喉鳞状细胞癌中微卫星的异常改变

目的：从分子生物学水平探究微卫星的不稳定性（MSI）与杂合性缺失（LOH）在喉鳞状细胞癌发病机制中的意义。方法：选择3号,5号及11号染色体的3个微卫星位点采用PCR和聚丙烯酰胺凝胶电泳-硝酸银染色方法对40例喉鳞状细胞癌患者手术切除的癌组织及癌旁正常组织进行微卫星分析。结果：40例喉鳞状细胞癌中,35例（87．5％）分别有1～3个微卫星位点发生MSI或LOH。微卫星异常改变发生率最高的位点为D5s592,占70％（28／40）;其次是D3s1228位点,占52．5％（21／40）。结论：在3p14区域及5q23区域附近的抑癌基因参与致癌机制,D3s1228和D5s592的微卫星改变与喉鳞状细胞癌的临床分期相关。     

喉鳞癌组织中RB1.20座位杂合性丢失和p110^RB1蛋白表达 …

目的 探讨喉鳞状细胞癌组中ＲＢ１基因在喉癌发生发展中的意义，为发现和定位新的抑癌基因提供线索和依据。方法 应用聚合酶链反应，选择１３号染色体上ＲＢ１．２０座位的微卫星多态标记，对５８例喉鳞状细胞癌组织进行杂合性丢失分析和ｐ１１０＾ＲＢ１蛋白表达的检测。结果 ３例原位癌中均无ＲＢ１．２０座位杂合性丢失，５５例浸润癌中４５例中提供信息，杂合性丢失的频率为４０％，４１例中仅有８例ＲＢ蛋白表达缺失，其中６     

喉癌三号染色体短臂(3p)抑癌基因微卫星多态位点的杂合性缺失检测

目的 :检测喉鳞癌组织在 3号染色体短臂上 (3p)三个区段 3p14 - cen、3p2 1- 2 2、3p2 5 - 2 6内相应抑癌基因FHIT,h ML H1及 VHL微卫星 (microsatellite,MS)的杂合性缺失 (loss of heterozygosity,L OH)。方法 :采用显微切割法从病理切片中挑取肿瘤组织 ,选取上述基因内含子或与其连锁的 7个 MS多态位点对 4 1例喉癌组织进行聚合酶链式反应和变性凝胶电泳。结果 :FHIT基因内含子三个 MS位点 D3S12 34、D3S130 0及 D3S4 10 3的 L OH发生率分别为77.4 2 %、6 8.97%和 6 9.70 %。与 h ML H1基因连锁的两个 MS位点 D3S15 6 1和 D3S16 12的 L OH发生率分别为 5 8.33%和 4 5 .16 %。与 VHL基因连锁的两个 MS位点 D3S10 38及 D3S12 83的 L OH发生率分别为 4 3.33%和 4 6 .88%。 FHIT基因内含子的 MS,与 h ML H1基因连锁的 MS,与 VHL基因连锁的 MS的 L OH发生率分别为 82 .93% (34/ 4 1) ,6 7.5 0 % (2 7/ 4 0 ) ,70 .0 % (2 8/ 4 0 )。同时在上述三个基因区域内发生某一或某些 MS多态位点 L OH者有 16例 ,占 39%。D3S12 34L OH在吸烟指数≥ 4 0 0的喉癌患者较吸烟指数 <4 0 0者明显频发 ,统计学有显著性差异 (P <0 .0 5 )。结论 :喉癌组织 3号染色体短臂上三个抑癌基因 FHIT,h ML H1及 VHL存在高频率的 L OH提     

