[1]霍浩然,秦瑞峰,薛佳栋,等.基于PI3K/Akt/mTOR通路探讨川芎嗪诱导人胃癌细胞凋亡和自噬作用的机制研究[J].西部中医药,2022,35(10):12-16.[doi:10.12174/j.issn.2096-9600.2022.10.04]
　HUO Haoran,QIN Ruifeng,XUE Jiadong,et al.Research on the Mechanism of TMP- induced Apoptosis and Autophagy in Human Gastric Cancer Cells Based on P13k/Akt/mTOR Pathway[J].Western Journal of Traditional Chinese Medicine,2022,35(10):12-16.[doi:10.12174/j.issn.2096-9600.2022.10.04]
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基于PI3K/Akt/mTOR通路探讨川芎嗪诱导人胃癌细胞凋亡和自噬作用的机制研究
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《西部中医药》[ISSN:2096-9600/CN:62-1204/R]

卷:
35
期数:
2022年10期
页码:
12-16
栏目:
理论论著
出版日期:
2022-10-15

文章信息/Info

Title:
Research on the Mechanism of TMP- induced Apoptosis and Autophagy in Human Gastric Cancer Cells Based on P13k/Akt/mTOR Pathway
作者:
霍浩然, 秦瑞峰, 薛佳栋, 袁增江
邯郸市中心医院，河北 邯郸 056001
Author(s):
HUO Haoran, QIN Ruifeng, XUE Jiadong, YUAN Zengjiang
Handan Central Hospital, Handan 056001, China
关键词:
胃癌; 川芎嗪; PI3K/Akt/mTOR; 凋亡; 自噬
Keywords:
gastric cancer; tetramethylpyrazine; PI3K/Akt/mTOR; apoptosis; autophagy
分类号:
R735
DOI:
10.12174/j.issn.2096-9600.2022.10.04
文献标志码:
A
摘要:
目的研究川芎嗪（tetramethylpyrazine，TMP）对人胃癌SGC-7901细胞凋亡和自噬的影响并探索其分子机制。 方法分别以二甲基亚砜（dimethyl sulfoxide，DMSO）（空白对照组）、TMP（200、400、800 μg/mL）和LY294002（5 μg/mL）干预人胃癌SGC-7901细胞，48 h后通过MTT法检测细胞增殖抑制率，Annexin V-FITC/PI流式细胞术检测细胞凋亡水平，观察细胞自噬，Western blot法检测磷酸化磷酸肌醇3激酶（phosphorylated phosphatidylinositol 3-kinase，P-PI3K）、磷酸化蛋白激酶B（phosphorylated protein kinase B，p-Akt）、磷酸化哺乳动物雷帕霉素靶蛋白（phosphorylated mammalian target of rapamycin，p-mTOR）、活化型半胱氨酸蛋白酶9（cleaved cysteinyl aspartate specific proteinase 9，Cleaved Caspase-9）、活化型半胱氨酸蛋白酶3（cleaved cysteinyl aspartate specific proteinase 3，Cleaved Caspase-3）、微管相关蛋白1轻链3（microtubule-associated protein 1 light chain 3，LC3）蛋白表达，计算LC3-Ⅱ/LC3-Ⅰ比值。 结果TMP 200、400、800 μg/mL或LY294002 5 μg/mL可明显提高细胞增殖抑制率、凋亡指数和自噬溶酶体数量。TMP 400、800 μg/mL或PI3K特异性抑制剂LY294002 5 μg/mL可明显降低p-PI3K、p-Akt、p-mTOR相对表达量，提高Cleaved Caspase-9、Cleaved Caspase-3相对表达量和LC3-Ⅱ/LC3-Ⅰ比值，差异有统计学意义（P＜0.05）。TMP 800 μg/mL组对细胞增殖抑制，促其凋亡和自噬，p-PI3K、p-Akt、p-mTOR、Cleaved Caspase-3表达及LC3-Ⅱ/LC3-Ⅰ比值的影响均明显优于LY294002 5 μg/mL组（P＜0.05或P＜0.01）。 结论TMP可能通过抑制PI3K/Akt/mTOR通路活化而诱导人胃癌细胞凋亡和自噬。
Abstract:
ObjectiveTo study the effects of TMP on the apoptosis and autophagy in human gastric cancer SGC-7901 cells and explore its molecular mechanism. MethodsDMSO (blank control group), TMP (200, 400 and 800 μg/mL) and LY294002(5 μg/mL) were used to intervene human gastric cancer SGC-7901 cells, the inhibition rate of cell proliferation was measured via MTT method in 48 hours, the levels of cellular apoptosis was detected by using Annexin V-FITC/P1 flow cytometry (FCM), to observe cellular autophagy, the expressions of p-PI3K, p-Akt, p-mTOR, Cleaved Caspase-9, Cleaved Caspase-3 and LC3 proteins were detected by using Western blot method, to calculate LC3-Ⅱ/LC3-Ⅰ ratio. ResultsTMP in the concentrations of 200, 400 and 800 μg/mL or LY294002 in the concentration of 5 μg/mL could notably raise the inhibition rate of cell proliferation, apoptotic index and the number of autophagic lysosomes. TMP in the concentrations of 400 and 800 μg/mL or PI3K specific inhibitor LY294002 in the concentration of 5 μg/mL could remarkably decrease the relative expressions of p-PI3K, p-Akt and p-mTOR, increase the relative expressions of Cleaved Caspase-9, Cleaved Caspase-3 and LC3-Ⅱ/LC3-Ⅰ ratio, and the difference had statistical meaning (P<0.05). TMP group of 800 μg/mL demonstrated the more noticeable effects on the inhibition of cellular proliferation, promoting the apopotosis and autophagy of the cells, the expressions of p-PI3K, p-Akt, p-mTOR, Cleaved Caspase-3 and LC3-Ⅱ/LC3-Ⅰ ratio compared with the effects of LY294002 group of 5 μg/mL (P<0.05 or P<0.01). ConclusionTMP could induce the apoptosis and autophagy of human gastric cancer cells possibly through inhibiting the activation of PI3K/Akt/mTOR pathway.

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备注/Memo

备注/Memo:
霍浩然（1984—），男，硕士学位，副主任医师。研究方向：普外科疾病的研究。河北省医学科学研究重点课题计划（20181824）。
更新日期/Last Update: 2022-10-15