Hedgehog信号通路中Gli1和Foxm1在卵巢癌细胞株Skov3中的表达及其意义

发布时间：2018-04-09 03:11

  本文选题：卵巢癌　切入点：Hedgehog信号通路　出处：《河北医科大学》2017年硕士论文

【摘要】：目的:卵巢癌是严重威胁女性生命健康的恶性肿瘤,死亡率居妇科恶性肿瘤之首。因为卵巢深居盆腔,又缺乏诊断卵巢癌的高度敏感指标,导致60%—70%患者确诊时已属晚期。因此,卵巢癌的诊断及预后一直是妇科临床工作者研究的重点和热点。近年来Hedgehog信号通路在恶性肿瘤中的作用已成为研究热点,Hedgehog信号通路的异常激活与肿瘤的发生发展密切相关,参与调控恶性肿瘤细胞的发生、发展、增殖及侵袭转移过程。本实验采用细胞培养、MTT法、实时荧光定量PCR、Western blot等方法检测添加Hedgehog信号通路激动剂—Shh多肽后,通路转录因子Gli1及其下游靶基因Foxm1在卵巢癌细胞株Skov3中的表达变化及其细胞生长变化,探讨Hedgehog信号通路中Gli1和Foxm1在卵巢癌细胞株Skov3中的表达情况及其对卵巢癌细胞Skov3增殖的影响,为临床卵巢癌的早期诊断和治疗提供新的理论基础。方法:1培养卵巢癌Skov3细胞株。2(1)实验组:添加Hedgehog信号通路激动剂Shh(Human Sonic Hedgehog)多肽(200ng/ml);(2)对照组:不添加Hedgehog信号通路激动剂Shh多肽。3应用MTT法检测各组卵巢癌Skov3细胞在24h、48h、72h的增殖情况。4采用实时荧光定量PCR法检测各组卵巢癌Skov3细胞中24h、48h、72h Gli1m RNA和Foxm1m RNA的表达情况。5 Western blot法检测各组卵巢癌Skov3细胞中24h、48h、72h Gli1和Foxm1蛋白表达的情况。6用SPSS21.0统计软件对所有数据进行统计学分析。所有数据为正态分布,以均数±标准差((?)±s)方式表示,两组间均数比较用t检验,P0.05为有统计学意义。结果:1 MTT法检测各组细胞增殖情况:24、48、72小时,实验组较对照组细胞增殖速度明显加快,统计学上有显著差异(P0.05)。2实时荧光定量PCR法检测24h、48h、72h两组细胞中Gli1m RNA和Foxm1m RNA的表达情况:实验组Gli1m RNA和Foxm1m RNA在24h、48h、72h的表达量与对照组相比均明显升高,统计学上有显著差异(P0.05)。3 Western blot法检测24h、48h、72h两组细胞中Gli1和Foxm1蛋白表达情况:实验组与对照组相比,在24h、48h、72h Gli1和Foxm1蛋白表达均明显升高,统计学上有显著差异(P0.05)。结论:1 Hedgehog信号传导通路激动剂Shh多肽可增加通路活性(Gli活性),促进卵巢癌细胞生长。2 Hedgehog信号传导通路,通过高调Foxm1的表达来促进卵巢癌细胞增长。
[Abstract]:Objective: ovarian cancer is a serious threat to women's life and health, the mortality rate of gynecological malignancies is the first.Because the ovary is deep in the pelvic cavity and lacks the highly sensitive index to diagnose the ovarian cancer, 60-70% of the patients are in the late stage by the time of diagnosis.Therefore, the diagnosis and prognosis of ovarian cancer has been the focus and focus of gynecological clinicians.In recent years, the role of Hedgehog signaling pathway in malignant tumors has become a hot topic. The abnormal activation of Hedgehog signaling pathway is closely related to the occurrence and development of tumor. It is involved in the process of tumor cell development, development, proliferation, invasion and metastasis.In this experiment, the Hedgehog signal pathway agonist -Shh polypeptide was detected by cell culture MTT assay and real-time fluorescence quantitative PCR Western blot.Expression and growth of transcriptional factor Gli1 and its downstream target gene Foxm1 in ovarian cancer cell line Skov3.To investigate the expression of Gli1 and Foxm1 in the Hedgehog signaling pathway in ovarian cancer cell line Skov3 and its effect on the proliferation of ovarian cancer cell line Skov3, and to provide a new theoretical basis for early diagnosis and treatment of ovarian cancer.Methods the Skov3 cell line of ovarian cancer was cultured with 1: 1) experimental group: the control group was supplemented with Shh(Human Sonic Hedgehog) polypeptide (200ng / ml ~ (2)). The increase of Skov3 cells in each group was detected by MTT assay at 24 h, 48 h and 72 h, without the addition of Hedgehog signal pathway agonist Shh 3.The expression of Gli1m RNA and Foxm1m RNA in ovarian cancer Skov3 cells was detected by real-time fluorescence quantitative PCR method. 5. 5 Western blot assay was used to detect the expression of Gli1 and Foxm1 protein at 24 h, 48 h and 72 h by SPSS21.0 statistical software.All the data were analyzed statistically.All the data were normal distribution in the form of mean 卤standard deviation (卤s). The comparison between the two groups was statistically significant with t test (P0.05).Results the cell proliferation of each group was detected by 1: 1 MTT assay for 72 hours. The proliferation rate of the experimental group was significantly faster than that of the control group.There was statistically significant difference in the expression of Gli1m RNA and Foxm1m RNA between the two groups by real-time fluorescence quantitative PCR. The expression of Gli1m RNA and Foxm1m RNA in the experimental group was significantly higher than that in the control group.There was significant difference in the expression of Gli1 and Foxm1 protein between the two groups by P0.05.3 Western blot method. Compared with the control group, the expression of Gli1 and Foxm1 in the experimental group was significantly higher than that in the control group at 48 h and 72 h, and there was a significant difference between the two groups in the expression of Gli1 and Foxm1 protein (P 0.05).Conclusion Shh polypeptide can increase the activity of Hedgehog signal transduction pathway and promote the growth of ovarian cancer cell line 2. 2 Hedgehog signal transduction pathway, and promote the growth of ovarian cancer cells through the expression of high intensity Foxm1.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.31

