Third Affiliated Hospital of Southern Medical University, Guangzhou, China.
Adv Clin Exp Med. 2022 Aug;31(8):881-888. doi: 10.17219/acem/130445.
Multiple myeloma (MM) is a malignant plasma cancer which remains difficult to be cured. Recently, numerous research studies have appeared, exploring MM from molecular level. However, there is no study about the impact of metabotropic glutamate receptors (mGluRs), especially mGluR5, on MM progression. Thus, the present research was dedicated to the exploration of the influence of mGluR5 on MM.
In this research, we used quantitative real-time polymerase chain reaction (qRT-PCR) to check the gene expression in MM, western blot assay to check the protein expression of the gene, MTT assay to quantify the cell viability, and flow cytometry (FCM) apoptosis method to evaluate cell apoptosis in order to acquire the results. The purpose was to assess the role of mGluR5 in MM cells.
The qRT-PCR was used and it was found that mGluR5 was overexpressed in MM cell lines and MM tissues compared to normal ones. To better observe the function of mGluR5 in MM, cell viability and apoptosis were checked using MTT and FCM apoptosis assays after the treatment with agonists and antagonists.
Agonist-induced mGluR5 upregulation could promote MM cell viability and inhibit apoptosis. The same results were obtained through MTT and FCM apoptosis assays after upregulation and downregulation of mGluR5 by transfection. To further investigate the inner mechanism, the effect of mGluR5 on Ras-MAPK pathway was checked using western blot. It was found that the upregulation of mGluR5 could activate the Ras-MAPK pathway.
The mGluR5 might be involved in promoting cell proliferation and inhibiting cell apoptosis in MM. It can be an essential biomarker in the screening for MM and a potential part of future MM therapies.
多发性骨髓瘤（MM）是一种难以治愈的恶性浆细胞癌。最近，许多研究从分子水平上探讨了 MM。然而，目前还没有研究涉及代谢型谷氨酸受体（mGluRs），特别是 mGluR5，对 MM 进展的影响。因此，本研究旨在探讨 mGluR5 对 MM 的影响。
在这项研究中，我们使用定量实时聚合酶链反应（qRT-PCR）检查 MM 中的基因表达，使用蛋白质印迹法（western blot assay）检查基因的蛋白表达，使用 MTT 测定法（MTT assay）量化细胞活力，使用流式细胞术（FCM）凋亡法评估细胞凋亡，以获得结果。目的是评估 mGluR5 在 MM 细胞中的作用。
使用 qRT-PCR 发现 mGluR5 在 MM 细胞系和 MM 组织中过度表达，与正常组织相比。为了更好地观察 mGluR5 在 MM 中的作用，在用激动剂和拮抗剂处理后，使用 MTT 和 FCM 凋亡测定法检查细胞活力和凋亡。
激动剂诱导的 mGluR5 上调可促进 MM 细胞活力并抑制凋亡。通过转染上调和下调 mGluR5 后，通过 MTT 和 FCM 凋亡测定法获得了相同的结果。为了进一步研究内在机制，使用 western blot 检查了 mGluR5 对 Ras-MAPK 途径的影响。结果发现，mGluR5 的上调可激活 Ras-MAPK 途径。
mGluR5 可能参与促进 MM 中的细胞增殖和抑制细胞凋亡。它可以作为 MM 筛查的重要生物标志物，也是未来 MM 治疗的潜在部分。
