摘要
目的研究大黄素衍生物A对具有不同淋巴结转移潜能的卵巢癌细胞的迁移和侵袭能力的抑制作用,并且探讨其可能的作用机制。方法四甲基偶氮唑蓝(MTT)法检测不同浓度大黄素衍生物A处理人卵巢浆液性乳头状腺癌细胞(SKOV3)和具有淋巴结定向高转移能力的亚克隆细胞(SKOV3-pm4)24 h后细胞的增殖作用。免疫荧光法观察细胞中Rac1蛋白的分布情况。不同浓度药物作用两种细胞后,Western blot检测细胞Rac1蛋白的表达量,细胞划痕实验检测细胞的迁移能力,Transwell侵袭实验测定细胞的侵袭能力。结果在一定浓度范围内大黄素衍生物A对SKOV3和SKOV3-pm4细胞的增殖均有抑制作用,24 h的半数抑制浓度(IC50)分别为23.21和34.46 mg/L。免疫荧光观察到两种细胞中Rac1蛋白均广泛分布于细胞质中,Western blot结果显示SKOV3-pm4细胞Rac1蛋白表达量高于SKOV3(P<0.05)。4和8 mg/L处理组中Rac1蛋白表达较空白组明显降低。细胞划痕实验和Transwell小室侵袭实验结果显示,大黄素衍生物A处理后两种细胞的迁移和侵袭能力均受到抑制。结论大黄素衍生物A能抑制具有高淋巴结转移潜能的卵巢癌细胞的迁移和侵袭能力,其作用机制可能与下调Rac1蛋白的表达有关。
[ Objective ] To investigate the inhibitory effects of Emodin Derivant A on migration and invasion abil- ities of ovarian cancer cells with different lymph node metastatic potential and to explore its possible mechanism. [ Methods ] The cell proliferation of ovarian cancer cells SKOV3 and ovarian cancer cells with highly directional lymphatic metastasis SKOV3-pm4 was tested by MTT assay after treatment with different concentrations of Emodin Derivant A for 24 hours. Immunofluorescence （IF） staining was performed to observe the distribution of Racl. The expression of Racl protein was detected by Western blot. The migration and invasive ability of SKOV3 and SKOV3- pm4 were measured by Scratch assay and Transwell chamber assay. [ Results ] Emodin Derivant A can inhibit the proliferation of the two types of cells within a certain concentration range. The half maximal inhibitory concentration （IC50） at 24 h were 23.21 mg/L and 34.46 mg/L, respectively. Racl protein was widely distributed in the cytoplasm of the two types of cells. Compared with SKOV3 cell, the expression level of Racl in SKOVa-pm4 cell was higher （P 〈 0.05）. The Racl protein expression level after treatment with 4 mg/L and 8 mg/L of Emodin Derivant A was lower than that in the control group. Our studies found that Emodin Derivant A significantly inhibited the invasive and mi- gratory characteristics of SKOV3 and SKOV3-pm4 after 24 h incubation. [ Conclusions ] Emodin Derivant A can in-hibit migration and invasion of ovarian cancer cells with high lymph node metastatic potential and its mechanism might be related to down-regulating the expression of Racl protein.
出处
《中国现代医学杂志》
CAS
北大核心
2015年第19期1-6,共6页
China Journal of Modern Medicine
基金
国家自然基金(No:81060218
81360502)
广西自然科学基金(No:2014GXNSFAA118161)
广西区域性高发肿瘤早期防治研究重点实验室自主研究项目(No:GK2014-ZZ15)
