MicroRNA-26a inhibits osteosarcoma cell proliferation by targeting IGF-1

There are still controversies about the roles of microRNA-26a（miR-26a）in human malignancies,as it is a tumor suppressor in breast cancer,gastric cancer,and hepatocellular carcinoma,but is an oncogene in glioma and cholangiocarcinoma.Until now,the function of miR-26a in osteosarcoma remains largely elusive.Here,we found that miR-26a was downregualted in osteosarcoma tissues.Using in vitro and in vivo assays,we confirmed that miR-26a could inhibit the abilities of in vitro proliferation and suppress in vivo tumor growth in mouse model.Furthermore,we identified insulin-like growth factor 1（IGF-1）as a novel and direct target of miR-26a and revealed that miR-26a exerted its tumor-suppressor function,at least in part,by inhibiting IGF-1expression.These findings contribute to our understanding of the functions of miR-26a in osteosarcoma.

EFFECT OF SURVIVIN-siRNA-MEDIATED GENE SILENCING ON SURVIVIN EXPRESSION IN OSTEOSARCOMA CELL LINE MG-63

Objective： To study the inhibition of expression of Survivin gene by synthesized short Survivin-siRNA in osteosarcoma cell line MG-63. Methods： Chemically synthesized short Surviving-siRNA was transfected into osteosarcoma cell line MG-63. The Survivin mRNA and protein level were detected by reverse transcription-polymerase chain reaction （RT-PCR） and flow cytometry （FCM）. The biological morphology and growth inhibition of MG-63 were observed with light microscopy and MTT assay, respectively. Results： Short siRNA targeting Survivin down-regulated the transcription and the protein level of Survivin oncogene. The proliferation of osteosarcoma cell line MG-63 was inhibited after transfection. Conclusion： Chemically synthesized short Survivin-siRNA can effectively inhibit Survivin expression and cell proliferation inosteosarcoma cell line MG-63 Survivin-siRNA mediated Survivin gene silencing may be a useful therapeutic strategy for osteosarcoma.

EXPRESSION AND SIGNIFICANCE OF ONCOPROTEIN p16 AND FOS IN OSTEOSARCOMA

Objective: To investigate p16, C-fos protein expression and their relationships in osteosarcoma. Methods: Immunohistochemical technique (SABC) was used to detect p16 and C-fos protein expression in 41 cases of osteosarcoma. Results: The positive rates of p16 and C-fos protein expression were 51.2% and 82.9% respectively. Their expression was not correlated to pathological subtype, but correlated to clinic grade, and the latter was associated with tumor metastasis. There was a negative correlation between p16 and C-fos protein expression. Conclusion: The alteration of p16 and C-fos protein expression may be related to the tumorigenesis and development of osteosarcoma, and C-fos proteins may take part in osteosarcoma metastasis. These data will offer useful helpness to determine the prognosis of osteosarcoma.

EXPRESSION OF CYCLIN D1 AND CDK4 IN OSTEOSARCOMA OF THE JAWS

Objective: To analyze cyclin D1 and cyclin-dependent kinase 4 (CDK4) expression and their significance in osteosarcoma of the jaws. Methods: Immunohistochemical ABC method was used to detect the expression of cyclin D1 and CDK4 in 20 cases of osteosarcoma and 8 cases of osteochondroma of the jaws. Results: The positive rates of cyclin D1 and CDK4 were 65% (13/20) and 60% (12/20), respectively. There was significant positive correlation between cyclin D1 and CDK4 expression (γs=0.48, P<0.05). Both cyclin D1 and CDK4 were present in 1/8 (12.5%) osteochondroma. The positive rate was remarkably different between osteosarcoma and osteochondroma (P<0.05). Conclusion: Cyclin D1 and CDK4 are overexpressed in osteosarcoma of the jaws and closely related to its occurrence and development.

