本文選題：肺腺癌 + FBXO11��； 參考：《安徽醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:研究泛素連接酶復(fù)合體(Skp1 Cul1 F-box-protein,SCF)特異性亞基FBXO11的表達(dá)對肺腺癌A549細(xì)胞遷移的影響。方法:將培養(yǎng)的肺腺癌A549細(xì)胞株分為三組:空白組(Blank組)、陰性對照RNA組(siRNA-NC組)、FBXO11干擾組(siRNA-FBXO11組),應(yīng)用Li-pofectamine 2000將siRNA-FBXO11、siRNA-NC組分別轉(zhuǎn)染至siRNA-FBXO11組和siRNA-NC組的A549細(xì)胞中。(1)應(yīng)用試劑Trizol提取A549細(xì)胞中總的RNA,RT-PCR半定量的方法檢測FBXO11的mRNA水平;(2)免疫印跡法(Western-Blot)法檢測FBXO11和Snail的蛋白水平;(3)劃痕修復(fù)實(shí)驗檢測A549細(xì)胞的遷移能力。結(jié)果:(1)RT-PCR結(jié)果示,Blank組、siRNA-NC組和siRNA-FBXO11組之間比較有差異,且差異具有統(tǒng)計學(xué)意義(P0.05)。與Blank組、siRNA-NC組比較,siRNA-FBXO11組FBXO11的mRNA水平顯著降低(P0.05),而Blank組與siRNA-NC組比較差異無統(tǒng)計學(xué)意義(P0.05)。(2)Western-Blot結(jié)果示:FBXO11蛋白水平比較,三組之間有差異,且差異具有統(tǒng)計學(xué)意義(P0.05)。與Blank組、siRNA-NC組比較,siRNA-FBXO11組FBXO11的蛋白水平顯著降低(P0.05),而Blank組與siRNA-NC組比較差異無統(tǒng)計學(xué)意義(P0.05)。Snail蛋白水平比較,三組之間有差異,且差異有統(tǒng)計學(xué)意義(P0.05)。與Blank組、siRNA-NC組比較,siRNA-FBXO11組的snail蛋白表達(dá)水平顯著增加(P0.05),而Blank組與siRNA-NC組比較無顯著性差異(P0.05)。(3)劃痕實(shí)驗結(jié)果顯示:比較三組之間的遷移能力有差異,且差異具有統(tǒng)計學(xué)意義(P0.05)。與Blank組、siRNA-NC組比較,siRNA-FBXO11組的細(xì)胞遷移能力增強(qiáng)(P0.05)。Blank組與siRNA-NC組比較差異無統(tǒng)計學(xué)意義(P0.05)。結(jié)論:FBXO11基因的表達(dá)能抑制肺腺癌A549細(xì)胞的遷移能力,這可能與Snail在細(xì)胞內(nèi)的調(diào)控有關(guān)。
[Abstract]:Aim: to study the effect of the expression of the specific subunit FBXO11 of Skp1 Cul1 and F-box-protein Cul1 on the migration of lung adenocarcinoma A549 cells. Methods: the cultured lung adenocarcinoma A549 cell line was divided into three groups: blank group (Blank group), negative control group (RNA group), siRNA-NC group (FBXO11 interference group), siRNA-FBXO11 interference group (FBXO11 group). SiRNA-FBXO11siRNA-NC group was transfected into A549 cells of siRNA-FBXO11 group and siRNA-NC group by Li-pofectamine 2000. Trizol reagent was used to extract Trizol. The mRNA level of FBXO11 in A549 cells was detected by semi-quantitative RT-PCR. Western blotting was used to detect the protein level of FBXO11 and Snail in A549 cells. The migration ability of A549 cells was detected by scratch repair assay. Results the results of RT-PCR showed that there was a significant difference between the siRNA-FBXO11 group and the control group, and the difference was statistically significant (P 0.05). Compared with the Blank group, the mRNA level of the FBXO11 in the siRNA-FBXO11 group was significantly lower than that in the siRNA-NC group, but there was no significant difference between the Blank group and the siRNA-NC group. The results of Western-Blot showed that there was a significant difference among the three groups, and the difference was statistically significant. Compared with Blank group, FBXO11 protein level of siRNA-FBXO11 group was significantly lower than that of siRNA-NC group, but there was no significant difference between Blank group and siRNA-NC group. There was significant difference among the three groups, and the difference was statistically significant (P 0.05). Compared with the Blank group, the expression of snail protein in the siRNA-FBXO11 group was significantly higher than that in the siRNA-NC group, but there was no significant difference between the Blank group and the siRNA-NC group. The results of scratch test showed that the migration ability of the three groups was different and the difference was statistically significant (P 0.05). Compared with Blank group, siRNA-FBXO11 group could enhance cell migration ability. There was no significant difference between siRNA-NC group and siRNA-NC group (P 0.05). Conclusion the expression of FBXO11 gene can inhibit the migration of lung adenocarcinoma A549 cells, which may be related to the regulation of Snail in the cells.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R734.2

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