【摘要】：舌鱗狀細(xì)胞癌(TSCC)是一種常見(jiàn)的口腔惡性腫瘤,發(fā)病率偏高,在我國(guó)范圍內(nèi)居口腔癌部位發(fā)生率之首,約為39.95%。其發(fā)病機(jī)理尚未十分明確,大概與多種因素均有關(guān)聯(lián),近年來(lái),其發(fā)病率有明顯的增加,而同時(shí)發(fā)病年齡趨向于年輕化。舌鱗癌的生長(zhǎng)迅速,惡性度較高,浸潤(rùn)性較強(qiáng),極易累及四周黏膜以及舌體肌肉,致使舌體活動(dòng)受限以及多項(xiàng)功能障礙。 環(huán)氧化酶(COX)作為一種限速酶,在前列腺素(PGs)的合成過(guò)程中起重要作用。已知COX存在有2種亞型,其中,COX-2已在多項(xiàng)研究中被證實(shí),其在多種腫瘤組織中呈現(xiàn)過(guò)表達(dá)現(xiàn)象。 RNA干擾(RNAi)技術(shù)是使特定的基因表達(dá)誘導(dǎo)沉默現(xiàn)象的手段之一,大量研究均表明,這一技術(shù)在逆基因分析和基因治療中的應(yīng)用潛力巨大,同時(shí)還顯現(xiàn)出了高效性、高特異性以及高穩(wěn)定性的顯著優(yōu)勢(shì)。 本實(shí)驗(yàn)研究選用慢病毒介導(dǎo)的RNA干擾技術(shù),構(gòu)建COX-2基因穩(wěn)定沉默的舌鱗癌Tca8113細(xì)胞系,并通過(guò)western-blot方法驗(yàn)證COX-2蛋白的表達(dá)水平。實(shí)驗(yàn)分為COX-2沉默組(pLKO.1shCOX-2)、陰性對(duì)照組(scr)和未處理細(xì)胞組。采用MTT的方法來(lái)觀察舌鱗癌細(xì)胞Tca8113在沉默COX-2基因后對(duì)常用化療藥物敏感性的變化。 本研究結(jié)果表明,成功建立COX-2基因沉默的舌鱗癌細(xì)胞系,通過(guò)MTT的方法檢測(cè)三組細(xì)胞48h對(duì)不同濃度平陽(yáng)霉素和順鉑的敏感性,結(jié)果顯示不同濃度藥物對(duì)COX-2基因沉默組細(xì)胞的抑制率要高于對(duì)照組,結(jié)果具有統(tǒng)計(jì)學(xué)意義。 本研究結(jié)論：沉默COX-2基因能夠增強(qiáng)舌鱗癌細(xì)胞Tca8113對(duì)化療藥物平陽(yáng)霉素和順鉑的藥物敏感性。COX-2與舌鱗癌細(xì)胞生長(zhǎng)及臨床治療密切相關(guān)。
[Abstract]:Tongue squamous cell carcinoma (TSCC) is a common oral malignant tumor with high incidence. The pathogenesis of the disease is not very clear, probably related to many factors. In recent years, the incidence of the disease has increased significantly, while the age of onset tends to be younger. Tongue squamous cell carcinoma (SCC) is characterized by rapid growth, high malignancy and strong invasiveness. It is easy to involve the surrounding mucous membrane and tongue body muscles, resulting in limited tongue body activity and multiple functional disorders. As a rate-limiting enzyme, cyclooxygenase (COX) plays an important role in the synthesis of prostaglandin (PGs). There are two known subtypes of COX, among which COX-2 has been confirmed in many studies. RNA interference (RNAi) technique is one of the methods to induce silencing of specific gene expression. A large number of studies have shown that this technique has great potential in the application of inverse gene analysis and gene therapy, but also shows the significant advantages of high efficiency, high specificity and high stability. In this study, lentivirus-mediated RNA interference was used to construct Tca8113 cell line of tongue squamous cell carcinoma with stable silencing of COX-2 gene, and the expression level of COX-2 protein was verified by western-blot method. The experiment was divided into COX-2 silencing group (pLKO.1shCOX-2), negative control group (scr) and untreated cell group. The sensitivity of Tca8113 to common chemotherapeutic drugs after COX-2 gene silencing was observed by MTT. The results showed that the cell lines of tongue squamous cell carcinoma with COX-2 gene silencing were successfully established. The sensitivity of three groups of cells to different concentrations of pingyangmycin and cisplatin was detected by MTT method. The results showed that the inhibitory rate of different concentrations of drugs on COX-2 gene silencing group was higher than that of control group, and the results were statistically significant. Conclusion: silencing COX-2 gene can enhance the sensitivity of Tca8113 to the chemotherapeutic drugs Pingyangmycin and cisplatin. COX-2 is closely related to the growth and clinical treatment of tongue squamous carcinoma cells.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R739.86

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本文編號(hào)：2290850