本文選題：喉癌 + RKIP　； 參考：《吉林大學》2010年博士論文

【摘要】： 探討高效表達RKIP基因聯(lián)合化療藥物對人喉鱗癌細胞的作用,為喉癌基因聯(lián)合化療減低喉癌化療抵抗提供了理論依據(jù)。應用DNA克隆技術構建干涉和高效表達RKIP的重組質(zhì)粒,并進行測序鑒定和效果鑒定;應用真核細胞轉染技術將重組質(zhì)粒轉染人喉癌Hep-2細胞,觀察RKIP表達水平變化對喉鱗癌細胞增殖、周期、凋亡及侵襲的影響,并分析其作用機制;高效表達RKIP的重組質(zhì)粒聯(lián)合順鉑后,探討其減低喉癌化療抵抗和增強喉癌對順鉑敏感性的可能性。(1)經(jīng)酶切和測序證實,成功構建重組質(zhì)粒。熒光定量RT-PCR和Western印跡檢測RKIP mRNA和蛋白的表達,高效表達組升高,干涉組表達降低,其中干涉組的RKIP-shRNA501抑制作用最為明顯。(2)MTT檢測結果顯示,高效表達組出現(xiàn)增殖抑制,干涉組出現(xiàn)增殖促進。FCM檢測結果顯示,高效表達組的G1期細胞增多,出現(xiàn)明顯的凋亡峰。干涉組的G1期細胞減少,未出現(xiàn)明顯的凋亡峰。Matrigel侵襲實驗結果顯示,高效表達組細胞侵襲力減弱,干涉組細胞侵襲力增強。MAPK通路的改變或許是RKIP引起喉鱗癌Hep-2細胞增殖、周期、凋亡、侵襲等改變的原因或部分原因。(3)高效表達RKIP的重組質(zhì)粒聯(lián)合順鉑后,MTT檢測結果顯示,細胞增殖受到明顯抑制。FCM檢測結果顯示,細胞凋亡明顯增強,MAPK通路的改變或許是RKIP-pIRES2-EGFP增強DDP對喉鱗癌Hep-2細胞影響的原因或部分原因。高效表達RKIP基因聯(lián)合化療對人喉鱗癌具有顯著的治療效果。
[Abstract]:To investigate the effect of high expression of RKIP gene combined with chemotherapeutic drugs on human laryngeal squamous carcinoma cells, and to provide a theoretical basis for the combination of laryngeal oncogene and chemotherapy to reduce the chemotherapeutic resistance of laryngeal carcinoma.The recombinant plasmids expressing RKIP were constructed by DNA cloning, sequenced and identified, and the recombinant plasmids were transfected into human laryngeal carcinoma Hep-2 cells by eukaryotic cell transfection.To observe the effect of RKIP expression on proliferation, cell cycle, apoptosis and invasion of laryngeal squamous cell carcinoma cells, and to analyze its mechanism.To investigate the possibility of reducing chemotherapeutic resistance and enhancing the sensitivity of laryngeal carcinoma to cisplatin, the recombinant plasmid was successfully constructed by restriction endonuclease digestion and sequencing.Fluorescence quantitative RT-PCR and Western blotting were used to detect the expression of RKIP mRNA and protein. The expression of RKIP mRNA and protein increased in high expression group and decreased in intervention group. The inhibitory effect of RKIP-shRNA501 in intervention group was the most obvious. The results showed that proliferation inhibition appeared in high expression group.The results of proliferation-promoting. FCM assay showed that the G1 phase cells in the high expression group increased and the apoptotic peak appeared.In the intervention group, the G1 phase of cells decreased, and no obvious apoptotic peak. Matrigel invasion assay showed that the cell invasion of the high expression group was weakened, and the increase of the cell invasion and MAPK pathway in the intervention group might be due to the proliferation and cycle of Hep-2 cells induced by RKIP.The reason or part of apoptosis, invasion, etc.) the recombinant plasmid expressing RKIP efficiently combined with cisplatin showed that the cell proliferation was obviously inhibited.The effect of RKIP-pIRES2-EGFP on DDP in laryngeal squamous cell carcinoma (LSCC) might be due to the change of apoptosis.High expression of RKIP gene combined with chemotherapy has a significant therapeutic effect on human laryngeal squamous cell carcinoma.
【學位授予單位】：吉林大學
【學位級別】：博士
【學位授予年份】：2010
【分類號】：R739.65

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