骨肉瘤成骨细胞表型与临床因素的相关性研究

目的采集手术切除未经培养的骨肉瘤标本,研究肿瘤中成骨细胞的分化及其与临床因素的关系。方法观察骨肉瘤中成骨细胞相关基因表达的频率、特异性,以及骨肉瘤成骨细胞表型与生物学行为的关系。应用RT-PCR方法研究48例骨肉瘤标本的碱性磷酸酶(ALP)、骨钙素(OC)和骨桥蛋白(ON)的mRNA转录。磷酸甘油酸激酶(PGK)被用于内参照。结果在大多数骨肉瘤标本中可以发现ALP或OC的mRNA转录(分别为93.75%,79.17%)。在所有的肉瘤检测中均可发现ON的mRNA转录。在骨肉瘤的成骨细胞亚型中可见到ALP和OC的高水平表达。肺转移瘤OC的mRNA表达弱于原发灶。肿瘤细胞有高水平OC表达的病例组有较高的生存率(P=0.045)。结论OC是目前最好的成骨细胞特异性标记物,将来可能用于检测骨肉瘤患者外周血中循环的肿瘤细胞。     

siRNA联合顺铂抑制骨肉瘤细胞增殖的研究

目的构建人Survivin基因的siRNA真核表达载体,探讨其对骨肉瘤细胞MG63凋亡及顺铂化疗的增敏作用。方法应用pSilencer 3.0-H1 neo,构建Survivin特异性的RNA干涉载体,转染MG63细胞,G418筛选稳定转染的细胞系。半定量PCR和免疫荧光染色分别检测Survivin mRNA与蛋白的水平变化。Annexin V法检测细胞凋亡。MTT法检测顺铂对细胞的杀伤作用。结果构建Survivin基因siRNA真核表达载体PsiS。获得了稳定转染的细胞系。与MG63、阴性对照细胞比较,MG63/PsiS生长曲线则十分平缓。MG63/PsiS细胞中Survivin的mRNA及蛋白表达明显减少。MG63/PsiS凋亡率增加为17.8%(P<0.01)。1 mg/L顺铂作用下,MG63/PsiS死亡率是其他各组的2.3倍(P<0.01)。结论特异性siRNA能够明显阻断Survivin基因的表达促进MG63细胞凋亡,提高细胞对顺铂的敏感性。     

Dedifferentiated chondrosarcoma

We reviewed 74 cases of dedifferentiated central and peripheral chondrosarcoma. Histologically these tumours consist of an underlying cartilaginous component (either benign or malignant) juxtaposed to a highgrade non-cartilaginous component, with a typically abrupt transition between the two tissue types. The noncartilaginous component may constitute a very small or a very large proportion of the tumour, so diagnosis often requires histological evaluation of the entire tumour. The diagnosis is often suspected on the basis of the clinical course and careful evaluation of the radiographie characteristics. Central dedifferentiated chondrosarcoma can be classified radiographically into three types. In type 1 (36 cases in our review) the radiographie features are the same as those of a central chondrosarcoma, with the addition of a region with very aggressive radiographie features. Type 2 lesions (20 cases) resemble the underlying benign enchondroma but also have destructive changes and/or a large soft tissue mass. Type 3 lesions (8 cases) are not distinctive radiographically and present as a very high grade destructive lesion of bone. These cases are diagnosed following biopsy or tumour resection. The prognosis of these tumours is extremely poor, with 13% overall 5-year survival in this series. Improved survival was found in those cases where diagnosis was prompt and surgical treatment with a wide or radical margin was attained. No benefit was found from the use of adjuvant chemotherapy or radiotherapy. Thus, early recognition of the characteristic radiographie features, adequate histological sampling, and wide or radical surgical margins are necessary for satisfactory management of this highly malignant variant of chondrosarcoma.     

