發(fā)布時(shí)間：2018-05-08 22:42

  本文選題：卵巢癌 + MAP3K3��； 參考：《石河子大學(xué)》2017年碩士論文

【摘要】：目的:干擾MAP3K3表達(dá)和/或阻斷NF-κB信號通路,檢測MAP3K3在卵巢癌細(xì)胞中的表達(dá),結(jié)合NF-κB信號通路相關(guān)因子及凋亡相關(guān)蛋白的表達(dá)變化,分析和探討MAP3K3介導(dǎo)NF-κB信號通路在卵巢癌化療抵抗中的作用。方法:分別用特異性干擾MAP3K3表達(dá)質(zhì)粒和MAP3K3真核表達(dá)質(zhì)粒轉(zhuǎn)染內(nèi)源性高表達(dá)和/或低表達(dá)MAP3K3的卵巢癌細(xì)胞系SKOV3和OV2008,采用Western Blot和qRT-PCR檢測轉(zhuǎn)染效率;運(yùn)用MTT方法檢測干擾前后卵巢癌細(xì)胞對順鉑、紫杉醇和TNF-α敏感性的變化,TUNEL法檢測細(xì)胞凋亡;通過使用NF-κB信號通路特異性阻斷劑QNZ,觀察順鉑處理后NF-κB信號通路關(guān)鍵因子和凋亡相關(guān)因子表達(dá)的變化情況。結(jié)果:(1)MAP3K3蛋白和mRNA在SKOV3細(xì)胞中內(nèi)源性高表達(dá),在OV2008細(xì)胞中低表達(dá),OV2008細(xì)胞對順鉑的敏感性顯著高于SKOV3細(xì)胞;(2)在SKOV3中敲低MAP3K3表達(dá)后,順鉑誘導(dǎo)細(xì)胞凋亡的數(shù)目顯著增加,細(xì)胞對順鉑敏感性增強(qiáng),生長速度減慢,同時(shí)NF-κB信號通路相關(guān)因子p-P65、p-IκBα和抗凋亡蛋白Bcl-2表達(dá)下調(diào),促凋亡蛋白cleaved-caspase3、BAX表達(dá)上調(diào);(3)在OV2008中過表達(dá)MAP3K3后,順鉑誘導(dǎo)細(xì)胞凋亡的數(shù)目顯著減少,細(xì)胞對順鉑的敏感性減弱,生長加快,同時(shí)NF-κB信號通路關(guān)鍵因子p-P65、p-IκBα和抗凋亡蛋白Bcl-2表達(dá)上調(diào),促凋亡蛋白cleaved-caspase3、BAX表達(dá)下調(diào);(4)在SKOV3中抑制MAP3K3表達(dá)后細(xì)胞對紫杉醇和TNF-α的敏感性增強(qiáng);在OV2008中過表達(dá)MAP3K3后細(xì)胞對紫杉醇和TNF-α的敏感性減弱;阻斷NF-κB信號通路,兩株細(xì)胞對紫杉醇和TNF-α的敏感性增強(qiáng)。結(jié)論:MAP3K3在卵巢癌細(xì)胞系中表達(dá)水平的差異可能與卵巢癌細(xì)胞的化療抵抗相關(guān)。MAP3K3高表達(dá)促進(jìn)了NF-κB信號通路激活以及下游抗凋亡蛋白的表達(dá),抑制促凋亡蛋白表達(dá),降低了化療藥物對卵巢癌細(xì)胞的誘導(dǎo)凋亡能力,提示MAP3K3可能是通過介導(dǎo)NF-κB信號通路促進(jìn)卵巢癌細(xì)胞發(fā)生化療抵抗。
[Abstract]:Aim: to interfere with the expression of MAP3K3 and / or block NF- 魏 B signaling pathway, detect the expression of MAP3K3 in ovarian cancer cells, and combine the changes of NF- 魏 B signal pathway related factors and apoptosis-related proteins. To investigate the role of NF- 魏 B signaling pathway mediated by MAP3K3 in chemotherapeutic resistance of ovarian cancer. Methods: ovarian cancer cell lines SKOV3 and OV2008 were transfected with specific interfering MAP3K3 expression plasmids and MAP3K3 eukaryotic expression plasmids. The transfection efficiency was detected by Western Blot and qRT-PCR. The sensitivity of ovarian cancer cells to cisplatin, paclitaxel and TNF- 偽 was detected by MTT method. Apoptosis was detected by Tunel method. The expression of key factors and apoptosis-related factors in NF- 魏 B signaling pathway after cisplatin treatment was observed by using QNZ, a specific inhibitor of NF- 魏 B signaling pathway. Results the endogenous expression of MAP3K3 protein and mRNA in SKOV3 cells was significantly higher than that in OV2008 cells. The sensitivity to cisplatin in OV2008 cells was significantly higher than that in SKOV3 cells (P < 0.05). The number of apoptosis induced by cisplatin increased significantly when MAP3K3 expression was low in SKOV3. At the same time, the expression of NF- 魏 B signaling pathway related factor p-P65, p-I 魏 B 偽 and anti-apoptotic protein Bcl-2 was down-regulated, and the expression of cleaved-caspase3mBAX was up-regulated in OV2008. The number of apoptosis induced by cisplatin was significantly reduced, the sensitivity of cells to cisplatin was weakened, and the growth rate was accelerated. Meanwhile, the expression of p-P65 p-I 魏 B 偽 and anti-apoptotic protein Bcl-2 was up-regulated in NF- 魏 B signaling pathway. After inhibiting the expression of MAP3K3 in SKOV3, the sensitivity of cells to paclitaxel and TNF- 偽 was enhanced, the sensitivity of cells to paclitaxel and TNF- 偽 was weakened after overexpression of MAP3K3 in OV2008, and the signal pathway of NF- 魏 B was blocked. The sensitivity of the two cells to paclitaxel and TNF- 偽 was enhanced. Conclusion the difference of the expression level of protein MAP3K3 in ovarian cancer cell line may be related to the chemotherapeutic resistance of ovarian cancer cell line. The overexpression of MAP3K3 promotes the activation of NF- 魏 B signaling pathway and the expression of downstream anti-apoptotic protein, and inhibits the expression of pro-apoptotic protein. The ability of apoptosis induced by chemotherapeutic drugs was reduced, suggesting that MAP3K3 may promote chemotherapeutic resistance by mediating NF- 魏 B signaling pathway.
【學(xué)位授予單位】：石河子大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R737.31

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