本文選題：卵巢癌　切入點(diǎn)：Hedgehog信號通路　出處：《河北醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:卵巢癌是嚴(yán)重威脅女性生命健康的惡性腫瘤,死亡率居?jì)D科惡性腫瘤之首。因?yàn)槁殉采罹优枨?又缺乏診斷卵巢癌的高度敏感指標(biāo),導(dǎo)致60%—70%患者確診時已屬晚期。因此,卵巢癌的診斷及預(yù)后一直是婦科臨床工作者研究的重點(diǎn)和熱點(diǎn)。近年來Hedgehog信號通路在惡性腫瘤中的作用已成為研究熱點(diǎn),Hedgehog信號通路的異常激活與腫瘤的發(fā)生發(fā)展密切相關(guān),參與調(diào)控惡性腫瘤細(xì)胞的發(fā)生、發(fā)展、增殖及侵襲轉(zhuǎn)移過程。本實(shí)驗(yàn)采用細(xì)胞培養(yǎng)、MTT法、實(shí)時熒光定量PCR、Western blot等方法檢測添加Hedgehog信號通路激動劑—Shh多肽后,通路轉(zhuǎn)錄因子Gli1及其下游靶基因Foxm1在卵巢癌細(xì)胞株Skov3中的表達(dá)變化及其細(xì)胞生長變化,探討Hedgehog信號通路中Gli1和Foxm1在卵巢癌細(xì)胞株Skov3中的表達(dá)情況及其對卵巢癌細(xì)胞Skov3增殖的影響,為臨床卵巢癌的早期診斷和治療提供新的理論基礎(chǔ)。方法:1培養(yǎng)卵巢癌Skov3細(xì)胞株。2(1)實(shí)驗(yàn)組:添加Hedgehog信號通路激動劑Shh(Human Sonic Hedgehog)多肽(200ng/ml);(2)對照組:不添加Hedgehog信號通路激動劑Shh多肽。3應(yīng)用MTT法檢測各組卵巢癌Skov3細(xì)胞在24h、48h、72h的增殖情況。4采用實(shí)時熒光定量PCR法檢測各組卵巢癌Skov3細(xì)胞中24h、48h、72h Gli1m RNA和Foxm1m RNA的表達(dá)情況。5 Western blot法檢測各組卵巢癌Skov3細(xì)胞中24h、48h、72h Gli1和Foxm1蛋白表達(dá)的情況。6用SPSS21.0統(tǒng)計(jì)軟件對所有數(shù)據(jù)進(jìn)行統(tǒng)計(jì)學(xué)分析。所有數(shù)據(jù)為正態(tài)分布,以均數(shù)±標(biāo)準(zhǔn)差((?)±s)方式表示,兩組間均數(shù)比較用t檢驗(yàn),P0.05為有統(tǒng)計(jì)學(xué)意義。結(jié)果:1 MTT法檢測各組細(xì)胞增殖情況:24、48、72小時,實(shí)驗(yàn)組較對照組細(xì)胞增殖速度明顯加快,統(tǒng)計(jì)學(xué)上有顯著差異(P0.05)。2實(shí)時熒光定量PCR法檢測24h、48h、72h兩組細(xì)胞中Gli1m RNA和Foxm1m RNA的表達(dá)情況:實(shí)驗(yàn)組Gli1m RNA和Foxm1m RNA在24h、48h、72h的表達(dá)量與對照組相比均明顯升高,統(tǒng)計(jì)學(xué)上有顯著差異(P0.05)。3 Western blot法檢測24h、48h、72h兩組細(xì)胞中Gli1和Foxm1蛋白表達(dá)情況:實(shí)驗(yàn)組與對照組相比,在24h、48h、72h Gli1和Foxm1蛋白表達(dá)均明顯升高,統(tǒng)計(jì)學(xué)上有顯著差異(P0.05)。結(jié)論:1 Hedgehog信號傳導(dǎo)通路激動劑Shh多肽可增加通路活性(Gli活性),促進(jìn)卵巢癌細(xì)胞生長。2 Hedgehog信號傳導(dǎo)通路,通過高調(diào)Foxm1的表達(dá)來促進(jìn)卵巢癌細(xì)胞增長。
[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 鹵standard deviation (鹵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.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R737.31

【參考文獻(xiàn)】

相關(guān)期刊論文 前9條

1 陳丹;陳紅;董良波;楊紅;王晶晶;;Gli1與FoxM1在子宮頸癌組織和細(xì)胞中的表達(dá)及意義[J];武漢大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2016年03期

2 王艷;李剛;佐滿珍;;FoxM1剪接異構(gòu)體在上皮性卵巢癌中的表達(dá)及臨床意義[J];河北醫(yī)藥;2016年04期

3 鄭建濤;張琳;李良慶;;Gli1及Foxm1水平對胃癌轉(zhuǎn)移狀況及治療預(yù)后的影響[J];胃腸病學(xué)和肝病學(xué)雜志;2014年12期

4 劉靜波;馬玲;;卵巢癌Hedgehog通路研究進(jìn)展[J];實(shí)用癌癥雜志;2014年04期

5 劉道永;鄧軍;郭偉;張明亮;陳勇;;Gli-1蛋白在胃癌中的表達(dá)及臨床意義[J];中華全科醫(yī)學(xué);2013年09期

6 應(yīng)倩;夏慶民;鄭榮壽;張思維;陳萬青;;中國2009年宮頸癌發(fā)病與死亡分析[J];中國腫瘤;2013年08期

7 覃利菊;;卵巢癌診斷和治療的現(xiàn)狀及進(jìn)展[J];中外醫(yī)學(xué)研究;2013年19期

8 陳國慶;姚珍薇;羅欣;;轉(zhuǎn)錄因子FOXM1在上皮性卵巢癌中的表達(dá)及臨床意義[J];生命科學(xué)研究;2011年01期

9 黃海林;胡志前;王為民;王毅;蔡清萍;王強(qiáng);;Hedgehog信號傳導(dǎo)通路在乳腺癌中表達(dá)增強(qiáng)[J];基礎(chǔ)醫(yī)學(xué)與臨床;2009年05期

相關(guān)碩士學(xué)位論文 前2條

1 王俐紅;Shh和Foxm1在上皮性卵巢腫瘤組織中的表達(dá)及臨床意義[D];河北醫(yī)科大學(xué);2015年

2 趙歡;HHIP基因在子宮內(nèi)膜癌中甲基化狀態(tài)及表達(dá)[D];河北醫(yī)科大學(xué);2011年

，

本文編號：1724575