分化型甲状腺癌BRAF基因突变、TERT启动子突变与PPARγ、NIS、TSHR表达的相关性

发布时间：2018-02-13 06:53

本文关键词： 甲状腺癌基因突变过氧化物酶增殖物激活受体-γ 钠碘转运体促甲状腺激素受体　出处：《兰州大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:探讨分化型甲状腺癌(DTC)鼠类肉瘤滤过性毒菌致癌同源体B1(BRAF)基因突变、端粒酶逆转录酶(TERT)启动子突变和过氧化物酶增殖物激活受体γ(PPARγ)、钠碘转运体(NIS)及促甲状腺激素受体(TSHR)表达及临床意义,并研究BRAF基因突变、TERT启动子突变对PPARγ、NIS及TSHR表达的影响及相关性。方法:收集病理确诊的DTC患者229例,并取同期手术治疗的结节性甲状腺肿52例及正常甲状腺组织31例。提取DNA采用PCR扩增产物直接测序法检测BRAF基因V600E点突变、TERT启动子C228T及C250T点突变,免疫组织化学SP法检测PPARγ、NIS及TSHR蛋白表达。采用SPSS 22.0统计软件Pearson chi-square(χ2)分析和非参数Spearman相关分析,P0.05为差异有统计学意义。结果:1.结节性甲状腺肿及正常甲状腺组织中均未发现BRAF V600E基因突变和TERT启动子C228T及C250T点突变。2.DTC患者BRAF V600E基因突变率为62.0%,与性别、年龄、肿瘤大小、淋巴结转移及复发危险度分层均无关(P0.05)。3.DTC患者TERT启动子突变率为7.9%,其中C250T位点突变率为7.0%,C228T位点为0.9%,与性别、年龄及复发危险度分层相关(P0.05)。4.DTC患者TERT启动子突变同时伴BRAF V600E基因突变率为61.1%,与淋巴结转移及复发危险度分层相关(P0.05)。5.DTC患者BRAF V600E突变组PPARγ阳性率为59.9%,高于BRAF V600E野生组26.4%(P0.05);DTC患者PPARγ阳性率在只存在BRAF V600E突变组(BRAF+、TERT-)和BRAF V600E、TERT启动子同时突变组(BRAF+、TERT+)分别为59.5%和63.6%,均高于BRAF V600E、TERT启动子均未突变组(BRAF-、TERT-)25.0%(P0.05)。6.NIS在DTC中阳性率为55.5%,低于结节性甲状腺肿78.9%及正常甲状腺组织77.4%(P0.05);DTC患者BRAF+、TERT+组NIS阳性率18.2%,低于BRAF-、TERT-组51.3%(P0.05);DTC患者NIS细胞浆表达者48.2%发生于BRAF V600E突变组高于BRAF V600E野生组29.5%;细胞膜表达者70.5%发生于BRAF V600E野生组高于BRAF V600E突变组51.8%(P0.05)。7.TSHR在DTC中阳性率为41.0%,与肿瘤大小有关(P0.05);DTC患者BRAF V600E突变组TSHR阳性率48.6%高于BRAF V600E野生组28.7%(P0.05);DTC患者TSHR阳性率在BRAF+、TERT-组,BRAF-、TERT+组及BRAF+、TERT+组分别为46.6%、85.7%、72.7%,均高于BRAF-、TERT-组23.8%(P0.05)。结论:1.检测BRAF V600E基因突变和TERT启动子C228T及C250T点突变有助于鉴别甲状腺肿瘤良、恶性。2.DTC患者BRAF V600E基因突变率较高,但单独检测BRAF V600E突变尚难对DTC预后评估有指导。TERT启动子在DTC中突变率较低,且以C250T位点突变更多见。联合检测BRAF V600E和TERT启动子突变对DTC预后评估具有指导意义。3.DTC患者PPARγ表达受BRAF V600E和TERT启动子突变共同影响,提示BRAF V600E和TERT启动子突变可能与PPARγ共同作用促进肿瘤组织增生,支持PPARγ作为促癌因子的观点。4.DTC患者NIS表达定位与BRAF V600E有关,NIS表达降低与BRAF V600E和TERT启动子同时突变有关,检测BRAF V600E和TERT启动子同时突变可间接预测DTC组织的摄碘能力。5.DTC患者TSHR表达受BRAF V600E和TERT启动子突变共同影响。
[Abstract]:Objective: to investigate the mutation of the oncogenic homologue B1BRAF of murine sarcomas derived from differentiated thyroid carcinoma (DTC). Expression and clinical significance of telomerase reverse transcriptase promoter mutation and peroxidase proliferator activated receptor 纬 PPAR 纬, sodium iodide transporter (NIS) and thyrotropin receptor (TSH). To study the effect and correlation of BRAF gene mutation and TERT promoter mutation on the expression of PPAR 纬 -NIS and TSHR. Methods: 229 patients with pathologically diagnosed DTC were collected. DNA was extracted from 52 cases of nodular goiter and 31 cases of normal thyroid tissue in the same period of operation. PCR amplification products were used to detect C228T and C250T point mutations in the promoter of BRAF gene V600E site-mutation, C228T and C250T, respectively. Immunohistochemical SP method was used to detect the expression of PPAR 纬 nis and TSHR protein. There was significant difference between SPSS 22.0 statistical software Pearson chi-square (蠂 2) and nonparametric Spearman correlation analysis (P0.05). Results: 1. Nodular goiter and normal thyroid tissue were all different. No mutation of BRAF V600E gene and point mutation of C228T and C250T of TERT promoter. 