γ-氨基丁酸对胆管癌细胞株QBC939增殖和侵袭转移的影响

Effects of gamma-aminobutyric acid on the proliferation and invasion of cholangiocarcinoma cell line QBC939

目的观察γ-氨基丁酸（GABA）对胆管癌细胞株QBC939增殖、凋亡和侵袭转移的影响。方法取GABA母液，用含10％胎牛血清的1640培养液稀释，配成浓度为1、10、100、1000umol／L为实验组；对照组仅加入与实验组相同体积的完全培养液。采用MTT比色法观察GABA对人胆管癌细胞株QBC939增殖的作用，流式细胞仪检测GABA对胆管癌细胞株QBC939凋亡的影响，Transwell小室分析GABA对胆管癌细胞侵袭能力的影响，明胶酶谱法分析GABA对胆管癌细胞株QBC939分泌的基质金属蛋白酶（MMP）活性的影响，放射免疫分析法检测cAMP含量。采用单因素方差分析。结果GABA浓度由1umol／L增至1000umol／L，GABA对胆管癌细胞的抑制率由2．6％增至26．8％；细胞凋亡率由4．80％±0．04％增至28．03％±0．01％；穿透Matrigel胶的能力由（60±10）个减至（43±4）个，均呈剂量依赖性（F=7．833，83．765，7．598，P〈0．05）。同时胆管癌细胞分泌的MMP-2和MMP-9活性下降，cAMP含量增加，与对照组比较差异有统计学意义（F=9．507，9．148，27．418，P〈0．05）。结论GABA可能通过受体后信息介导途径，诱导细胞凋亡，并减弱胆管癌细胞株QBC939的侵袭转移能力，可能与其抑制基质蛋白酶MMP-2和MMP-9的活性有关。

Objective To explore the effects of gamma-aminobutyric acid （GABA） on the proliferation, apoptosis and invasion of cholangiocarcinoma cell line QBC939. Methods Cholangiocarcinoma cell line QBC939 was cultured via routine methods, and was treated with different concentrations of GABA （1, 10, 100, 1000 umol/L, test group） or with complete culture medium （ control group）. The effects of GABA on the proliferation, apoptosis and invasion of cholangiocarcinoma cell line QBC939 were investigated by MTT assay, flow cytometry and Transwell cell culture chamber assay, respectively. The effects of GABA on the activity of matrix metalloproteinase （MMP） secreted by cholangiocarcinoma cells were analyzed by gelatin zymogram, and the content of intracellular cyclic adenosine monophosphate （cAMP） was measured by radioimmunoassay. All data were analyzed by one-way ANOVA. Results As the GABA concentration increased from 1umol/L to 1000 umoL/L, the inhibition rate of GABA on cholangioeareinoma cell line QBC939 was increased from 2.6% to 26.8% , and the apoptosis rate also increased from 4.80% ± 0.04% to 28.03% ± 0.01%. The number of cholangiocarcinoma cells that migrated through the Matrigel gel decreased from 60 ± 10 to 43 ± 4, in a dose-dependent manner （F = 7. 883, 83. 765, 7. 598, P 〈0.05）. The activity of MMP-2 and MMP-9 was decreased. The content of intracellular cAMP was increased, and there was significant difference in the content of intracellular cAMP between test and control groups （F = 9. 507, 9. 148, 27. 418, P 〈 0.05）. Conclusions GABA can inhibit the proliferation and invasion of cholangiocarcinoma cell line QBC939 ceils by promoting the apoptosis and inhibiting the activity of MMP-2 and MMP-9. The process may be mediated by the information transmission of post-receptor.