喉鳞癌组织中RB1.20座位杂合性丢失和p110RB1蛋白表达的检测

目的探讨喉鳞状细胞癌组织中ＲＢ１基因在喉癌发生发展中的意义，为发现和定位新的抑癌基因提供线索和依据。方法应用聚合酶链反应（ｐｏｌｙｍｅｒａｓｅｃｈａｉｎｒｅａｃｔｉｏｎ，ＰＣＲ），选择１３号染色体上ＲＢ１．２０座位的微卫星多态标记，对５８例喉鳞状细胞癌组织进行杂合性丢失（ｌｏｓｏｆｈｅｔｅｒｏｚｙｇｏｓｉｔｙ，ＬＯＨ）分析和ｐ１１０ＲＢ１蛋白表达的检测。结果３例原位癌中均无ＲＢ１．２０座位杂合性丢失，５５例浸润癌中４５例可提供信息，杂合性丢失的频率为４０％（１８／４５），４１例中仅有８例ＲＢ蛋白表达缺失，其中６例伴有ＲＢ１．２０座位的杂合性丢失。结论ＲＢ１基因在部分喉癌中发生失活；在ＲＢ１．２０座位附近可能还存在与喉癌发生发展密切相关的抑癌基因。     

内淋巴囊乳头状癌和脉络丛乳头状瘤对TTR的不同表达

发生于颞骨的侵入性乳头状瘤临床较少见，近来已被证实肿瘤起源于内淋巴囊上皮或内淋巴管上皮，但也有人认为部分肿瘤起源于脉络丛或异位的脉络丛上皮，然而这两种肿瘤在光镜下形态学表现几乎完全相同。有关的文献报告，TTR（transthyretin）可以作为脉络丛上皮的标志，但又缺乏内淋巴囊乳头状瘤免疫组化研究的资料。为进一步区别这两种起源的乳头状瘤，对正常脉络丛上皮标本10例、脉络丛乳头状瘤5例和正常内淋巴囊上皮标本4例、内淋巴囊乳头状瘤4例进行免疫组化研究，使用TTR、CTK（cytokeratin）、S－100蛋白EMA（epithelialmembran…     

脆性组氨酸三联体基因与鼻腔及鼻窦癌相关性研究

目的 分析脆性组氨酸三联体(fragile histindine triad,FHIT)基因及其蛋白表达在鼻腔及鼻窦癌中的异常改变,探讨其与鼻腔及鼻窦癌发生的相关性.方法 回顾性分析1991～2000年收治的48例原发性鼻腔及鼻窦癌患者.免疫组织化学SP法检测FHIT基因的表达.显微切割、变性高效液相色谱分析检测FHIT基因第8外显子微卫星多态标记D3S1234杂合性缺失发生情况.结果 48例癌组织中FHIT蛋白表达缺失5例(10.4%,5/48),与癌旁组织比较表达减弱16例(55.17%,16/29).腺样囊性癌FHIT蛋白表达强于鳞状细胞癌(P＜0.05).杂合子7例(41.18%,7/17),其中杂合性缺失3例(42.86%,3/7),FHIT蛋白表达减少或缺失2例(66.67%,2/3).结论 FHIT基因失活与鼻腔及鼻窦癌的发生相关,可能是该肿瘤的抑癌基因,杂合性缺失可能与鼻腔及鼻窦癌中FHIT蛋白表达下调有关.     

鼻咽癌染色体3p14的精细等位基因缺失分析

目的：进一步明确鼻咽癌染色体3p14区区域等位基因杂合性丢失（loss of heterozygosity,LOH)的频率与共同缺失区规范,以便分离该区域内与鼻咽癌相关的候选抑瘤基因。方法：选择位于3p14的6个高密度微卫星多态标记,对32例鼻咽癌组织进行LOH分析。结果：71．88％（23／31）的鼻咽癌在至少1个位点发生LOH,丢失频率较高的3个位点是D3S1313（46．43％）、D3S1300（50．0％）和D3S1312（44．44％）,在存在丢失的23例患者中,8例表现为一个连续的非随机的LOH区域,其最小共同缺失区为D3S1313－D3S1312（约3．4个厘摩）。且该区域的缺失与鼻咽癌临床分期、EB病毒感染有明显关系。结论：鼻咽癌在3p14的最小共同缺失区位于D3S1313－D3S1312之间,该区域可能存在一个尚克隆的与鼻咽癌发生发展密切相关的抑瘤基因。     