三氧化二砷抑制骨肉瘤细胞及二倍体细胞增殖的研究

目的　研究三氧化二砷 (As2 O3 )对骨肉瘤细胞及二倍体细胞增殖的影响。方法　应用噻唑蓝 (MTT)法和集落形成能力测定、形态学观察、流式细胞术和电镜观察等方法检测和观察As2 O3 对骨肉瘤细胞株MG 63及二倍体细胞株MRC 5增殖的影响。结果　As2 O3 对骨肉瘤细胞及二倍体细胞株的增殖有抑制作用 ,并具有时间依赖性和一定范围内的剂量依赖性。≥ 1μmol/L浓度的As2 O3 对MG 63骨肉瘤细胞的抑制率达 70 % ,而≥ 8μmol/L浓度的As2 O3 对MRC 5细胞的增殖才达到相近的抑制率 ;0 .5～ 2 .0 μmol/L浓度的As2 O3 对MG 63细胞的生长抑制率高 ,而对MRC 5细胞的生长抑制率低 ,表现为明显的选择性抑制 ( P <0 .0 1)。 1μmol/LAs2 O3 处理的MG 63细胞第 3天呈典型的凋亡形态学改变 ;流式细胞仪分析显示 ,在G1期细胞前出现明显亚二倍体峰 ,凋亡率与As2 O3 处理时间呈正相关 ;MRC 5细胞则未见明显凋亡峰。结论　As2 O3 通过诱导细胞凋亡抑制骨肉瘤细胞和二倍体细胞增殖 ,在一定浓度内As2 O3 可选择性抑制骨肉瘤细胞的生长增殖。     

原发性骨小细胞肿瘤的病理形态和免疫组化分析

目的：探讨免疫组化染色及病理形态在原发性骨小细胞肿瘤鉴别诊断中的意义。方法：对39例骨的小细胞肿瘤[其中尤因肉瘤/外周性原始神经外胚层肿瘤（EW/PNET瘤）22例、小细胞骨肉瘤6例、间叶性软骨肉瘤3例和恶性淋巴瘤8例]，观察其病理形态并作免疫组化分析，所用抗体O13、NSE、S-100、Actin和LCA。结果：EW/PNET瘤20/22例为O13阳性。NSE:EW/PNET瘤16/22例和1例骨肉瘤阳性。7例EW/PNET瘤、1例骨肉瘤及3例间叶性软骨肉瘤均有S-100阳性。所有恶性淋巴瘤的LCA均为阳性。骨肉瘤均见骨样基质，间叶性软骨肉瘤可见分化较好的软骨岛。结论：O13的染色对于EW/PNET瘤有诊断意义。小细胞骨肉瘤的成骨和间叶性软骨肉瘤的软骨分化为形态上的诊断要点。     

SIX2在骨肉瘤组织中表达及其对细胞血管生成与阿霉素耐药逆转的影响

探究SIX2在骨肉瘤组织中表达及其对细胞血管生成与阿霉素耐药逆转的影响。方法 收集2019年3月—2021年12月我院行骨肉瘤手术切除的105例患者的组织标本,免疫组化检测骨肉瘤组织中SIX2表达和微血管密度（MVD）计数;检测骨肉瘤组织中SIX2 mRNA表达水平。将人骨肉瘤细胞系MG-63分为MG-63组、si-NC A组、si-SIX2 A组,将通过阿霉素耐药得到的骨肉瘤阿霉素耐药细胞株MG-63/R分为MG-63/R组、si-NC B组和si-SIX2 B组。分别用MTT法、克隆实验、流式细胞术及Transwell法检测细胞耐药性、细胞增殖、细胞凋亡和细胞侵袭。结果 在骨肉瘤组织中,SIX2阳性表达会随着MVD值升高而增加（P＜0.05）。骨肉瘤组织中SIX2阳性表达与TNM分期、软组织浸润和淋巴结远处转移相关（P＞0.05）。与MG-63组相比,si-SIX A组细胞的血管形成能力明显降低（P＜0.05）;与MG-63/R组相比,si-SIX2 B组细胞IC50、细胞克隆数和细胞侵袭能力均显著降低,细胞凋亡能力明显增加,细胞耐药指数逆转倍数为2.51倍（P＜0.05）。结论 SIX2在骨肉瘤组织中呈高表达,且随着SIX2表达的升高肿瘤血管生成能力也增加;抑制SIX2表达可有效抑制骨肉瘤组织的生成,逆转骨肉瘤细胞阿霉素耐药     

Intracisternal crystalline arrays of coated parallel tubules in cells of a human osteosarcoma

