【摘要】：[目的]胃癌是最常見的消化道惡性腫瘤,最近幾年發(fā)病率逐年升高。超過半數(shù)患者初次就診時(shí)疾病已進(jìn)展至中晚期,即使在通過手術(shù)根治+輔助性放化療,治療效果仍不理想。胃癌轉(zhuǎn)移是腫瘤最重要的惡性特征,也是導(dǎo)致腫瘤患者死亡的最主要原因。腫瘤的轉(zhuǎn)移是一個(gè)多步驟的過程,在腫瘤轉(zhuǎn)移初期過程中腫瘤細(xì)胞發(fā)生上皮間質(zhì)轉(zhuǎn)化(Epithelial-Mesenchymal Transition,EMT)。目前,研究發(fā)現(xiàn),轉(zhuǎn)化生長因子與Wnt/β-catenin信號(hào)通路在腫瘤EMT進(jìn)程中發(fā)揮著重要作用。我們的目的是探討胃癌轉(zhuǎn)移過程中發(fā)生EMT后的表型改變以及Wnt信號(hào)通路在其中的作用機(jī)制,為胃癌轉(zhuǎn)移提供新的理論依據(jù),為胃癌轉(zhuǎn)移的靶向治療提供實(shí)驗(yàn)依據(jù)。[方法]將人胃癌細(xì)胞株MKN28的細(xì)胞接種于培養(yǎng)基中,待細(xì)胞長至對(duì)數(shù)期80%匯合度時(shí),進(jìn)行傳代。隨后加入不同濃度TGF-β 1(5/10/20/50ng/ml),做72h誘導(dǎo)處理后,通過CCK8、細(xì)胞周期試驗(yàn)、侵襲實(shí)驗(yàn)及在倒置顯微鏡下實(shí)時(shí)觀察細(xì)胞表型變化并與親代MKN28細(xì)胞株作對(duì)比。并且通過Western Blot方法檢測MET標(biāo)志物β-catenin,以及Wnt/β-catenin信號(hào)通路相關(guān)基因Wnt3a、β-catenin、CyclinD1的表達(dá)與原腫瘤細(xì)胞做對(duì)比。[結(jié)果]實(shí)驗(yàn)發(fā)現(xiàn)不同濃度TGFβ 1在體外能夠誘導(dǎo)胃癌細(xì)胞發(fā)生上皮間質(zhì)轉(zhuǎn)化。使得胃癌細(xì)胞出現(xiàn)細(xì)胞形態(tài)改變、細(xì)胞之間黏附力減弱、細(xì)胞活力增加,增殖、侵襲能力增強(qiáng)。與此同時(shí)還可激活胃癌細(xì)胞中的Wnt3a信號(hào)通路,上調(diào)Wnt3a、β-catenin、CyclinD1的表達(dá);但實(shí)驗(yàn)結(jié)果與TGFβ1的濃度未出現(xiàn)相關(guān)性,在該試驗(yàn)中存在最佳誘導(dǎo)濃度為1Ong/ml。[結(jié)論]1.胃癌細(xì)胞在不同濃度TGFβ1的誘導(dǎo)下可發(fā)生EMT轉(zhuǎn)化,其細(xì)胞形態(tài)改變、細(xì)胞之間黏附力減弱、細(xì)胞活力增加,增殖、侵襲能力增強(qiáng)。2.TGFβ 1可以有效激活Wnt3a信號(hào)通路促進(jìn)胃癌細(xì)胞進(jìn)一步發(fā)生EMT。3.TGFβ 1在誘導(dǎo)過程中胃癌細(xì)胞表達(dá)產(chǎn)物的變化與其濃度未出現(xiàn)相關(guān)性。
[Abstract]:Objective: gastric cancer is the most common malignant tumor of digestive tract. More than half of the patients had advanced to advanced stage at the time of first visit, and the effect of treatment was still not satisfactory even after radical operation with adjuvant radiotherapy and chemotherapy. Metastasis of gastric cancer is the most important malignant feature and the main cause of death in cancer patients. Tumor metastasis is a multistep process in which tumor cells undergo epithelial interstitial transformation (Epithelial-Mesenchymal Transition,EMT) in the early stages of tumor metastasis. At present, it has been found that transforming growth factor and Wnt/ 尾-catenin signaling pathway play an important role in tumor EMT process. Our aim is to investigate the phenotypic changes after EMT and the mechanism of Wnt signaling pathway in gastric cancer metastasis, to provide a new theoretical basis for gastric cancer metastasis, and to provide experimental evidence for the targeted treatment of gastric cancer metastasis. [methods] the cells of human gastric cancer cell line MKN28 were inoculated into culture medium and subcultured when the cells reached the logarithmic phase of 80% confluence. Then different concentrations of TGF- 尾 1 (5/10/20/50ng/ml) were added. After 72 hours of induction, the phenotypic changes of the cells were observed by CCK8, cell cycle test, invasion test and real-time observation under inverted microscope. The results were compared with those of the parent MKN28 cell line. The expression of MET marker 尾-catenin, and Wnt/ 尾-catenin signaling pathway related gene Wnt3a, 尾-catenin,CyclinD1 was detected by Western Blot method. [results] it was found that different concentrations of TGF 尾 1 could induce epithelial mesenchymal transformation of gastric cancer cells in vitro. The cell morphology of gastric cancer cells was changed, the adhesion between cells was weakened, the cell viability was increased, proliferation and invasion were enhanced. At the same time, the Wnt3a signaling pathway was activated and the expression of Wnt3a, 尾-catenin,CyclinD1 was up-regulated in gastric cancer cells, but there was no correlation between the results and the concentration of TGF 尾 _ 1. The optimal induction concentration was 1 Ong / ml. [conclusion] 1. Under the induction of different concentrations of TGF 尾 1, gastric cancer cells can undergo EMT transformation. The cell morphology changes, the adhesion between cells decreases, the cell viability increases and the cell proliferates. 2.TGF 尾 1 could effectively activate the Wnt3a signaling pathway to promote the further development of EMT.3.TGF 尾 1 in gastric cancer cells. There was no correlation between the expression of EMT.3.TGF 尾 1 and the concentration of EMT.3.TGF 尾 1 in gastric cancer cells.
【學(xué)位授予單位】：昆明醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 John C.Layke;Peter P.Lopez;本刊編輯部;;胃癌:診斷和治療方案的選擇[J];中國全科醫(yī)學(xué);2015年03期

