Clinical and Pathological Characteristics of Extrapleural Solitary Fibrous Tumor
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摘要:
目的 探讨胸膜外孤立性纤维性肿瘤的临床病理特征及鉴别诊断。 方法 对26例胸膜外孤立性纤维性肿瘤样本进行HE及免疫组织化学染色,并结合临床及影像学资料进行综合分析。 结果 孤立性纤维性肿瘤的组织学形态多样,典型者可见卵圆形或梭形细胞不规则排列,间质中可见粗大的胶原带、鹿角状血管及管周透明变性;免疫组化染色STAT-6、CD34、Bcl-2、CD99、SMA阳性率分别为96.2%、92.3%、92.3%、76.9%、26.9%。 结论 SFT常见发生于胸膜,亦可发生于其它部位,其组织学特征和免疫组化表型有助于明确诊断,但当其发生于不常见的解剖部位时,应当对重点需要鉴别或排除的疾病加以认真细致的鉴别并综合判断其临床预后。 -
关键词:
- 胸膜外孤立性纤维性肿瘤 /
- 组织学形态 /
- 生物学行为 /
- 临床预后
Abstract:Objective To investigate the clinical and pathological characteristics of the extrapleural solitary fibrous tumor and the relevant differential diagnosis. Methods HE and immunohistochemical staining were performed on 26 samples of isolated fibrous tumors outside the pleura, and a comprehensive analysis was conducted based on the clinical and imaging data. Results The histological morphology of isolated fibrous tumors was diverse, with typical cases showing the irregular arrangement of oval or spindle shaped cells. In the stroma, thick collagen bands, antler like blood vessels, and peritubular transparent degeneration could be seen. The immunohistochemical staining demonstrated that the positive rates of STAT-6、CD34、Bcl-2、CD99、SMA were 96.2%、92.3%、92.3%、76.9% and 26.9%, respectively. Conclusion SFT commonly occurs in the pleura and can also occur in other areas. Its histological characteristics and immunohistochemical phenotype help to clarify the diagnosis. However, when it occurs in uncommon anatomical sites, it is necessary to carefully and meticulously distinguish and comprehensively judge the clinical prognosis of key diseases that need to be distinguished or excluded.. -
痛风是常见的代谢类疾病,一般由慢性高尿酸血症后尿酸钠晶体在关节内沉积引起[1-3]。痛风和高尿酸血症与肥胖、高血压、高脂血症、冠心病、胰岛素抵抗等代谢综合征密切相关,已成为识别早期代谢综合征的标志[4]。以往的研究表明,高尿酸血症是痛风发生和痛风形成的基础。因此,通过观察高尿酸血症的相关影响因素,可能会预测到痛风发生。越来越多的证据表明,痛风和高尿酸血症在治疗上表现出明显的个体差异性,这主要归因于基因家族某些成员的遗传变异[5]。全基因组关联分析结果显示,ATP转运蛋白G超家族成员2基因(ATP-binding cassette,subfamily Gmember 2; ABCG2)中rs2231142位点、葡萄糖转运基因9(Solute carrier family 2 member 9;SLC2A9)中rs6855911和rs1014290位点、溶质载体家族17成员3(Solute Carrier Family 17 Member 3,SLC17A3)基因中rs1165205位点和cGMP依赖性蛋白激酶2(Protein Kinase CGMP-Dependent 2,PRKG2)基因中rs7688672位点的多态性在高尿酸血症的发生中起着重要作用[6]。基于上述研究背景,本研究通过对云南红河州地区哈尼族和汉族痛风病和高血尿症患者ABCG2、SLC2A9、SLC17A3和PRKG2基因中5个位点的单核苷酸多态性,分析其基因型和等位基因频率是否与该地区痛风和高尿酸血症的发病存在关联。