鼻咽癌染色体3p14的精细等位基因缺失研究

目的 研究鼻咽癌染色体３ｐ１４区域的精细等位基因杂合性丢失（ｌｏｓｓ ｏｆ ｈｅｔｅｒｏｚｙｇｏｓｉｔｙ,ＬＯＨ）情况,并探讨ＬＯＨ与鼻咽癌临床分期、临床病理和ＥＢ病毒（Ｅｐｓｔｅｉｎ－Ｂａｒｒ ｖｉｒｕｓ,ＥＢＶ）感染的关系。方法 采用３ｐ１４区域６个精确高密度的微卫星多态性位点,对３２例患者鼻咽癌组织进行ＬＯＨ分析。结果 ３２例患者中有２３例（７１．９％）在至少１个位点发生ＬＯＨ,丢失频率较高     

听神经瘤病因的分子基因分析

听神经痛（家族遗传性和散发性病例）在临床的发病年龄，病程长短，肿瘤生长的速度与生物学方面存在很多差异。为探索临床的差异与分子清理学变异的相关性，对43例听神经瘤患者的肿瘤标本和周围血细胞标本作了分子基因研究。43例中2例为家庭遗传性，41例为散发性病例。主要方法是寻找在正常细胞中存在肿瘤细胞中丢失的等位基因。并集中在已公认的和假设的具有肿瘤抑制基因存在的染色体3P，5q，11P，17P，17q和22。发现22染色体余合性丢失为39畅，与以往报告的22杨一59杨相似。还发现听神经瘤与人类大多数肿瘤的多阶段性、多基因（包括抑制…     

内淋巴囊减压与切开术疗效对比观察

目的 :观察比较内淋巴囊减压与切开术的疗效。方法 :在全麻下按常规手术方法分别行内淋巴囊减压术 2 3例 ,内淋巴囊切开术 2 2例。结果 :术后随访 4 1例 ,随访时间均在 2年以上 ,其中内淋巴囊减压术 2 1例 ,内淋巴囊切开术 2 0例。 2 1例内淋巴囊减压术患者中 ,A级 1 3例 ,B级 4例 ,C级 3例 ,D级 1例 ,完全控制率为6 1 .9% ;2 0例内淋巴囊切开术患者中 ,A级 1 2例 ,B级 5例 ,C级 2例 ,D级 1例 ,完全控制率为 6 0 .0 %。结论 :内淋巴囊减压与切开术在疗效上无显著差异 ,但内淋巴囊减压术较切开术具有操作简单、安全可靠、并发症少等优点 ,因此在术式选择上我们更偏向于内淋巴囊减压术     

喉鳞状细胞癌中脆性组氨酸三联体基因微卫星的不稳定性和杂合性丢失

目的　探讨脆性组氨酸三联体 (fragilehistidinetriad ,FHIT)基因微卫星不稳定性(microsatelliteinstability ,MSI)和杂合性丢失 (lossofheterozygosity ,LOH)与喉鳞状细胞癌 (简称鳞癌 )发生、发展的关系。方法　采用聚合酶链式反应 简单序列长度多态性 银染技术 ,分析 4 1例喉鳞癌中FHIT基因D3S12 34和D3S130 0位点的MSI及LOH。结果　D3S12 34位点LOH发生率为 4 4 4 % (16 /36 ) ,MSI发生率为 19 4 % (7/ 36 ) ;D3S130 0位点LOH发生率为 36 4 % (12 / 33) ,MSI发生率为 2 4 2 % (8/33)。两个位点总的LOH发生率为 5 2 6 % (2 0 / 38) ,总的MSI发生率为 2 8 9% (11/ 38)。总的LOH发生率与喉鳞癌患者TNM分期、病理分级、淋巴结转移及复发有关 (P <0 0 5 ) ,总MSI发生率与喉鳞癌患者淋巴结转移有关 (P <0 0 5 )。结论　FHIT基因LOH和MSI与喉鳞癌的发生、发展有关 ,并可能为喉鳞癌的早期诊断提供新的途径和依据     

Endolymphatic sac tumors: experience of three cases

Objectives

To describe the clinical features, radiological findings, treatment and outcomes of three cases of endolymphatic sac tumors (ELST).

Methods

Retrospective analysis of three cases of ELST.