Relationship between expression of cathepsin D and proliferating cell nuclear antigen and prognosis in osteosarcoma

Objective: To explore the relationship between expression of cathepsin D (Cath-D) and tumor cell proliferative activity and prognosis in osteosarcoma. Methods: Expression of proliferating cell nuclear antigen (PCNA) and Cath-D in tumor cells were semiquantitatively anajyzed with immunohistochemistry in a consecutiv e prospective series of 85 cases of osteosarcoma. Results: In 85 cases of osteosarcoma, the Cath-D LI was ranged from 6% - 98. 5% and PCNA LI from 9% - 98%. There was no correlation between the Cath-D LI and the patients’ age, sex, tumor size and Dahlin’s classification. However, significant correlation was found between the Cath-D LI and new WHO classification in 1993, Price’s grade and the PCNA LI. Furthermore, Kaplan-Meier analysis showed that the patients with a Cath-D LI less than 66% had a better survival than those with that more than 66%. Conclusion: There is significant relationship between Cath-D expression and survival, suggesting that Cath-D LI might be a useful prognostic indicator in osteosarcoma.

Histopathological and immunohistochemical studies of 120 cases of osteosarcoma

Ｈｉｓｔｏｐａｔｈｏｌｏｇｉｃａｌａｎｄｉｍｍｕｎｏｈｉｓｔｏｃｈｅｍｉｃａｌｓｔｕｄｉｅｓｏｆ１２０ｃａｓｅｓｏｆｏｓｔｅｏｓａｒｃｏｍａＧａｏＦｅｎｇｘｕｎ（高奉浔）；ＬｉＺｅｎｇｐｅｎｇ（李增鹏）；ＤｏｎｇＺｈｉｑｉａｎ（董志谦）；ＳｕＨｏｎｇ...

Expression and distribution of c-fos oncogene within central nervous system of the rat following halothane anesthesia

Objective: To study the distribution of c fos oncogene expression within central nervous system (CNS) of the rat during halothane anesthesia. Methods: c-fos oncogene immunohistochemical technique (ABC method) was employed. Results: When halothane concentration was 0.75%,1.5%or2.0%, most of the Fos-like immunore- active neurons (FLNs) appeared in telencephalon, diencephalon and brain stem, including cerebral cortex, amygaloid nucleus, accumbens nucleus, lateral keptal nucleus. bed nucleus of the stria terminalis, field CA1 of Ammon’s horn, islands of Calleja, paraventricular thalamic nucleus, central medial thalamic nucleus, reuniens thalamic nucleus, rhomboid thalamic nucleus. ventral posterolateral thalamic nucleus, paraventricular hypothalamic nucleus(ventral part). periventricular hypothalamic nucleus, median preoptic nucleus, supraoptic nucleus, suprachiasmatic nucleus. medial and lateral habenular nucleus. midbrain periaqueductal gray and Edinger-Westphal nucleus.In the present stude. we have also found that the number of FLN registered stable increase along with the increaseof the concentration of halothane. Conclusion: It has been indicated that FLNs participate in the process ofhalothane anesthesia. which should necessitate and pave the way for a further study of the patterns of linkage andthe mechanism of interaction between functional nuclei.

Expression and clinical significance of OPN and COX-2 in osteosarcoma

Objective:To detect the expression of osteopontin(OPN)in benign and malignant bone tumors and investigate the prognostic influence of OPN expression on the outcome of osteosarcoma patients.Methods:Fifty-seven osteosarcomas and 11 osteoblastomas as well as 5 bone specimens with remodeling sites were immunohistochemically analyzed for expression of OPN and cyclooxygenase-2(COX-2).Results:OPN was not detected in osteoblasts of remodeling sites.Osteo- blastoma osteoblasts as well as osteoclastlike giant cells and osteosarcoma mononuclear cells showed variable staining.In osteosarcomas OPN and COX-2 expression correlated with each other(r=0.390,P=0.003,Spearman’s test).Although osteosarcoma patients with high COX-2 expression showed a trend towards shorter overall survival(P=0.0841,log-rank test), OPN expression had no influence on patients overall or on disease-free survival.Conclusion:Our data indicated that expres- sion of this protein might be unregulated in bone neoplasia.Although OPN expression correlates with COX-2 expression in osteosarcomas,OPN expression does not provide predictive information about the outcome of osteosarcoma patients.

