响。结果显示:A549细胞I组促代谢型受体mGluR1和mGluR5m RNA呈低水平表达,II组促代谢型受体mGluR2和mGluR3 mRNA无表达,III组促代谢型受体mGluR4、mGluR6、mGluR7和mGluR8 m RNA均有表达,其中以mGluR8的m RNA表达水平最高,mGluR4表达次之;A549细胞及部分患者肺腺癌组织上有mGluR4和mGluR8的蛋白表达。VU0155041对A549细胞的体外生长无明显影响,而(S)-3,4-DCPG呈剂量依赖性抑制A549细胞的生长,并促进其凋亡。这些结果揭示mGluR8激活具有抑制肺癌细胞生长作用,为肺癌防治的研究提供新的线索。
This article aims to detect the expression of metabotropic glutamate receptors (mGluR) subtypes in lung adenocarcinoma A549 cells and further investigate the effect of mGluR8 and mGluR4 overexpression on A549 cell growth in vitro . Real-time PCR was used to detect m RNA expression in mGluR subtypes of A549 cells. The expression of mGluR8 and mGluR4 in A549 cells and lung adenocarcinoma was detected by immunohistochemistry. The cell growth was detected by CCK-8 kit, DNA synthesis rate was detected by EdU incorporation, the apoptosis of cells was detected by hoechst 33258 staining and flow cytometry. The effects of mGluR8 agonist (S) -3,4-DCPG And mGluR4 agonist VU0155041 on A549 cell growth. The results showed that the expression of mGluR1 and mGluR5mRNA in the A549 cells was low, while the mRNAs of the metabotropic receptors mGluR2 and mGluR3 were not expressed in the A549 cells. The expression of the mGluR4, mGluR6, mGluR7 and mGluR8 m RNA was expressed in all of them, among them, mGluR8 had the highest expression of m RNA and mGluR4, followed by mGluR4 and mGluR8 in A549 cells and some lung adenocarcinoma tissues. VU0155041 had no significant effect on the growth of A549 cells in vitro, while (S) -3,4-DCPG inhibited the growth of A549 cells in a dose-dependent manner and promoted its apoptosis. These results reveal that mGluR8 activation can inhibit the growth of lung cancer cells and provide new clues for the prevention and treatment of lung cancer.