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甲状腺乳头状癌(PTC)是最常见的甲状腺癌病理类型,约占所有类型的80%。最新调查结果显示,过去的30年间,全球PTC的发病率增加了约58%,因此对于PTC临床和基础研究的价值逐渐凸显[1]。作为一种内分泌肿瘤,PTC受到多种基因影响,尤其是BRAFV600E基因突变被认为是目前最常见的基因变异,在PTC形成过程中发挥重要作用,影响其发生、侵袭及预后,被公认为是PTC最主要的不良影响因子之一[2]。131I是PTC的主要治疗方法,其发挥杀伤肿瘤细胞的效应依赖细胞基底膜表达的钠碘转运体(NIS),以保证细胞必须具有较强的摄碘能力,然而,NIS在PTC组织内的表达是下降或者缺失的,从而导致131I治疗效果不佳,也有研究[3]表明BRAF突变可能影响NIS蛋白表达,因此本文将继续探讨PTC中常见的遗传学事件BRAFV600E突变与NIS表达的相互关系,为PTC病人放射性131I治疗效果的预测提供参考。
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BRAFV600E基因突变检测标本共104例,结果表明,BRAFV600E基因突变型共70例,突变率为67.3%,野生型34例,突变率为32.7%,同样来源于104例PTC观察对象的癌旁正常组织一起进行BRAFV600E基因突变检测,结果提示均为野生型。组间突变率比较差异明显(χ2=105.51, P < 0.01)。
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所提取的总RNA经微量核算定量分析仪检测浓度,结果提示:PTC组织中NISmRNA的表达浓度低于癌旁正常组织,差异有统计学意义(P < 0.01)。与癌旁正常组织相比,NISmRNA表达下调90例(86.5%),14例表达上调(13.5%)。免疫组化检测结果显示,PTC组织中,NIS蛋白阳性表达率高于癌旁正常组织(P < 0.01)(见表 1)。
分组 n NISmRNA (相对表达量) NIS蛋白[n;百分率(%)] 甲状腺乳头状癌组织 104 -3.30(-5.00, 15.27) 77(74.0) 癌旁正常组织 104 3.38(-5.38, 14.88) 45(43.3) χ2 — 2.62* 20.30 P — < 0.01 < 0.01 *示Zc值 表 1 不同甲状腺组织NISmRNA及NIS蛋白表达
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荧光定量PCR检测及免疫组化分析发现,与癌旁正常组织比较,PTC组织中NISmRNA表达下降的同时,有76例NIS蛋白表达上调,14例NIS蛋白表达下调,NISmRNA表达上调时,14例NIS蛋白表达上调,PTC组织中NISmRNA表达下调同时NIS蛋白表达上调的比例明显高于癌旁正常组织。
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PTC BRAFV600E基因突变病人NISmRNA表达浓度为1.57×106copies/mL,明显低于野生型NISmRNA表达浓度1.74×106copies/mL,差异有统计学意义(P < 0.01);PTC BRAFV600E基因突变病人中NIS蛋白表达阳性者49例,阳性率为70.0%,野生型病人中NIS蛋白表达阳性者28例,阳性率为82.4%,组间比较差异无统计学意义(P>0.05)(见表 2)。
分组 n NISmRNA (相对表达量) NIS蛋白[n; 百分率(%)] BRAFV600E基因 70 -4.22(-5.19, 14.83) 49(70.0) 野生型 34 3.73(-5.11, 13.69) 28(82.4) χ2 — 2.34* 1.82 P — < 0.01 >0.05 *示Zc值 表 2 PTCNISmRNA及NIS蛋白表达与BRAFV600E基因突变的关系
甲状腺乳头状癌钠碘转运体表达与BRAFV600E突变的相关性研究
Correlation between sodium iodine transporter expression and BRAFV600E mutation in thyroid papillary carcinoma
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摘要:
目的 探究甲状腺乳头状癌(PTC)中常见的遗传学事件BRAFV600E突变与钠碘转运体(NIS)表达的关系。 方法 选取104例行手术治疗的PTC病人为观察对象,留取术中新鲜PTC组织和癌旁正常组织标本,通过荧光定量PCR法检测NISmRNA含量,石蜡包块行免疫组织化学检测NIS蛋白表达,同时行BRAFV600E突变基因检测。 结果 104例PTC组织中BRAFV600E基因突变型共70例(67.3%),高于癌旁正常组织0例(P < 0.01);PTC组织中NISmRNA的表达浓度低于癌旁正常组织(P < 0.01),但NIS蛋白表达强度则高于癌旁正常组织(P < 0.01);PTC组织中NISmRNA表达下调同时NIS蛋白表达上调的比例高于癌旁正常组织(P < 0.01);PTC BRAFV600E基因突变病人NISmRNA表达浓度低于野生型病人(P < 0.01),但NIS蛋白比较差异无统计学意义(P>0.05)。 结论 BRAFV600E基因突变仅存在于癌组织中,可能与癌组织中NISmRNA低表达及NIS蛋白高表达具有相关性,且推测PTC中NIS蛋白可能是主要表达于细胞质中,影响其正常运输功能而导致病人无法聚碘。 -