Tumor blood vessels formation in osteosarcoma:vasculogenesis mimicry

BACKGROUND: Osteosarcoma is characterized by high neovascularization and a high propensity for metastasis through bloodstream. This study was to examine whether there is evidence for vasculogenic mimicry in osteosarcoma and to illustrate mechanism of tumor blood vessels formation in osteosarcoma. METHODS: Osteosarcoma cell lines (U-2OS) were tested for their ability to form tubular networks in three-dimensional culture containing type I collagen. The structures of the tubular networks were observed with phase contrast microscope and transmission electron microscope (TEM). Morphometric studies using hematoxylin and eosin (HE) stain and CD31 immunohistochemical stain to show tumor-lined channels in human osteosarcoma were also performed. RESULTS: Observation with light microscope and TEM showed that highly aggressive osteosarcoma cell lines (U-2OS) formed networks containing channels when grown in three-dimensional culture containing type I collagen, in the absence of endothelial cells or fibroblasts. Morphometric observation using HE stain and CD31 immunohistochemical stain showed that tumor cell-lined channels were also detected in vivo in osteosarcoma; by comparison, all vascular areas in the pedicle of osteochondroma or outside osteochondroma were endothelial-lined. CONCLUSION: These observations strongly suggest that aggressive osteosarcoma cells may generate vascular channels that facilitate tumor perfusion independent of tumor angiogenesis and have the ability of vasculogenic mimicry.

Expression of heat shock protein gp96 in osteosarcoma and its clinical significance

This study aims to detect the expression of heat shock protein gp96(HSPgp96)in osteosarcoma and explore its role in anti-tumor immunity.Expression of HSPgp96 in 45 cases of osteosarcoma,including 24 cases of the osteoblast type,21 cases of chondroblast type and 5 cases of normal bone tissue were detected by immuno-histochemistry(IHC).The expression rate of HSPgp96 was 100%(45/45)in osteosarcoma,compared with 20%(1/5)in normal bone tissue.The average positive cell rates were 97.23%and 0.48%in osteosarcoma and normal bone tissue,respectively,and the difference between them was significant(P<0.01).The positive cell rates in osteoblast type and chondroblast type were 97.78%and 96.56%,respectively,and the difference had no statistical signifi-cance(P>0.05).The expression degree of HSPgp96 in osteosarcoma had a clear correlation with the pathologic degree(P<0.05),while it had no relationship with the clinical stage(P>0.05).The expression of HSPgp96 is high in osteosarcoma and it may be a significant index for judgment of malignancy degree and an important tar-get site of anti-tumor immunity.

Distribution and significance of oncogene C－erbB－1 and suppressor gene р53 in human bladder cancer

To study the relationship between oncogene C-erbB-1 and suppressor gene p53 and bladder cancer,oncogene C-erbB-1 and suppressor gene p53 were determined by immunohistochemistry in 58 cases of bladder cancer. In addition,on 30 of them in situ hybridization

CDC6、DEK、β-catenin蛋白在结直肠癌中的表达及相关性研究

目的分析结直肠癌组织中CDC6、DEK、β-catenin蛋白的表达及临床病理性质,并探讨三者表达的相关性。方法选取结直肠癌(实验组)及对应的癌旁黏膜组织(对照组),每组52例。免疫组化检测2组CDC6、DEK、β-catenin蛋白水平,探讨其与患者临床病理性质的关联,并研究它们之间的相关。结果CDC6、DEK、β-catenin蛋白的表达在实验组阳性率分别为80.77%(42/52)、84.62%(44/52)、65.38%(34/52),在对照组阳性率分别为3.85%(2/52),7.69%(4/52),23.08%(12/52),差异有统计学意义(P<0.01);结肠癌中,CDC6、DEK、β-catenin蛋白高表达均与结肠癌的T分期、临床分期、脉管侵犯呈相关性(P<0.05),且DEK蛋白的表达还与结肠癌分级呈相关性(P<0.05),三种蛋白相互之间具有正相关性。结论CDC6、DEK、β-catenin表达增高与结直肠癌的进展有关,且互相存在正相关,可用作判断结直肠癌预后的潜在参数和药物靶点。