1. 资料与方法
1.1 研究对象
选择2017年1月至2019年12月至宁洱县中医院确诊的哈尼族痛风患者100例为哈尼族患病组; 100例汉族痛风患者为汉族痛风患病组。选择同期健康体检者哈尼族126例为哈尼族对照组,108例汉族对照组。痛风的纳入标准符合1977年美国风湿病学会制定的痛风标准[7],且排除因恶性肿瘤、血液系统疾病和服用嘌呤酶代谢药物导致尿酸水平变化的患者。该实验得到宁洱县中医院伦理委员会支持,且所有参与者均知情。
1.2 临床指标检测
在空腹状态下,采集所有参与者静脉血3 mL,离心后得到血清,检测血清中可以反映肝功能、肾功能和脂质代谢相关的血液生化指标水平:谷丙转氨酶(alanine aminotransferase,ALT)、门冬氨酸氨基转移酶(aspartate aminotransferase,AST)、尿素氮(blood urea nitrogen,BUN)、血清肌酐(creatinine,CREA)、尿酸(uric acid,UA)、血清总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、高密度脂蛋白(high density lipoprotein cholesterol,HDL-C)和低密度脂蛋白(low density lipoprotein cholesterol,LDL-C)。
1.3 SNaPshot SNP分型
从外周血样本中提取总DNA,SNaPshot多重分析技术对ABCG2(rs2231142)、SLC2A9(rs6855911和rs1014290)、SLC17A3(rs1165205)和PRKG2(rs7688672)这4个基因的5个SNP进行基因分型和筛选,再用PCR技术对ABCG2、SLC2A9、SLC17A3和PRKG2的基因分型进测序和确定。本研究所使用的PCR引物,见表1。
表 1 引物序列Table 1. Primer sequences位点 引物序列 rs2231142-F CTGCAGGTTCATCATTAGCTAGAAC rs2231142-R TTCATGATTCGTCATAGTTGTTGC rs6855911-F TCCCACACTAGCATATAAGAGGAAC rs6855911-R GTGGTCAAGAAAGTGTCCAGCTAC rs1014290-F AATGATACAATGCATTATCAGCTCC rs1014290-R GATGATGAAGTTTCCAGACCTCAG rs1165205-F TGCTAATGGAAACATCATATTCTTG rs1165205-R CATGTGCTTTCATTTCTCTCAGATA rs7688672-F ACTATGACTTTTCTGACAATGGCC rs7688672-R TCCAGAGTGGCTTTAATTGAACAT 1.4 统计学处理
用SPSS 20.0软件对数据进行分析和处理。基因型和等位基因频率用构成比(%)表示。组间用χ2检验进行分析,如果结果不符合χ2检验,则用Fisher确切概率法进行分析。组间比较采用Tukey post-hoc pairwise分析,数据以均值±标准差(Mean±SD)表示。P < 0.05为差异具有统计学意义。
2. 结果
2.1 临床血液生化指标测定比较
本研究共包括四组人群,分别为哈尼族痛风组(100人)、哈尼族对照组(126人)、汉族痛风组(100人)、汉族对照组(108人),共计434人,检测各组肝功能、肾功能和脂质代谢相关的血液生化指标,见表2。结果表明,在哈尼族痛风组和哈尼族对照组的血液生化指标中的ALT、AST、BUM、CREA、UA、 TC、TG、HDL-C、LDL-C的差异有统计学意义(P < 0.05),其他血液生化指标均无统计学意义(P > 0.05)。在汉族痛风组和汉族对照中的血液生化指标中只有CREA、UA、TC、HDL-C、LDL-C的差异存在统计学意义 (P < 0.05)。在汉族组和哈尼族组中,只有LDL-C的差异在哈尼族对照组和汉族对照组中存在统计学意义(P = 0.001)。
表 2 汉族和哈尼族痛风组和非痛风组的个人基本资料和临床生化指标测定比较(mean±SE)Table 2. The clinical features of subjects with gout and non-gout from 226 Hani populations and 208 Han populations in China (mean ± SE)民族
