SATB1在鼻咽癌中的表达和意义及其与鼻咽癌化疗耐药和放疗抵抗的关系

发布时间：2018-12-10 20:33

【摘要】：目的检测富含AT序列的特异结合蛋白1（specialAT-rich sequence binding protein1，SATB1）的mRNA在高成瘤高转移鼻咽癌细胞株（SUNE-1亚株-5-8F）、只成瘤不转移鼻咽癌细胞株（SUNE-1亚株-6-10B）、正常人永生化鼻咽上皮细胞株（NP69-SV40T）及SATB1蛋白在鼻咽癌（nasopharyngeal carcinoma，NPC）组织中的表达，探讨SATB1的表达与NPC临床病理因素的关系。方法采用荧光定量RT-PCR（FQ-RT-PCR）的方法检测5-8F、6-10B及NP69-SV40T细胞中SATB1的mRNA表达水平；采用免疫组化法检测72例原发性NPC和30例鼻咽黏膜慢性炎症组织（chronic nasopharyngitis tissue，CNT）中SATB1蛋白的表达，并对检测结果与NPC的临床病理参数进行统计学分析。结果 1.成功建立实时荧光定量RT-PCR的检测方法，扩增效率高(＞90%)。SATB1mRNA在5-8F细胞中的表达较NP69-SV40T细胞中的表达稍低，两者间差异无显著性（P＞0.05），5-8F细胞的表达量为6-10B细胞的4.09倍，两者间比较差异有显著性（P＜0.05）。 2. SATB1染色阳性颗粒定位于细胞核。在NPC和CNT中SATB1蛋白的阳性表达率分别为52.8%（38/72）和13.3%（4/30），与CNT比较，，NPC中SATB1表达上调，差异有统计学意义（P＜0.05）。伴有淋巴结转移和不伴淋巴结转移的NPC组织中SATB1阳性表达率分别为64.3%（27/42）和36.7%（11/30），有远处转移和无远处转移的NPC组织的阳性表达率分别为88.9%（8/9）和47.6%（30/63），两者比较，差异均有统计学意义（P＜0.05）。NPC原发灶中SATB1高表达与患者性别、年龄、T分级和临床分期无关（P＞0.05），与N分级及M分级有关（P＜0.05）。结论 1.5-8F细胞（高成瘤高转移细胞株）中SATB1mRNA的表达水平显著高于6-10B细胞（只成瘤不转移细胞株）中SATB1mRNA的表达水平，可以说明SATB1mRNA的表达水平与鼻咽癌细胞株的侵袭转移能力呈正相关。 2.NPC组织中存在着SATB1蛋白的高表达，且表达水平显著高于CNT组织，提示SATB1在NPC的发生发展中可能起着重要作用。 3.SATB1蛋白在NPC组织中的表达水平与淋巴结转移和远处转移正相关，提示SATB1可能参与NPC的侵袭转移。目的研究顺铂（DDP）及X射线对鼻咽癌5-8F细胞SATB1mRNA表达水平的影响，并探讨其相关的意义。方法以噻唑蓝（MTT）法检测不同浓度DDP处理不同时间后5-8F细胞生存率的变化；用DDP及X射线干预5-8F细胞后，提取细胞总RNA，采用荧光定量RT-PCR方法检测SATB1mRNA表达情况。结果 MTT实验表明顺铂对鼻咽癌5-8F细胞的增殖抑制存在剂量时间效应关系。顺铂及X射线处理鼻咽癌5-8F细胞后，SATB1mRNA水平均显著增高（P＜0.05），差异有统计学意义。SATB1mRNA在鼻咽癌5-8F细胞中的表达变化与顺铂的作用浓度、作用时间及X射线之间无明显的剂量时间效应关系(P＞0.05。) 结论鼻咽癌5-8F细胞经顺铂及X射线作用后，SATB1mRNA表达发生上调，推测SATB1可能参与了鼻咽癌的化疗耐药和放疗抵抗。
[Abstract]:Objective to detect the specific binding protein 1 (specialAT-rich sequence binding protein1,SATB1) mRNA rich in AT sequence in Gao Cheng high metastatic nasopharyngeal carcinoma cell line (SUNE-1 substrain-5-8F). The expression of NP69-SV40T and SATB1 protein in nasopharyngeal carcinoma (nasopharyngeal carcinoma,NPC) tissues was detected by tumorigenesis and no metastasis of nasopharyngeal carcinoma cell line (SUNE-1 substrain -6-10B), and the expression of SATB1 protein in nasopharyngeal epithelial cell line (NP69-SV40T) and normal human immortalized nasopharyngeal epithelial cell line (NP69-SV40T). To investigate the relationship between the expression of SATB1 and clinicopathological factors of NPC. Methods fluorescence quantitative RT-PCR (FQ-RT-PCR) was used to detect the mRNA expression of SATB1 in 5-8FN6-10B and NP69-SV40T cells. Immunohistochemical method was used to detect the expression of SATB1 protein in 72 cases of primary NPC and 30 cases of nasopharyngeal mucosa chronic inflammation (chronic nasopharyngitis tissue,CNT). The results and clinicopathological parameters of NPC were analyzed statistically. Result 1. The detection method of real-time fluorescent quantitative RT-PCR was successfully established. The amplification efficiency was higher than 90%. The expression of SATB1mRNA in 5-8F cells was slightly lower than that in NP69-SV40T cells, and there was no significant difference between them (P > 0. 