本文選題：乳腺癌 + 三陰性乳腺癌��； 參考：《南京醫(yī)科大學(xué)》2017年博士論文

【摘要】：乳腺癌是當(dāng)今世界威脅女性健康的主要疾病之一,發(fā)病率在世界范圍內(nèi)為惡性腫瘤的第2位,并且仍呈上升趨勢。乳腺癌的病理分型對于診斷、治療、判斷預(yù)后相當(dāng)重要,是臨床上較成熟的風(fēng)險評估指標(biāo)。但近年來,隨著乳腺癌的分子生物醫(yī)學(xué)逐漸發(fā)展,乳腺癌的分子分型也逐漸發(fā)展為臨床診療的重要組成部分。其中,乳腺癌細(xì)胞表面ER、PR、HER-2受體均為陰性的一類乳腺癌稱之為三陰性乳腺癌(TNBC),該類乳腺癌具有發(fā)病年齡早、易局部復(fù)發(fā)、易遠(yuǎn)處轉(zhuǎn)移、預(yù)后差等臨床特點(diǎn),并且對于內(nèi)分泌治療及靶向治療均不敏感。因此,越來越多的研究關(guān)注于該類型乳腺癌,希望找到對其有效的治療方法。微小RNA(microRNAs)為內(nèi)源性非編碼單鏈小RNA,廣泛存在于真核細(xì)胞中,具有高度保守性,可通過作用于相應(yīng)的靶基因抑制其蛋白表達(dá)來調(diào)控細(xì)胞的生長、增殖、分化、凋亡、代謝等,與疾病發(fā)生密切相關(guān)。本研究中,通過查閱文獻(xiàn),我們得到在乳腺癌組織中高表達(dá)的miR-335,為了了解miR-335在三陰性乳腺癌細(xì)胞中的功能作用及可能的調(diào)節(jié)機(jī)制,本研究從以下兩個方面進(jìn)行實(shí)驗(yàn)和探索。第一部分miR-335在MDA-MB-231細(xì)胞株和HCC-1937細(xì)胞株中的功能研究目的 探索miR-335對三陰性乳腺癌細(xì)胞株MDA-MB-231和HCC-1937的增殖、遷移能力、侵襲能力的影響。方法 運(yùn)用MTT實(shí)驗(yàn)、單克隆形成實(shí)驗(yàn)來分析miR-335對細(xì)胞增殖能力的影響,通過劃痕實(shí)驗(yàn)分析對細(xì)胞遷移能力的影響,通過Transwell小室實(shí)驗(yàn)來分析miR-335對細(xì)胞侵襲能力的影響。結(jié)果 在MDA-MB-231和HCC-1937細(xì)胞株中,miR-335上調(diào)可抑制細(xì)胞的增殖、遷移、侵襲能力,而miR-335表達(dá)下調(diào)時結(jié)果相反。結(jié)論 在MDA-MB-231和HCC-1937細(xì)胞株中,miR-335起到抑癌基因的作用,故其可能作為治療TNBC的一個候選靶點(diǎn)。第二部分miR-335的靶基因及其機(jī)制研究目的 分析miR-335可能的靶基因,并初步推測其可能的作用機(jī)制。方法 通過miR-walk、miR-base、Targetscan等軟件尋找miR-335可能的靶基因,運(yùn)用雙熒光素酶報告系統(tǒng)實(shí)驗(yàn)進(jìn)行驗(yàn)證,然后運(yùn)用Western Blot實(shí)驗(yàn)推測miR-335的作用機(jī)理。結(jié)果雙熒光素酶報告實(shí)驗(yàn)證實(shí)ROCK1作為miR-335的靶點(diǎn)成立,其為miR-335的直接靶基因之一。Western Blot實(shí)驗(yàn)結(jié)果顯示miR-335上調(diào)后ROCK1蛋白表達(dá)減少;使用miR-335 inhibitor下調(diào)miR-335可得到相反結(jié)果,進(jìn)一步證實(shí)上述結(jié)果。結(jié)論 在TNBC中,miR-335通過靶向ROCK1來抑制乳腺癌MDA-MB-231細(xì)胞株和HCC-1937細(xì)胞株的增殖、遷移及侵襲能力。
[Abstract]:Breast cancer is one of the major diseases threatening the health of women in the world. The pathological classification of breast cancer is very important for diagnosis, treatment and prognosis. But in recent years, with the development of molecular biomedicine of breast cancer, molecular classification of breast cancer has gradually developed into an important part of clinical diagnosis and treatment. Among them, the type of breast cancer with negative ERP PR-HER-2 receptors on the surface of breast cancer cells is called tri-negative breast cancer, which is characterized by early onset age, easy local recurrence, easy distant metastasis, poor prognosis and so on. And not sensitive to endocrine therapy and targeted therapy. Therefore, more and more studies focus on this type of breast cancer, hoping to find effective treatment. MicroRNAs are endogenous, non-coding, single-stranded RNAs, widely present in eukaryotic cells and highly conserved to regulate cell growth, proliferation, differentiation, apoptosis, metabolism and so on by acting on target genes to inhibit their protein expression. Is closely related to disease. In this study, we obtained the highly expressed miR-335 in breast cancer tissues. In order to understand the function of miR-335 in triple-negative breast cancer cells and its possible regulatory mechanism, we conducted experiments and exploration in the following two aspects. The function of miR-335 in MDA-MB-231 cell line and HCC-1937 cell line objective to explore the effects of miR-335 on the proliferation, migration and invasion of three negative breast cancer cell lines MDA-MB-231 and HCC-1937. Methods the effect of miR-335 on cell proliferation was analyzed by MTT assay and monoclonal formation assay. The effect of miR-335 on cell migration was analyzed by scratch test. The effect of miR-335 on cell invasion was analyzed by Transwell chamber test. Results in MDA-MB-231 and HCC-1937 cell lines, upregulation of miR-335 inhibited the proliferation, migration and invasion of MDA-MB-231 and HCC-1937 cells, but the expression of miR-335 was down-regulated. Conclusion in MDA-MB-231 and HCC-1937 cell lines, miR-335 plays an important role in tumor suppressor gene, so it may be a candidate target for the treatment of TNBC. The second part is the target gene of miR-335 and its mechanism objective to analyze the possible target gene of miR-335 and to speculate its possible mechanism of action. Methods the possible target gene of miR-335 was found by using miR-walkmiR-base Targetscan software, and verified by double-luciferase reporting system, and then the mechanism of miR-335 was deduced by Western blot. Results the double luciferase report experiment confirmed that Rock1 was the target of miR-335. Western blot showed that the expression of ROCK1 decreased after miR-335 upregulation, and the reverse result could be obtained by using miR-335 inhibitor to down-regulate miR-335. The above results are further confirmed. Conclusion the proliferation, migration and invasion of breast cancer cell line MDA-MB-231 and HCC-1937 were inhibited by targeting ROCK1 in TNBC.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：R737.9

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本文編號：1982001