發(fā)布時(shí)間：2018-03-16 20:04

  本文選題：子宮內(nèi)膜癌　切入點(diǎn)：miR-124　出處：《南昌大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：研究目的：探究miR-124在子宮內(nèi)膜癌中的表達(dá)及其對(duì)子宮內(nèi)膜癌細(xì)胞惡性生物學(xué)表型的影響及其機(jī)制，為今后的內(nèi)膜癌早期診斷及生物治療提供實(shí)驗(yàn)依據(jù)。方法：我們首先利用stem-loop RT-PCR法檢測(cè)miR-124在35對(duì)子宮內(nèi)膜癌組織及其對(duì)應(yīng)的瘤旁正常組織的表達(dá)；分別運(yùn)用CCK8法、流式細(xì)胞技術(shù)、細(xì)胞劃痕實(shí)驗(yàn)以及transwell實(shí)驗(yàn)，檢測(cè)細(xì)胞增殖、周期、凋亡、遷移及侵襲的變化；通過(guò)進(jìn)一步的生物信息學(xué)靶基因的預(yù)測(cè)miR-124的靶基因，并運(yùn)用雙螢光報(bào)告以及Western Blotting實(shí)驗(yàn)驗(yàn)證；進(jìn)一步運(yùn)用Rescue實(shí)驗(yàn)驗(yàn)證該靶基因在miR-124介導(dǎo)的抑癌功能中的作用。 結(jié)果：miR-124在子宮內(nèi)膜癌組織中的表達(dá)呈明顯下調(diào)趨勢(shì),提示miR-124可能在子宮內(nèi)膜癌發(fā)生發(fā)展中發(fā)揮抑癌基因的功能；恢復(fù)子宮內(nèi)膜癌來(lái)源的HEC-1B細(xì)胞中miR-124表達(dá)能夠明顯抑制細(xì)胞的增殖、侵襲和遷移，抑制細(xì)胞周期進(jìn)程，誘導(dǎo)細(xì)胞早期凋亡的發(fā)生；通過(guò)進(jìn)一步的生物信息學(xué)靶基因的預(yù)測(cè)提示信號(hào)轉(zhuǎn)導(dǎo)與轉(zhuǎn)錄激活因子(signal transducer and transcription activator3, STAT3)是miR-124的靶基因之一。雙螢光報(bào)告以及Western Blotting實(shí)驗(yàn)證實(shí)，miR-124能夠明顯抑制HEC-1B細(xì)胞內(nèi)STAT3基因的轉(zhuǎn)錄，，轉(zhuǎn)染后HEC-1B細(xì)胞內(nèi)STAT3的mRNA及蛋白水平均得到明顯抑制；進(jìn)一步敲低HEC-1B細(xì)胞中STAT3的表達(dá)與過(guò)表達(dá)miR-124結(jié)果一致，能夠明顯抑制HEC-1B細(xì)胞的惡性生物學(xué)表型，提示miR-124介導(dǎo)的抑癌功能可能是部分依賴于靶向調(diào)控STAT3通路產(chǎn)生的；同時(shí)運(yùn)用IL-6處理后能夠部分性的恢復(fù)細(xì)胞中STAT3的表達(dá)，以及減弱miR-124對(duì)HEC-1B細(xì)胞惡性表型的抑制作用，這一結(jié)果表明在子宮內(nèi)膜癌細(xì)胞中，miR-124介導(dǎo)的抑癌功能是直接通過(guò)靶向STAT3產(chǎn)生的； 結(jié)論：我們的結(jié)果進(jìn)一步確定了miR-124在EC發(fā)生發(fā)展中的抑癌基因功能及其機(jī)制，同時(shí)為miR-124用于EC的診斷和治療奠定了生物學(xué)基礎(chǔ)。
[Abstract]:Objective: to investigate the expression of miR-124 in endometrial carcinoma and its effect on the malignant biological phenotype of endometrial cancer cells and its mechanism. Methods: stem-loop RT-PCR method was used to detect the expression of miR-124 in 35 pairs of endometrial carcinoma and its adjacent normal tissues, respectively, using CCK8 method and flow cytometry. Cell scratch assay and transwell assay were used to detect the changes of cell proliferation, cell cycle, apoptosis, migration and invasion. The target genes of miR-124 were predicted by further bioinformatics, and verified by double fluorescence report and Western Blotting experiment. Rescue assay was used to verify the role of the target gene in the tumor suppressor function mediated by miR-124. Results the expression of 10 miR-124 was down-regulated in endometrial carcinoma, suggesting that miR-124 might play a role as a tumor suppressor gene in the carcinogenesis and development of endometrial carcinoma. The expression of miR-124 in HEC-1B cells derived from endometrial carcinoma can significantly inhibit cell proliferation, invasion and migration, inhibit the progress of cell cycle and induce early apoptosis. The prediction of further bioinformatics target genes suggested that signal transducer and transcription activator 3 (STAT3) was one of the target genes of miR-124. Double fluorescence reports and Western Blotting experiments showed that the signal transduction and transcription activator 3 (STAT3) could significantly inhibit the transcription of STAT3 gene in HEC-1B cells. After transfection, the mRNA and protein levels of STAT3 in HEC-1B cells were significantly inhibited, and the expression of STAT3 in HEC-1B cells was consistent with the results of overexpression of miR-124, which could inhibit the malignant biological phenotype of HEC-1B cells. The results suggest that the inhibition function of miR-124 may be partly dependent on the targeting regulation of STAT3 pathway, at the same time, it can partially restore the expression of STAT3 in the cells treated with IL-6, and attenuate the inhibitory effect of miR-124 on the malignant phenotype of HEC-1B cells. These results suggest that the inhibitory effect of miR-124 is directly mediated by targeted STAT3 in endometrial cancer cells. Conclusion: our results further confirm the function and mechanism of tumor suppressor gene of miR-124 in the development of EC and lay a biological foundation for the diagnosis and treatment of EC by miR-124.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R737.33

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 郎景和;子宮內(nèi)膜癌診治的幾個(gè)問(wèn)題[J];中華婦產(chǎn)科雜志;2000年05期

2 ;子宮內(nèi)膜癌發(fā)病率躍居?jì)D科惡性腫瘤首位[J];中華婦幼臨床醫(yī)學(xué)雜志(電子版);2011年06期

本文編號(hào)：1621449