05). The expression of 5-8F cells was 4.09 times higher than that of 6-10B cells (P < 0.05). 2. The positive granules of SATB1 staining were located in the nucleus. The positive expression rates of SATB1 protein in NPC and CNT were 52.8% (38 / 72) and 13.3% (4 / 30), respectively. Compared with CNT, the expression of SATB1 in NPC was up-regulated (P < 0. 05). The positive rates of SATB1 expression in NPC with and without lymph node metastasis were 64.3% (27 / 42) and 36.7% (11 / 30), respectively. The positive expression rates of NPC with and without distant metastasis were 88.9% (8 / 9) and 47.6% (30 / 63), respectively. The difference was statistically significant (P < 0. 05). The high expression of SATB1 was not correlated with sex, age, T grade and clinical stage (P > 0. 05), but correlated with N grade and M grade (P < 0. 05). Conclusion 1.The expression of SATB1mRNA in 5-8F cells (Gao Cheng high metastatic cell line) was significantly higher than that in 6-10B cell line (only tumorigenic non-metastatic cell line), and the expression level of SATB1mRNA in 6-10B cell line was significantly higher than that in 6-10B cell line. The expression of SATB1mRNA was positively correlated with the ability of invasion and metastasis of nasopharyngeal carcinoma cell line. There is a high expression of SATB1 protein in 2.NPC tissues, and the expression level is significantly higher than that in CNT tissues, suggesting that SATB1 may play an important role in the occurrence and development of NPC. The expression of 3.SATB1 protein in NPC was positively correlated with lymph node metastasis and distant metastasis, suggesting that SATB1 might be involved in the invasion and metastasis of NPC. Objective to investigate the effects of cisplatin (DDP) and X-ray on the expression of SATB1mRNA in 5-8F cells of nasopharyngeal carcinoma (NPC). Methods the survival rate of 5-8F cells was determined by thiazolyl (MTT) assay after different concentrations of DDP. After 5-8F cells were treated with DDP and X-ray, total RNA, was extracted and the expression of SATB1mRNA was detected by fluorescence quantitative RT-PCR. Results MTT assay showed that cisplatin inhibited the proliferation of 5-8 F cells in a dose-time dependent manner. After treated with cisplatin and X-ray, the level of SATB1mRNA in 5-8F cells of nasopharyngeal carcinoma increased significantly (P < 0. 05), the difference was statistically significant. The expression of SATB1mRNA in 5-8F cells of nasopharyngeal carcinoma and the action concentration of cisplatin were significantly increased (P < 0. 05). There was no significant dose-time effect relationship between the time of action and X-ray (P > 0.05). Conclusion the expression of SATB1mRNA in 5-8F cells of nasopharyngeal carcinoma was up-regulated by cisplatin and X-ray, suggesting that SATB1 might be involved in chemotherapeutic resistance and radiotherapy resistance of nasopharyngeal carcinoma.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R739.63

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