指标哈尼族 汉族 痛风组 (n = 100) 对照组 (n = 126) 痛风组 (n = 100) 对照组 (n = 108) 性别 男/女(n) 78/22 84/42 78/22 69/39 P 0.035 ALT(U/L) 41.17 ± 7.97 27.83 ± 1.97 37.92 ± 4.02 29.14 ± 1.79 F 16.37 3.931 1.211 5.044 P 0.028* 0.612 0.825 0.162 AST(U/L) 43.82 ± 10.94 27.13 ± 1.06 33.56 ± 5.19 26.42 ± 1.04 F 106.5 4.443 1.039 24.90 P 0.034* 0.215 0.927 0.378 BUM(mmol/L) 7.91 ± 1.03 5.91 ± 0.15 6.45 ± 0.41 5.85 ± 0.51 F 47.15 0.0459 11.56 1.547 P 0.013* 6.311 0.941 0.472 CREA(μmol/L) 109.73 ± 9.75 76.00 ± 5.42 112.64 ± 11.93 82.93 ± 4.36 F 3.236 0.0459 1.545 7.487 P 0.005* 0.818 0.555 0.017* UA,(μmol/L) 489.11 ± 9.53 303.99 ± 5.46 523.60 ± 36.98 304.63 ± 7.02 F 3.046 15.06 1.653 27.75 P 0.0002* 0.199 0.980 0.0002* TC(mmol/L) 5.18 ± 0.12 5.14 ± 0.09 5.00 ± 0.10 5.38 ± 0.13 F 1.778 1.440 2.086 1.690 P 0.783 0.247 0.109 0.015* TG(mmol/L) 2.60 ± 0.25 1.78 ± 0.12 2.49 ± 0.18 2.18 ± 0.20 F 4.340 1.929 2.778 1.235 P 0.001* 0.680 0.099 0.258 HDL-C,(mmol/L) 1.25 ± 0.04 1.54 ± 0.04 1.24 ± 0.03 1.47 ± 0.05 F 1.000 1.778 1.563 2.778 P < 0.0002* 0.896 0.213 < 0.0002* LDL-C,(mmol/L) 2.77 ± 0.09 2.82 ± 0.08 2.79 ± 0.10 3.25 ± 0.10 F 1.266 1.235 1.563 1.000 P 0.699 0.927 0.001* 0.001* 注:*P < 0.05。 2.2 哈尼族和汉族基因频率与高尿酸血症的关系比较
分析哈尼族(n = 226)和汉族(n = 208)中5个位点的基因型和等位基因频率,5个位点的基因型分布符合Hardy-weinberg遗传平衡定律,具有群体代表性(P < 0.05)。哈尼族中rs1014290位点基因型频率(P = 0.008)和等位基因频率(P = 0.023)与汉族相比,差异具有统计学意义,其余4个位点差异无统计学意义(P > 0.05),见表3。
表 3 哈尼族和汉族5个SNPs基因不同位点基因型及等位基因频率比较[n(%)]Table 3. Genotype frequencies of five SNPs in Hani populations and Han population [n(%)]基因型 哈尼族(n = 226) 汉族(n = 208) χ2 P 次要等位基因频率 rs2231142 0.119409 GG 99(48.1) 107(51.9) 3.24 0.198 GT 106(56.7) 81(43.3) TT 18(47.4) 20(52.6) G 205(52.2) 188(47.8) 0.319 0.613 T 18(47.4) 20(52.6) rs6855911 0.302915 A 214(52.8) 191(47.2) 3.25 0.104 G 9(34.6) 17(65.4) rs1014290 0.32528 AA 97(58.8) 68(41.2) 9.586 0.008* AG 89(43.8) 114(56.2) GG 37(58.7) 26(41.3) A 97(58.8) 68(41.2) 5.318 0.023* G 126(47.4) 140(52.6) rs1165205 0.30611 AA 148(53.2) 130(46.8) 0.707 0.702 AT 70(49.0) 73(51.0) TT 5(50.0) 5(50.0) A 218(51.8) 203(48.2) 0.012 0.911 T 5(50.0) 5(50.0) rs7688672 0.490216 AA 76(54.7) 63(45.3) 2.58 0.275 AG 110(52.9) 98(47.1) GG 37(44.0) 47(56.0) A 76(54.7) 63(45.3) 0.708 0.411 G 147(50.3) 145(49.7) 注:*P < 0.05。 2.3 哈尼族和汉族高尿酸血症组与5个SNPs位点的相关性
通过分析哈尼族高尿酸血症组(n = 100)和哈尼族对照组(n = 123)中5个SNPs位点的分布可知,在哈尼族中只有rs2231142位点在基因型频率(P = 0.002)和等位基因频率(P = 0.024)的差异与高尿酸血症显著相关,具有统计学意义。此外SNP rs2231142 的TT基因型和T等位基因会增加患高尿酸血症的风险。