Summary Crystalline arrays of coated parallel tubules (CPT) were observed by electron microscopy within dilated cisternae of the rough endoplasmic reticulum of pleomorphic mononuclear cells in a human osteosarcoma. The wall of the peculiar intracisternal tubules consisted of an electron-dense thin membrane-like envelope which appeared to be composed of granular subunits. The electron-lucent tubular core together with the limiting envelope was approximately 15 nm in diameter. A coat of fuzzy material, approximately 10 nm thick, tightly surrounded the membrane-like wall of the tubules. Cross sections of accumulations of CPT showed the tubules to be arranged in a hexagonal crystalline array. The nature and significance of the intracisternal CPT are unknown.     

HIGH-GRADE SURFACE OSTEOSARCOMA OF THE LEFT ILIUM A Case Report and Review of the Literature

A rare case of high-grade surface osteosarcoma of the left ilum is reported. Trephine biopsy performed on a 31-year-old woman suffering from a huge tumor of the left buttock revealed high-grade osteosarcoma, and hemipel-vectomy was undertaken. The surgical specimen showed that the tumor was present on the surface of the left ilium. Ten months after the operation, the patient died of brain metastasis. From these results, we considered this case to be one of high-grade surface osteosarcoma. ACTA PATHOL JPN 38: 235 -240, 1988.     

长链非编码RNA Linc00472靶向微小RNA-381对骨肉瘤细胞增殖、凋亡、迁移、侵袭的影响

目的观察长链非编码RNA Linc00472(LncRNA Linc00472)靶向调控微小RNA-381(miR-381)对骨肉瘤细胞增殖、凋亡、迁移、侵袭的影响。方法收集2017年3月至2018年5月郑州大学第一附属医院收治的骨肉瘤患者29例为研究对象,实时荧光定量聚合酶链反应(RT-qPCR)检测骨肉瘤组织和瘤旁组织中Linc00472的表达。将骨肉瘤U2OS细胞分为pcDNA3.1组、pcDNA3.1-Linc00472组、pcDNA3.1-Linc00472+miR-NC组、pcDNA3.1-Linc00472+miR-381组。甲基噻唑基四唑(MTT)检测细胞增殖;流式细胞术检测细胞凋亡;Transwell实验检测细胞迁移及侵袭。双荧光素酶报告实验鉴定Linc00472与miR-381的靶向关系。两组间比较采用独立样本t检验,多组间比较采用单因素方差分析。结果Linc00472在骨肉瘤组织中的表达水平低于瘤旁组织(0.31±0.03比1.03±0.10,t=37.138,P<0.05),差异有统计学意义;与pcDNA3.1组比较,pcDNA3.1-Linc00472组细胞存活率[(100.29±10.12)%比(53.29±5.44)%]低于pcDNA3.1组(t=12.272,P<0.05),差异有统计学意义,迁移细胞数[(266.00±23.61)个比(124.00±12.01)个]与侵袭细胞数[(131.00±13.06)个比(62.00±6.24)个]少于pcDNA3.1组(t=16.082,P<0.05;t=14.301,P<0.05),差异有统计学意义,而细胞凋亡率[(8.58±0.91)%比(26.61±2.64)%]高于pcDNA3.1组(t=19.370,P<0.05),差异有统计学意义;miR-381过表达可抑制野生型载体WT-Linc00472细胞的荧光素酶活性(t=19.827,P<0.05),差异有统计学意义;与pcDNA3.1-Linc00472+miR-NC组比较,pcDNA3.1-Linc00472+miR-381组可增强细胞增殖[(53.17±5.33)%比(85.26±8.62)%]、迁移[(122.00±12.31)个比(223.00±22.18)个]及侵袭[(64.00±6.36)个比(104.00±10.20)个]能力高于pcDNA3.1-Linc00472+miR-NC组(t=9.499,P<0.05;t=11.945,P<0.05;t=9.983,P<0.05),细胞凋亡率[(26.11±2.62)%比(13.11±1.34)%]低于pcDNA3.1-Linc00472+miR-NC组(t=13.253,P<0.05),差异有统计学意义。结论LncRNA Linc00472通过靶向调控miR-381可抑制骨肉瘤细胞增殖、迁移及侵袭并诱导细胞凋亡。