【参考文献】

相关期刊论文 前9条

1 陈丹;陈红;董良波;杨红;王晶晶;;Gli1与FoxM1在子宫颈癌组织和细胞中的表达及意义[J];武汉大学学报(医学版);2016年03期

2 王艳;李刚;佐满珍;;FoxM1剪接异构体在上皮性卵巢癌中的表达及临床意义[J];河北医药;2016年04期

3 郑建涛;张琳;李良庆;;Gli1及Foxm1水平对胃癌转移状况及治疗预后的影响[J];胃肠病学和肝病学杂志;2014年12期

4 刘静波;马玲;;卵巢癌Hedgehog通路研究进展[J];实用癌症杂志;2014年04期

5 刘道永;邓军;郭伟;张明亮;陈勇;;Gli-1蛋白在胃癌中的表达及临床意义[J];中华全科医学;2013年09期

6 应倩;夏庆民;郑荣寿;张思维;陈万青;;中国2009年宫颈癌发病与死亡分析[J];中国肿瘤;2013年08期

7 覃利菊;;卵巢癌诊断和治疗的现状及进展[J];中外医学研究;2013年19期

8 陈国庆;姚珍薇;罗欣;;转录因子FOXM1在上皮性卵巢癌中的表达及临床意义[J];生命科学研究;2011年01期

9 黄海林;胡志前;王为民;王毅;蔡清萍;王强;;Hedgehog信号传导通路在乳腺癌中表达增强[J];基础医学与临床;2009年05期

相关硕士学位论文 前2条

1 王俐红;Shh和Foxm1在上皮性卵巢肿瘤组织中的表达及临床意义[D];河北医科大学;2015年

2 赵欢;HHIP基因在子宫内膜癌中甲基化状态及表达[D];河北医科大学;2011年

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