分析汉族高尿酸血症组(n = 100)和汉族对照组(n = 108)中5个SNPs位点的分布。结果显示,在汉族中rs7688672位点在基因型频率(P = 0.018)和等位基因频率(P = 0.034)的差异与高尿酸血症显著相关。此外,汉族高尿酸症和rs1165205位点的基因型频率(P = 0.037)显著相关,但是与rs1165205位点等位基因频率无相关性,见表4。
表 4 高尿酸血症组和对照组中ABCG2 (rs2231142),SLC2A9 (rs6855911 和rs1014290),SLC17A3 (rs1165205) 和 PRKG2(rs7688672)基因型频率比较[n(%)]Table 4. Genotype frequencies and associations tests of ABCG2 (rs2231142),SLC2A9 (rs6855911 and rs1014290),SLC17A3 (rs1165205) and PRKG2(rs7688672) in Hyperuricemia patients and non-Hyperuricemia subjects [n(%)]基因型 哈尼族 汉族 高尿酸血症组
(n = 100)对照组
(n = 126)χ2 P 高尿酸血症组
(n = 100)对照组
(n = 108)χ2 P rs2231142 GG 33(33.3) 66(66.7) 12.352 0.002* 44(41.1) 63(58.9) 5.182 0.075 GT 54(50.9) 52(49.1) 43(53.1) 38(46.9) TT 13(72.2) 5(27.8) 13(65.0) 7(35.0) G 87(42.4) 118(57.6) 5.934 0.024* 87(46.3) 101(53.7) 2.539 0.157 T 13(72.2) 5(27.8) 13(65.0) 7(35.0) rs6855911 A 95(44.4) 119(55.6) 0.435 0.519 94(49.2) 97(50.8) 1.212 0.318 G 5(55.6) 4(44.4) 6(35.3) 11(64.7) rs1014290 AA 50(51.5) 47(48.5) 3.191 0.203 36(52.9) 32(47.1) 1.803 0.406 AG 36(40.4) 53(59.6) 50(43.9) 64(56.1) GG 14(37.8) 23(62.2) 14(53.8) 12(46.2) A 50(51.5) 47(48.5) 3.119 0.08 36(52.9) 32(47.1) 0.958 0.376 G 50(39.7) 76(60.3) 64(45.7) 76(54.3) rs1165205 AA 69(46.6) 79(53.4) 1.317 0.517 71(54.6) 59(45.4) 6.569 0.037* AT 28(40.0) 42(60.0) 28(38.4) 45(61.6) TT 3(60.0) 2(40.0) 1(20.0) 4(80.0) A 97(44.5) 121(55.5) 0.475 0.659 99(48.8) 104(51.2) 1.618 0.371 T 3(60.0) 2(40.0) 1(20.0) 4(80.0) rs7688672 AA 32(42.1) 44(57.9) 0.531 0.767 23(36.5) 40(63.5) 8.051 0.018* AG 52(47.3) 58(52.7) 47(48.0) 51(52.0) GG 16(43.2) 21(56.8) 30(63.8) 17(36.2) A 32(42.1) 44(57.9) 0.349 0.573 23(36.5) 40(63.5) 4.845 0.034* G 68(46.3) 79(53.7) 77(53.1) 68(46.9) 注:*P < 0.05。 3. 讨论
与欧美国家相比,20世纪50年代以前,亚洲地区高尿酸血症和痛风的患病率极低,属于罕见病。但在20世纪90年代中期以后,随着经济的发展和生活方式的改变,我国男性高尿酸血症的发病率为5.8%~33.1%,女性为2.4%~11.9%[1]。据研究报道,嘌呤代谢紊乱或者尿酸排泄减少,导致血清尿酸浓度过高,尿酸钠结晶沉积在关节、软骨、肾脏等组织会引发急性痛风关节炎[8]。痛风和高血尿症的研究在世界上不同地区和不同人群中进行,然而在中国,对痛风的研究仅限于汉族[9]。哈尼族是云南25个世居民族之一,有163万人口[10]。哈尼族大部分居住在云南省红河哈尼族彝族自治州和西双版纳傣族自治州,此外,还广泛分布在东南亚(包括越南、泰国、缅甸、老挝)[11]。本研究分析了哈尼族和汉族5个位点的基因型分布,这些发现为痛风和高尿酸血症的遗传效应提供了进一步的证据。
