氯化锂对人子宫内膜癌HEC-1A细胞增殖及Shh表达的影响
Effects of Lithium Chloride on the Proliferation of Human Endometrial Carcinoma Cell Line HEC-1A and Shh Expression
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摘要:目的 研究Wnt、Shh在人子宫内膜癌细胞HEC-1A中的表达, 应用不同浓度糖原合成酶激酶-3的抑制剂氯化锂(LiCl)处理, 观察对人子宫内膜癌细胞HEC-1A体外增殖和Shh表达的影响。方法 体外培养人子宫内膜癌细胞HEC-1A, 不同浓度LiCl处理细胞6~48 h, 利用免疫细胞化学法测定HEC-1A细胞上Wnt、Shh表达; MTT比色法检测HEC-1A细胞增殖活性; RT-PCR检测核转录因子Gli-1的表达水平。结果 Wnt、Shh在HEC-1A细胞上有表达; 不同浓度LiCl处理后, Shh阳性细胞先出现增殖, 24 h时细胞量达到高峰, 随着时间延长, Shh细胞量减少, 高浓度LiCl组在48h时细胞减少最显著(P < 0.05);MTT检测细胞增殖结果显示LiCl对HEC-1A细胞的生长有抑制作用, 且呈浓度、时间依赖性。RT-PCR检测结果显示Gli-1基因的表达在24 h组水平最高, 与Shh的阳性细胞变化情况相符合。结论 Wnt信号可以调控Shh的表达, LiCl能抑制HEC-1A细胞的增殖, 并具有浓度、时间依赖趋势。Abstract:Objectives To study the expression of the wnt and sonic hedgehog homolog(Shh) proteins in the endometrial carcinoma cell line HEC-1A;and to study the growth of HEC-1A cells treated with lithium chloride, an inhibitor of glycogen synthase kinase-3.Methods The HEC-1A cells were cultivated in vitro.Immunofluorescence was used to determine the expression of wnt and Shh in the cells. 3-(4, 5 - Dimethylthiazol - 2 - yl) -2, 5-diphenyltetrazolium bromide(MTT) was used to investigate the proliferation of HEC-1A cells treated with different concentrations of lithium chloride.In addition, real-time polymerase chain reaction(RT - PCR) was used to determine the expression level of the nuclear transcription factor Gli-1 -Results Wnt and Shh were expressed in the HEC-1A cells.The amount of Shh initially increased, and then decreased.Lithium chloride inhibited the growth of HEC-1A cells in a concentration-dependent manner.The RT-PCR results showed that the highest expression level of Gli-1 was in the 24 h group, which corresponded to a change in the Shh level.Conclusion The wnt signaling pathway can regulate the Shh expression.Moreover, lithium chloride can inhibit the proli feration of HEC-1A cells in a concentration-time-dependent manner.
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