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目的代谢型谷氨酸受体5(mGlu5)在肝组织中具有表达,并且在肝脏的病理过程中发挥重要的调节作用。本课题组以往的研究结果表明,mGlu5的激动剂和阻断剂影响人肝癌细胞系HepG2的生长、凋亡、浸润和转移等功能,提示mGlu5在肝癌的形成和发展中发挥一定的作用。为进一步验证mGlu5在肝癌中的作用,本研究在此基础上选用小鼠肝原代细胞和小鼠来源的肝癌细胞系Hepa1-6为实验模型,通过比较mGlu5在两者中的表达及功能的差异,深入探讨mGlu5影响肝癌的内在机制。方法采用噻唑蓝法、免疫印迹法分别检测了mGlu5对小鼠肝癌细胞活力的影响,两种细胞中mGlu5的表达及mGlu5的功能。结果在小鼠肝原代细胞和肝癌细胞中均有mGlu5的表达,在肝癌细胞中的表达量高于小鼠肝原代细胞;激活Hepa1-6中的mGlu5所需DHPG(mGlu5的激动剂)剂量低于激活小鼠肝原代细胞中mGlu5所需剂量;激活mGlu5促进ERK信号通路激活,在Hepa1-6细胞中,100μmol/L DHPG即可使ERK信号通路激活,而在小鼠肝原代细胞中,激活ERK信号通路需要300μmol/LDHPG。激活mGlu5不影响两种细胞中的JNK以及p38信号通路。激活mGlu5能增加Hepa1-6细胞活力。结论mGlu5在肝细胞和肝癌细胞中的表达及功能存在差异,此差异可能是肝细胞癌发生发展的原因之一。
The purpose of metabotropic glutamate receptor 5 (mGlu5) expression in liver tissue, and in the pathological process of the liver play an important regulatory role. Our previous results showed that mGlu5 agonists and blockers affect the growth, apoptosis, invasion and metastasis of human hepatocellular carcinoma cell line HepG2, suggesting that mGlu5 may play a role in the formation and development of liver cancer. In order to further verify the role of mGlu5 in hepatocellular carcinoma, we selected mouse primary hepatocytes and mouse hepatoma Hepa1-6 cells as experimental models, and compared the expression and function of mGlu5 Differences, in-depth study of mGlu5 affect the intrinsic mechanism of liver cancer. Methods The effect of mGlu5 on the viability of mouse hepatoma cells, the expression of mGlu5 and the function of mGlu5 in two kinds of cells were detected by MTT method and Western blotting respectively. Results The expression of mGlu5 in mouse primary hepatocytes and hepatocellular carcinoma cells was higher than that in mouse primary hepatocytes. The mGlu5-required DHPG (agonist of mGlu5) The dose of mGlu5 was lower than that of mGlu5 in primary mouse liver cells. Activation of mGlu5 promoted the activation of ERK signaling pathway. In Hepa1-6 cells, ERK signaling was activated by 100μmol / L DHPG, In cells, 300μmol / LDHPG is required to activate the ERK signaling pathway. Activation of mGlu5 did not affect both JNK and p38 signaling pathways in both cells. Activation of mGlu5 can increase Hepa1-6 cell viability. Conclusion The expression and function of mGlu5 in hepatocytes and hepatocellular carcinoma cells are different. This difference may be one of the reasons for the development of hepatocellular carcinoma.