2. The mutation rate of BRAF V600E gene in patients with DTC was 62.0%, which was related to sex. Age, tumor size, lymph node metastasis and recurrence risk stratification were not correlated with the mutation rate of TERT promoter in DTC patients. The mutation rate of C250T locus was 7.0T and C228T locus was 0.9g with sex. The mutation rate of TERT promoter with BRAF V600E gene was 61.1in patients with age and recurrence risk stratification. The positive rate of PPAR 纬 in BRAF V600E mutation group was 59.9in patients with BRAF V600E mutation, which was higher than that in wild BRAF V600E group, which was associated with lymph node metastasis and recurrence risk stratification (P 0.05). 5. The positive rate of PPAR 纬 in BRAF V600E mutation group was higher than that in BRAF V600E wild group. The positive rate of PPAR 纬 in BRAF V600E mutation group and BRAF V600Etert promoter mutation group was 59.5% and 63.6, respectively, which was higher than that in BRAF V600 Etert promoter non-mutation group, BRAF-TERT promoter with no mutation, BRAF-TERT positive rate in DTC was 55.5, which was lower than that in nodular goiter 78.9%. The positive rate of NIS in the BRAF group was 18.2and was lower than that in the BRAF-TERT- group, which was lower than that in the BRAF V600E mutation group and the BRAF V600E wild group. The expression rate of NIS in the BRAF V600E mutation group was higher than that in the BRAF V600E wild group, and the cell membrane expression rate in the BRAF V600E wild group was higher than that in the BRAF V600E process. The positive rate of TSHR in the BRAF V600E mutation group was 48.6% higher than that in the BRAF V600E mutant group, and the TSHR positive rate in BRAF TERT- group BRAF-TERT group and BRAF tert group was 46.60.75.72.70.The positive rate of TSHR was higher than that of BRAF-T- group 23.8TERP0.052.72.70.The positive rate of TSHR in BRAF TERT- group was higher than that in BRAF V600E mutant group (28.75.72.72.72.77.The positive rate of TSHR in BRAF TERT- group BRAF-TERT group and BRAF tert group was 46.65.72.72.77.The positive rate of TSHR in BRAF V600E mutation group was higher than that in BRAF V600E wild group 28.75.72.72.71.Conclusion:. The mutation of BRAF V600E gene and the point mutation of C228T and C250T promoter of TERT are helpful in differentiating benign thyroid tumors. 2.The mutation rate of BRAF V600E gene in DTC patients was high, but it was difficult to evaluate the prognosis of DTC by detecting BRAF V600E mutation alone. The mutation rate of TERT promoter was lower in DTC. The combined detection of BRAF V600E and TERT promoter mutations may be helpful in evaluating the prognosis of DTC. 3. The expression of PPAR 纬 in DTC patients is affected by BRAF V600E and TERT promoter mutations. The results suggest that the mutation of BRAF V600E and TERT promoter may promote the proliferation of tumor tissue together with PPAR 纬. The view that PPAR 纬 is used as a carcinogenic factor. 4. The location of NIS expression in DTC patients is related to the decrease of BRAF V600E expression and the simultaneous mutation of BRAF V600E and TERT promoter. Detection of simultaneous mutations in BRAF V600E and TERT promoters could indirectly predict the iodine uptake ability of DTC tissues. 5. TSHR expression in DTC patients was affected by both BRAF V600E and TERT promoter mutations.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R736.1

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