表皮生长因子促进子宫内膜腺癌细胞增殖机制研究

发布时间：2018-04-03 13:23

本文选题：子宫内膜肿瘤　切入点：表皮生长因子　出处：《青岛大学》2017年硕士论文

【摘要】：目的:研究表皮生长因子(epidermal growth factor,EGF)对子宫内膜腺癌细胞系Ishikawa增殖的影响,并探讨雌激素受体α(ERα)和Ack1激酶在其调控机制中的作用。方法:(1)无雌激素环境下,EGF和酪氨酸激酶抑制剂Dasatinib(达沙替尼)分别作用于子宫内膜腺癌细胞系Ishikawa后,应用CCK-8法检测子宫内膜腺癌细胞的增殖情况;应用统计学软件SPSS19.0进行数据分析。(2)无雌激素环境下,EGF和酪氨酸激酶抑制剂Dasatinib(达沙替尼)分别作用于子宫内膜腺癌细胞系Ishikawa后,应用Western blot法检测Ishikawa细胞ERα及Ack1激酶非磷酸化与磷酸化状态蛋白的表达情况;应用Image J图像分析软件,通过磷酸化目的蛋白与非磷酸化目的蛋白条带的吸光度值比值进行目的蛋白的相对含量分析。结果:(1)在无雌激素条件下,EGF(10μg/L)能够促进子宫内膜腺癌Ishikawa细胞增殖。与对照组比较有显著性差异,P0.05。(2)达沙替尼(10 nmol/L-800 nmol/L)呈剂量依赖性抑制EGF诱导的子宫内膜腺癌Ishikawa细胞增殖。与EGF处理组比较有显著性差异,P0.05。(3)EGF能够诱导Ishikawa细胞Ack1激酶及ERαTyr-537特异位点磷酸化。与对照组相比,Ack1激酶和ERαTyr-537位点的磷酸化水平升高,结果有显著性差异,P0.05。(4)达沙替尼能够抑制Ishikawa细胞EGF诱导的Ack1激酶及ERαTyr-537磷酸化。当达沙替尼浓度大于400 nmol/L时,Ack1激酶及ERαTyr-537位点磷酸化被完全抑制,与对照组比较有显著性差异,P0.05。结论:(1)EGF促进子宫内膜腺癌Ishikawa细胞Ack1激酶及ERαTyr-537磷酸化及促进Ishikawa细胞增殖(P0.05),EGF在子宫内膜癌细胞发生发展作用明显,为寻找新型抗肿瘤药物治疗靶点提供了理论基础。(2)达沙替尼能够抑制子宫内膜腺癌Ishikawa细胞EGF诱导的Ack1激酶及ERαTyr-537磷酸化,并抑制Ishikawa细胞增殖(P0.05),达沙替尼可应用于子宫内膜癌的治疗。
[Abstract]:Aim: to investigate the effects of epidermal growth factor (EGF) on the proliferation of endometrial adenocarcinoma cell line Ishikawa, and to explore the role of estrogen receptor alpha (ER 伪) and Ack1 kinase in its regulatory mechanism.Methods EGF and Dasatinib (Dasatinib), an inhibitor of tyrosine kinase, were used to detect the proliferation of endometrial adenocarcinoma cell line Ishikawa by CCK-8.Statistical software SPSS19.0 was used to analyze the data. (2) EGF and Dasatinib (Dasatinib), a tyrosine kinase inhibitor, were treated on endometrial adenocarcinoma cell line Ishikawa respectively.The expression of ER 伪 and Ack1 kinase non-phosphorylated and phosphorylated state proteins in Ishikawa cells was detected by Western blot assay, and Image J image analysis software was used.The relative content of the target protein was analyzed by the ratio of the absorbance of the phosphorylated target protein to the non-phosphorylated target protein band.Results 10 渭 g 路L ~ (-1) of Ishikawa could promote the proliferation of endometrial adenocarcinoma Ishikawa cells without estrogen.Compared with the control group, there was a significant difference (P0.05.22) Dasatinide (10 nmol/L-800 nmol / L) inhibited the proliferation of endometrial adenocarcinoma Ishikawa cells induced by EGF in a dose-dependent manner.Compared with the EGF treatment group, there was a significant difference between the two groups (P 0.05. 05. 3) EGF could induce the phosphorylation of Ack1 kinase and ER 伪 Tyr-537 specific sites in Ishikawa cells.Compared with the control group, the phosphorylation of Ack1 kinase and ER 伪 Tyr-537 sites was significantly increased. The results showed that Dasatinib could inhibit Ack1 kinase and ER 伪 Tyr-537 phosphorylation induced by EGF in Ishikawa cells.The phosphorylation of Ack1 kinase and ER 伪 Tyr-537 sites was completely inhibited when the concentration of dasatinib was more than 400 nmol/L, which was significantly different from that of the control group (P 0.05).Conclusion the Ack1 kinase and ER 伪 Tyr-537 phosphorylation of endometrial adenocarcinoma Ishikawa cells and the proliferation of Ishikawa cells can be significantly promoted by the growth of EGF in endometrial carcinoma cell line P0.05EGF may play an important role in the carcinogenesis and development of endometrial carcinoma cells.Dasatinib can inhibit Ack1 kinase and ER 伪 Tyr-537 phosphorylation induced by EGF in endometrial adenocarcinoma Ishikawa cells.The proliferation of Ishikawa cells was inhibited by P0.05, and dassatinide was used in the treatment of endometrial carcinoma.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.33

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