本文選題：肺癌 + 含WW域的氧化還原酶基因　； 參考：《江南大學(xué)》2017年碩士論文

【摘要】：目的:探究含WW域的氧化還原酶(WWOX)基因與不同惡化程度非小細(xì)胞肺癌組織和肺癌細(xì)胞的相關(guān)性、WWOX過(guò)表達(dá)/干擾對(duì)肺癌細(xì)胞侵襲性的影響、RUNX2在WWOX過(guò)表達(dá)/干擾肺癌細(xì)胞中的變化及RUNX2介導(dǎo)WWOX抑制肺癌細(xì)胞侵襲性的機(jī)制。方法:本研究選用臨床非小細(xì)胞肺癌組織標(biāo)本和多種不同惡化程度的肺癌細(xì)胞作為研究對(duì)象。應(yīng)用脂質(zhì)體法瞬時(shí)轉(zhuǎn)染外源性WWOX和RUNX2過(guò)表達(dá)質(zhì)粒;脂質(zhì)體法介導(dǎo)小干擾RNA沉默WWOX和RUNX2表達(dá);熒光實(shí)時(shí)定量PCR檢測(cè)不同處理?xiàng)l件下的WWOX、RUNX2和MMP-9 mRNA的表達(dá)含量;免疫印跡法(western blot)檢測(cè)WWOX和RUNX2蛋白的表達(dá)含量;Transwell小室檢測(cè)肺癌細(xì)胞侵襲性和遷移性;劃痕試驗(yàn)檢測(cè)肺癌細(xì)胞遷移性的改變;免疫熒光檢測(cè)肺癌細(xì)胞相關(guān)黏附因子如E-cadherin的表達(dá)情況及形態(tài)學(xué)觀察。結(jié)果:臨床組織樣本實(shí)驗(yàn)表明,相較于肺癌旁正常組織,肺癌組織中WWOX的表達(dá)出現(xiàn)減少或缺失,且在高惡化組織(II期-III期肺癌)比低惡化組織(I期肺癌)中表達(dá)要減少。細(xì)胞實(shí)驗(yàn)表明,WWOX的表達(dá)量與肺癌細(xì)胞惡化程度成反比,在肺正常上皮細(xì)胞中表達(dá)最多。針對(duì)高侵襲性但WWOX低表達(dá)的H1299細(xì)胞株,轉(zhuǎn)染W(wǎng)WOX過(guò)表達(dá)質(zhì)粒后,其侵襲性和遷移性出現(xiàn)減弱,黏附性出現(xiàn)一定的上調(diào),而再轉(zhuǎn)染RUNX2過(guò)表達(dá)質(zhì)粒后,其侵襲性出現(xiàn)回升;針對(duì)低侵襲性但WWOX高表達(dá)的NL9980細(xì)胞株,應(yīng)用小干擾RNA沉默WWOX的表達(dá)后,其侵襲性和遷移性增強(qiáng),黏附性下降,而再應(yīng)用小干擾RNA沉默RUNX2表達(dá)后,其侵襲性出現(xiàn)回降。Western blot證實(shí)WWOX對(duì)RUNX2存在抑制作用,熒光實(shí)時(shí)定量PCR證實(shí)RUNX2下游因子MMP-9在WWOX過(guò)表達(dá)細(xì)胞中出現(xiàn)下調(diào),而在WWOX沉默表達(dá)細(xì)胞中出現(xiàn)上調(diào)。結(jié)論:以上結(jié)果表明,WWOX與肺癌組織和肺癌細(xì)胞的侵襲性密切相關(guān)。RUNX2介導(dǎo)了WWOX對(duì)肺癌細(xì)胞侵襲性的抑制作用。
[Abstract]:Objective: to investigate the correlation between WW domain redox gene and lung cancer tissues and lung cancer cells with different degrees of deterioration. The effect of overexpression / interference of WWOX on the invasiveness of lung cancer cells: RUNX2 overexpression / interference in WWOX The changes in cells and the mechanism of RUNX2 mediated WWOX inhibiting the invasion of lung cancer cells. Methods: clinical non-small cell lung cancer (NSCLC) tissues and lung cancer cells with different degrees of deterioration were selected. Exogenous WWOX and RUNX2 overexpression plasmids were transiently transfected with liposome method, small interfering RNA silenced WWOX and RUNX2 expression was mediated by liposome method, and the expression of WWOX and RUNX2 was detected by real-time fluorescence quantitative PCR. Western blots were used to detect the expression of WWOX and RUNX2 protein in lung cancer cells. The invasion and migration of lung cancer cells were detected by Transwell chamber, and the changes of migration of lung cancer cells were detected by scratch test. The expression and morphology of E-cadherin in lung cancer cells were detected by immunofluorescence. Results: the clinical tissue samples showed that the expression of WWOX in lung cancer was decreased or absent compared with the normal tissues adjacent to lung cancer, and the expression of WWOX was decreased in stage II-III lung cancer than in stage I lung cancer. Cell experiments showed that the expression of WWOX was inversely proportional to the deterioration of lung cancer cells, and was most expressed in normal lung epithelial cells. For H1299 cell line with high invasiveness but low expression of WWOX, the invasiveness and mobility of H1299 cells transfected with WWOX overexpression plasmid were weakened, adhesion was up-regulated, and the invasiveness of H1299 cell line increased after retransfection of RUNX2 overexpression plasmid. For the NL9980 cell line with low invasion but high expression of WWOX, the expression of WWOX was silenced by small interfering RNA, the invasion and migration of WWOX were enhanced, and the adhesion was decreased, while the expression of RUNX2 was silenced by small interfering RNA. The inhibitory effect of WWOX on RUNX2 was confirmed by Western blot. The down-regulation of RUNX2 downstream factor MMP-9 was found in WWOX overexpression cells and up-regulated in WWOX silent expression cells by real-time quantitative PCR. Conclusion: these results suggest that WWOX is closely related to the invasion of lung cancer tissues and lung cancer cells. RUNX2 mediates the inhibitory effect of WWOX on the invasion of lung cancer cells.
【學(xué)位授予單位】：江南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R734.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 瞿俊杰;萬(wàn)小平;;WWOX表達(dá)調(diào)控、功能及作用機(jī)制的研究進(jìn)展[J];中華臨床醫(yī)師雜志(電子版);2013年14期

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本文編號(hào)：1852058