目的 研究糖原合酶激酶-3β抑制剂4,6-二取代吡咯并嘧啶(TWS119)体外抗肿瘤活性和对人γδT细胞活性的影响。方法 用不同浓度的4,6-二取代吡咯并嘧啶分别作用于人胃癌SGC-7901细胞和γδT细胞24、48和72 h后,用CCK-8法测定4,6-二取代吡咯并嘧啶对人胃癌SGC-7901细胞和γδT细胞增殖的影响及γδT细胞对SGC-7901细胞杀伤能力;流式细胞术分析4,6-二取代吡咯并嘧啶对SGC-7901细胞凋亡的影响,Western blot检测4,6-二取代吡咯并嘧啶对Bcl-2、p-STAT3和p-ERK1/2表达的影响。结果 0~8.0 μmol·L-1 4,6-二取代吡咯并嘧啶能抑制人胃癌SGC-7901细胞的生长并诱导其凋亡,抑制Bcl-2的表达,促进p-ERK1/2和p-STAT3的表达。而在γδT细胞中,4,6-二取代吡咯并嘧啶在0~4.0 μmol·L-1时能促进其增殖和对SGC-7901细胞的体外杀伤活性,促进Bcl-2的表达,抑制p-ERK1/2和p-STAT3的表达。结论 一定浓度的4,6-二取代吡咯并嘧啶能抑制人胃癌SGC-7901细胞的增殖并诱导其凋亡,同时能促进γδT细胞增殖和对SGC-7901细胞的杀伤活性,其机制可能与4,6-二取代吡咯并嘧啶激活Wnt信号传导通路及其对p-ERK1/2、Bcl-2和p-STAT3表达的影响有关。
Abstract
OBJECTIVE To study the antitumor activity of TWS119 and its effect on the killing activity of γδT cells in vitro. METHODS CCK-8 assay was used for detecting the influence of TWS119 on the proliferation of SGC-7901 and γδT cells and the cytotoxic activity of γδT cells to SGC-7901 cells. The cell apoptosis was measured by flow cytometric analysis.The protein expression was examined by Western blot analysis.RESULTS It was found that TWS119 could inhibit the growth and induce the apoptosis of SGC-7901 cells.The expression of Bcl-2 in SGC-7901 cells treated with TWS119 was downregulated,while p-ERK1/2 and p-STAT3 were upregulated.In addition, TWS119 at 0.5-4.0 μmol·L-1 could promote the growth of γδT cells and enhance the cytotoxic activity of γδT cells to SGC-7901 cells especially at 4.0 μmol·L-1 for 72 h. Furthermore, TWS119 could upregulate the expression of Bcl-2 and inhibit the expression of p-ERK1/2 and p-STAT3.CONCLUSION TWS119 in some concentrations can inhibit the growth of SGC-7901 cells, promote the γδT cells growth and enhance the cytotoxic activity of γδT cells to SGC-7901 cells, and the mechanism may be involve in Wnt, ERK1/2, Bcl-2 and p-STAT3 signaling pathways.
关键词
4,6-二取代吡咯并嘧啶 /
γδT细胞 /
SGC-7901 /
细胞增殖 /
凋亡
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Key words
TWS119 /
γδT cells /
SGC-7901 /
cell proliferation /
apoptosis
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中图分类号：
R392.12
Q253
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参考文献
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脚注
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基金
南京军区医学科技创新研究基金资助项目(14MS032)
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