2 曹中正;丁連安;牛冬光;權(quán)琳;;胃癌外周血循環(huán)腫瘤細(xì)胞水平及其臨床意義[J];臨床腫瘤學(xué)雜志;2014年06期

3 鄒文斌;李兆申;;中國胃癌發(fā)病率及死亡率研究進(jìn)展[J];中國實(shí)用內(nèi)科雜志;2014年04期

4 曲福杰;潘立明;孫抒;;CyclinD1和CD15蛋白在結(jié)直腸癌中的表達(dá)及其臨床意義[J];延邊大學(xué)醫(yī)學(xué)學(xué)報(bào);2014年01期

5 Bin Zhang;Lin-Hua Ji;Wei Liu;Gang Zhao;Zhi-Yong Wu;;Skp2-RNAi suppresses proliferation and migration of gallbladder carcinoma cells by enhancing p27 expression[J];World Journal of Gastroenterology;2013年30期

6 李志章;李欽;;TGF-β/Smads信號(hào)通路及其在腫瘤中的作用[J];醫(yī)學(xué)綜述;2011年06期

7 余鋒;張妤;施樂華;殷正豐;;循環(huán)腫瘤細(xì)胞檢測的臨床應(yīng)用[J];世界華人消化雜志;2010年22期

8 周晴接;楊建民;;循環(huán)腫瘤細(xì)胞研究進(jìn)展[J];世界華人消化雜志;2010年11期

9 田冬艷;王德華;劉永紅;齊廣濤;邸麗雙;;Survivin和CyclinD1在卵巢癌中的表達(dá)和相關(guān)性[J];天津醫(yī)科大學(xué)學(xué)報(bào);2009年01期

10 宮奇林;林梅青;時(shí)昌文;孫濤;;Twist基因檢測對(duì)大腸癌臨床病理診斷價(jià)值的探討[J];中華腫瘤防治雜志;2008年16期

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