关键词:
- 甲状腺乳头状癌 /
- 钠碘转运体 /
- BRAFV600E基因
Abstract:Objective To explore the relationship between BRAFV600E mutation and sodium iodide transporter(NIS) in common genetic events in thyroid papillary carcinoma(PTC). Methods A total of 104 PTC patients who underwent surgical treatment PTC were selected. Fresh PTC tissue and normal tissue adjacent to carcinoma specimens were collected. The contents of NIS mRNA were detected by fluorescence quantitative PCR. NIS protein expressions in paraffin blocks were detected by immunohistochemistry, at the same time BRAFV600E gene mutation was tested. Results In 104 cases of PTC, there were 70 cases of BRAFV600E gene mutation(67.3%), which was significantly higher than that in the normal tissue adjacent to carcinoma(0 case)(P < 0.01). The expression of NIS mRNA in PTC tissues was significantly lower than that in the normal tissues adjacent to carcimoma(P < 0.01), but the expression of NIS protein was significantly higher than that in the normal tissues adjacent to carcimoma(P < 0.01). The proportion of NIS mRNA expression in the PTC tissues was lower and the increase of NIS protein expression was significantly higher than that in the normal tissues adjacent to carcimoma(P < 0.01). The expression of NIS mRNA in patients with gene mutation of PTC BRAFV600E was significantly lower than that in the wild type(P < 0.01), but there was no significant difference in NIS protein(P>0.05). Conclusions BRAFV600E mutation only exists in the cancer tissue, which may be associated with the low expression of NIS mRNA in cancer tissue and high expression of the NIS protein. NIS protein in PTC may be mainly expressed in the cytoplasm, which affects the normal function of transportation and causes patients to be unable to gather iodine. -
Key words:
- thyroid papillary carcinoma /
- sodium iodine transporter /
- BRAFV600E gene
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表 1 不同甲状腺组织NISmRNA及NIS蛋白表达
分组 n NISmRNA (相对表达量) NIS蛋白[n;百分率(%)] 甲状腺乳头状癌组织 104 -3.30(-5.00, 15.27) 77(74.0) 癌旁正常组织 104 3.38(-5.38, 14.88) 45(43.3) χ2 — 2.62* 20.30 P — < 0.01 < 0.01 *示Zc值 表 2 PTCNISmRNA及NIS蛋白表达与BRAFV600E基因突变的关系
分组 n NISmRNA (相对表达量) NIS蛋白[n; 百分率(%)] BRAFV600E基因 70 -4.22(-5.19, 14.83) 49(70.0) 野生型 34 3.73(-5.11, 13.69) 28(82.4) χ2 — 2.34* 1.82 P — < 0.01 >0.05 *示Zc值 -
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