慢病毒载体沉默COX-2基因对舌鳞癌Tca8113细胞株的药物敏感性的影响
发布时间：2018-10-24 08:23
【摘要】：舌鳞状细胞癌(TSCC)是一种常见的口腔恶性肿瘤,发病率偏高,在我国范围内居口腔癌部位发生率之首,约为39.95%。其发病机理尚未十分明确,大概与多种因素均有关联,近年来,其发病率有明显的增加,而同时发病年龄趋向于年轻化。舌鳞癌的生长迅速,恶性度较高,浸润性较强,极易累及四周黏膜以及舌体肌肉,致使舌体活动受限以及多项功能障碍。 环氧化酶(COX)作为一种限速酶,在前列腺素(PGs)的合成过程中起重要作用。已知COX存在有2种亚型,其中,COX-2已在多项研究中被证实,其在多种肿瘤组织中呈现过表达现象。 RNA干扰(RNAi)技术是使特定的基因表达诱导沉默现象的手段之一,大量研究均表明,这一技术在逆基因分析和基因治疗中的应用潜力巨大,同时还显现出了高效性、高特异性以及高稳定性的显著优势。 本实验研究选用慢病毒介导的RNA干扰技术,构建COX-2基因稳定沉默的舌鳞癌Tca8113细胞系,并通过western-blot方法验证COX-2蛋白的表达水平。实验分为COX-2沉默组(pLKO.1shCOX-2)、阴性对照组(scr)和未处理细胞组。采用MTT的方法来观察舌鳞癌细胞Tca8113在沉默COX-2基因后对常用化疗药物敏感性的变化。 本研究结果表明,成功建立COX-2基因沉默的舌鳞癌细胞系,通过MTT的方法检测三组细胞48h对不同浓度平阳霉素和顺铂的敏感性,结果显示不同浓度药物对COX-2基因沉默组细胞的抑制率要高于对照组,结果具有统计学意义。 本研究结论：沉默COX-2基因能够增强舌鳞癌细胞Tca8113对化疗药物平阳霉素和顺铂的药物敏感性。COX-2与舌鳞癌细胞生长及临床治疗密切相关。
[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.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.86
本文编号：2290850
[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.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.86
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相关期刊论文 前2条
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，本文编号：2290850
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