Results

The first patient had a large ELST invading the labyrinth after a long history of vertigo. He was recurrence-free 1 year after retrolabyrinthine surgical removal. In the second case, an acute peripheral facial nerve paralysis associated with ipsilateral sensorineural hearing loss led to the diagnosis. A translabyrinthine approach was used to remove the tumor, which recurred three times over 10 years. The third patient was a young woman suffering from von Hippel–Lindau (VHL) disease and referred for a sudden sensorineural hearing loss due to an intralabyrinthine hemorrhage secondary to a 2 mm-large endolymphatic sac-confined ELST. Her hearing was totally lost after the deafness recurred 1 month after this first episode. MRI demonstrated a small bilateral ELST. The patient refused surgery on the deaf side.

Conclusion

ELST are difficult to diagnose due to the wide variety of their presentations. Patients with ELST should be screened for VHL disease. Dural invasion and tumor hypervascularization increase the risk of local recurrences after surgery. Early surgical resection may lead to complete tumor removal and inner ear preservation.     

Tumour staged surgery of endolymphatic sac tumors (ELST)

BACKGROUND: Endolymphatic sac tumours (ELST) have only been known as own tumour entities since 1984. ELST might occur solitarily and sporadically as well as hereditary connected to von Hippel-Lindau disease (VHL). This connection has been observed in 1992 for the first time and confirmed by molecular genetic analyses of the VHL gen. There is no agreement yet concerning diagnostics and therapy. METHODS: Our attempt of classifying this type of tumour is the first one. According to our own experience and to literature, we suggest the following classification: ELST type A is locally confined without erosions of the temporal bone nor infiltration of the subarachnoidal area; ELST type B with bony infiltration of the labyrinth block and clinical hearing loss, and ELST type C with additional infiltration of the sigmoid sinus and the vein of jugular bulb. Preoperative diagnostics are performed according to defined radiological criteria in CT and MRI scans including MR-angiography. RESULTS: In 6 patients, including two with a VHL syndrome, ELST was completely sanitized by stage-compatible surgery, using translabyrintine to infratemporal approaches, according to the tumour classification that we developed. The VII (th) nerve could be saved in all tumour stages, and in stage ELST type A the VIII (th) nerve as well. All patients remained without local recurrence in MRI check during the observation period of 4 to 38 months. CONCLUSION: Our stage-compatible surgery of ELST allows total tumor removal with minor morbidity. In contrast to the antero-, retrosigmoidal and suboccipital approaches, the tumour matrix can be safely removed via transmastoidal approach to exclude local recurrences.     

Microsatellite analysis of recurrent vestibular schwannoma (acoustic neuroma) following stereotactic radiosurgery.

HYPOTHESIS: Genetic and immunohistochemical studies may provide insight into the mechanisms of vestibular schwannoma (VS) recurrence following radiation therapy. BACKGROUND: Stereotactic radiation therapy is an increasingly common alternative to microsurgical resection for the primary management of sporadic VS. The molecular mechanisms associated with recurrent vestibular schwannoma (VS) following radiation therapy are not known. METHODS: Primary or irradiated VS tumors were fresh-frozen at the time of surgical resection and microdissected to undergo DNA extraction. Lymphocytic control DNA was isolated from blood obtained by venipuncture. Paired normal and tumor DNA specimens were analyzed for allelic loss by PCR amplification of polymorphic dinucleotide repeat sequences. Immunohistochemical studies were performed on paraffin-embedded, irradiated surgical specimens. RESULTS: Using 16 polymorphic microsatellite markers, 20 of 26 non-irradiated VS demonstrated loss of heterozygosity (LOH) in at least one locus of chromosome 22q. In contrast, none of the four irradiated recurrent VS demonstrated LOH on chromosome 22q (p = 0.008). No allelic loss was seen in either the primary or irradiated VS utilizing markers mapping to chromosome 10. Deletions on chromosome 10 are seen in both benign and higher-grade meningiomas and intracranial malignancies associated with radiotherapy. Immunohistochemical studies were performed to detect the protein product of the NF2 gene, merlin, in the four irradiated VS. NF2 staining was not observed. CONCLUSION: This study represents the first microsatellite and immunohistochemical analysis of recurrent VS following radiation therapy. Our preliminary observations suggest an alternative mechanism of NF2 inactivation that may correlate with radioresistance in VS.     