ABCG2,位于人类染色体4q22-q23,编码尿酸代谢的转运蛋白。全基因组关联分析发现,ABCG2是痛风和高尿酸血症的易感基因[12],ABCG2基因rs2231142位点的Q141K突变,导致ABCG2蛋白的稳定性下降而且表达量降低,影响了尿酸的排泄功能,且显著影响痛风的发病年龄,被认为是高尿酸血症和痛风的重要调节因子[13]。在本次研究中,哈尼族组中rs2231142中T等位基因频率会增加患痛风和高尿酸血症的风险,这和文献报道的结果是一致的。Yamagishi报道[14]日本人群中ABCG2基因rs2231142位点T等位基因与较高血尿酸水平和痛风发病率较高有关。这些研究结果显示,rs2231142位点的多态性产生错意突变导致的Q141K,使ABCG2基因第五外显子发生错配,最终导致ABCG2蛋白的失调,从而造成机体高尿酸血症,从而引发痛风[15]。
SLC2A9位于人类染色体4p16.1[16],编码GLUT9蛋白,GLUT9在肾小管上皮细胞和关节软骨细胞中高表达,肾小管上皮细胞参与尿素的重吸收和运输,关节软骨是痛风中尿酸沉积的位置[17]。SLC2A9与尿酸水平和痛风的关系,国内外研究的较多的为rs6855911和rs1014290两个位点[18]。本研究发现,rs6855911和rs1014290位点基因的多态性和哈尼族与汉族的高尿酸血症没有关系。此外,SLC2A9基因突变,使其编码的GLUT9功能缺失,导致2型低尿酸血症[19]。然而全基因组关联分析发现的SLC2A9多态性和与尿酸转运的功能和机制尚不清楚,因此,需要更大的样本量和多民族人群来验证。
SLC17A3基因,编码磷酸钠载体蛋白(Sodium-dependent phosphate transport protein 4,NPT4),NPT4集中分布在肾近端小管细胞膜上端。SLC17A3基因中rs1165205位点与尿酸浓度有关联[20],在笔者的研究中SLC17A3基因中rs1165205位点的基因频率和高尿酸血症相关,而且基因型AA增加了汉族人患高尿酸血症的风险。cGMP依赖性蛋白激酶2(Protein Kinase CGMP-Dependent 2,PRKG2),位于人类染色体4q13.1-21.1区。PRKG2基因编码的蛋白激酶属于丝/苏氨酸蛋白激酶家族,软骨是其主分布区之一,可以促进软骨细胞的增生和肥大,对成骨过程有促进作用。此外,有研究证实,PRKG2基因变异导致蛋白功能异常,且PRKG2基因中rs7688672位点基因的多态性与痛风有关[21]。然而,在以往的研究中,rs7688672位点基因的多态性变异与痛风的关系互相矛盾[22]。本研究发现rs7688672位点中等位基因A能够降低汉族人群患高尿酸血症的风险。
本研究以434例红河州人群样本为研究对象进行分析,发现在哈尼族和汉族痛风和非痛风人群中血液生化指标具有很大的不同。根据本研究的分析结果,LDL-C的表达水平在哈尼族对照组汉族对照组中具有显著差异,其他血液生化指标均没有显著差异。因此,笔者进一步比较了哈尼族组和汉族组中5个SNPs位点,发现SLC2A9基因中rs1014290位点的基因频率和基因型频率具有显著差异。这进一步的证明,在痛风中汉族和哈尼族人群之间存在个体差异。
总之,笔者的研究结果表明,ABCG2基因中 rs2231142位点多态性与哈尼族高尿酸血症的风险有关,而SLC17A3基因中的 rs1165205位点和PRKG2 基因中的rs7688672位点多态性与哈尼族高尿酸血症的风险有关。这些研究结果表明,rs2231142、rs1165205和rs7688672与痛风明显相关,可作为痛风和高尿酸血症筛查、诊断和治疗的潜在候选生物标志物,有助于提高临床上痛风和高尿酸血症患者的个体化治疗水平。但本研究仍然有不足之处,云南作为多民族地区,本研究只纳入哈尼族与汉族人群,未纳入其他少数民族人群进行分析,笔者将进一步研究本研究关注的五个多核苷酸位点与不同族群的痛风和高尿酸血症患者的关系。
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表 1 病例基本情况
Table 1. Basic information of patients
性别 n 年龄(岁) 肿瘤大小(cm) 男 15 >55(n = 6) >5(n = 14) 女 11 ≤55(n = 20) ≤5(n = 12) 表 2 胸膜外孤立性纤维性肿瘤的病理学特征
Table 2. Pathological characteristics of ESFT
病理特征 n 占比(%) 组织学形态 “鹿角状”薄壁血管 13 50.0 血管周透明变性 6 23.1 细胞丰富、有异型性 1 3.8 小灶坏死 1 3.8 间质黏液变性 6 23.1 微囊性变 3 11.5 小出血灶 3 11.5 成熟脂肪细胞 2 7.7 淋巴细胞浸润 2 7.7 免疫表型 STAT-6 25 96.2 CD34 24 92.3 Bcl-2 24 92.3 Vimentin 26 100.0 CD99 20 76.9 SMA 7 26.9 S-100 6 23.1 EMA 13 50.0 Desmin 1 3.8 CD117 0 0 -
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