微切割喉鳞状细胞癌9p13-23区域微卫星杂合性缺失的研究

目的探讨喉鳞状细胞癌(简称鳞癌)在9p13-23区域微卫星(microsatellite)发生杂合性缺失(1ossofheterozygosity,LOH)的热点.方法采用显微切割法从病理切片中挑取肿瘤组织,选取位于9p13-23区域的13个高多态性微卫星引物对42例喉鳞癌组织进行聚合酶链反应和变性凝胶电泳.结果①42例喉鳞癌在9p13-23区域等位基因LOH的总发生率是97.6%(41/42).在13个微卫星引物中,LOH发生率最高者是位于9p22-23的D9S162(89.5%),其次是位于9p21的D9S171(80.0%).与p16基因紧密连锁的D9S1748的LOH发生率仅50.0%.②等位基因缺失作图分析发现42例喉鳞癌组织在9p13-23上存在2个明显的LOH较小区域,分别位于99211的D9S161～D9S171之间和9p22-23的IFNA和D9S162之间.结论喉鳞癌在9p13-23区域除抑癌基因p16以外可能还存在2个或2个以上候选抑癌基因,这些候选抑癌基因也许和p16一样与喉鳞癌的发生、发展密切相关.     

Endolymphatic sac tumours: surgical management

BACKGROUND: Endolymphatic sac tumours (ELSTs) have been known as an individual tumour entity only since 1984. ELSTs may occur either solitarily and sporadically or as a hereditary manifestation associated with von Hippel-Lindau (VHL) disease. The latter association was first observed in 1992 and confirmed by molecular genetic analysis of the VHL gene. No consensual diagnostic and treatment strategy of ELST exists at present. METHODS: Based on imaging criteria in computed tomography, magnetic resonance imaging (MRI), and magnetic resonance angiography, we developed a staging system to classify ELST in a series of seven consecutive patients in an attempt to custom-tailor the surgical approach. Type A referred to tumours that were locally confined without temporal bone erosion or infiltration of the dura (n = 2); type B tumours showed evidence of bone infiltration of the osseous labyrinth and sensorineural hearing loss (n = 2); and in type C, the tumour further invaded the sigmoid sinus and jugular bulb (n = 3). Two patients suffered from VHL disease. RESULTS: In all patients, the tumour was completely removed. Stage-adapted surgical approaches included various transpetrosal procedures, from the translabyrinthine to the infratemporal approaches. The functional integrity of the facial nerve was maintained in all tumour stages, whereas the vestibulocochlear nerve could be preserved only in patients with type A tumours. Follow-up MRI demonstrated no local tumour recurrence during a postoperative observation period ranging from 4 to 38 months. CONCLUSION: Stage-based surgical strategy enables the complete removal of ELST with minor morbidity. Transmastoid approaches are most efficient for resection of the tumour matrix to prevent local recurrence.     

鼻咽癌染色体3p14的精细等位基因缺失研究

目的　研究鼻咽癌染色体 3p14区域的精细等位基因杂合性丢失 (lossofheterozygosity,LOH)情况 ,并探讨LOH与鼻咽癌临床分期、临床病理和EB病毒 (Epstein Barrvirus,EBV)感染的关系。方法　采用 3p14区域 6个精确高密度的微卫星多态性位点 ,对 32例患者鼻咽癌组织进行LOH分析。结果　 32例患者中有 2 3例 ( 71 9% )在至少 1个位点发生LOH ,丢失频率较高的 3个位点是D3S1313( 46 4% )、D3S130 0 ( 5 0 0 % )和D3S1312 ( 44 4% )。在具有丢失的 2 3例患者中 ,12例表现为一个连续的非随机的LOH区域 ,其最小共同缺失区为D3S1313～D3S1312。该区域的LOH与临床分期、EBV感染有明显关系。 30例低分化鳞癌的LOH频率为 70 0 % ,2例泡状核细胞癌均存在 2个位点的LOH。结论　鼻咽癌染色体 3p14区存在较高的LOH率 ,提示在D3S1313和D3S1312之间可能存在尚未克隆的与鼻咽癌发生发展相关的抑癌基因。