转铁蛋白受体1和血管内皮生长因子在儿童及青少年骨肉瘤患者中的表达及临床意义

目的 探讨转铁蛋白受体1(TFR1)和血管内皮生长因子(VEGF)在儿童及青少年骨肉瘤患者中的表达及临床意义。方法 收集35例儿童及青少年骨肉瘤患者的骨肉瘤组织和癌旁正常组织,采用免疫组化染色法检测TFR1、VEGF表达情况。比较骨肉瘤组织和癌旁正常组织中TFR1和VEGF表达情况,比较不同临床特征儿童及青少年骨肉瘤患者骨肉瘤组织中TFR1和VEGF表达情况,比较不同TFR1、VEGF表达情况儿童及青少年骨肉瘤患者的生存情况,采用Spearman相关分析法分析TFR1与VEGF表达的相关性。结果 儿童及青少年骨肉瘤患者骨肉瘤组织中TFR1和VEGF中高表达率均明显高于癌旁正常组织,差异均有统计学意义(P﹤0.01)。中低分化、有远处转移、Enneking分期为Ⅱ~Ⅲ期儿童及青少年骨肉瘤患者骨肉瘤组织中TFR1、VEGF中高表达率均高于高分化、无远处转移、Enneking分期为Ⅰ期的患者,差异均有统计学意义(P﹤0.05)。TFR1、VEGF低表达儿童及青少年骨肉瘤患者的生存情况均优于TFR1、VEGF中高表达患者,差异均有统计学意义(P﹤0.05)。儿童及青少年骨肉瘤患者中TFR1与VEGF的表达呈正相关(r=0.669,P﹤0.05)。结论 TFR1和VEGF在儿童及青少年骨肉瘤患者中表达上调,可能参与骨肉瘤的发生、发展、侵袭和远处转移,可作为骨肉瘤诊断、恶性程度判断及预后评估指标。

Pim-1在结核性胸膜炎和肺癌胸膜组织中的表达及临床意义

目的探讨Pim-1在结核性胸膜炎及在肺癌患者胸膜组织中的表达及临床意义。方法收集我院2021年1月-2021年11月内科胸腔镜手术切除的结核性胸膜炎及肺癌的胸膜活检标本共41例,其中结核性胸膜炎21例,肺癌20例。利用免疫组织化学法和Western-Blot法分别测定胸膜组织中Pim-1蛋白的表达情况,同时探寻与Pim-1表达有关的临床因素。结果Pim-1在结核性胸膜炎组中的阳性率为95.24%(20/21),在肺癌组中的阳性率为70%(12/20),结核性胸膜炎胸膜组织Pim-1表达水平更高(P<0.001);Western-Blot结果显示Pim-1在结核性胸膜炎中表达水平也高于肺癌组(P<0.01);结核组Pim-1蛋白表达与性别、吸烟史、BMI指数均无明显相关性(P>0.05),肺癌组中Pim-1蛋白表达与性别、吸烟史、BMI指数均无明显相关性(P>0.05),可能与淋巴结转移有关(P<0.01)。结论结核性胸膜炎胸膜组织中的Pim-1表达较高,检测胸膜组织中Pim-1对结核性胸膜炎和肺癌有鉴别诊断价值。

肺癌癌基因蛋白产物同步检测的对比分析

用免疫组织化学方法对７４例肺癌连续切片组织同步进行了ｒａｓ、Ｃ－ｍｙｃ、Ｐ５３三种癌基因蛋白产物的检测。结果显示ｒａｓＰ２１、Ｃ－ｍｙｃＰ６２、和Ｐ５３阳性表达率分别为７１．６％、６４．８％和３３．８％。而癌旁支气管粘膜及腺体分别为１６．１％、２２．６％和０。Ｐ２１和Ｐ６２之间的阳性率无统计学差别（Ｐ＞０．０５），表明这二个癌基因在肺癌发生发展中具有同等的重要性。有两种以上癌基因蛋白同时表达阳性者有４６例（６２．２％），１０例（２１．７％）同时有Ｐ２１、Ｐ６２和Ｐ５３共同表达，结果提示肺癌在发生发展过程中多需要两种或两种以上癌基因作用的结果。

癌基因表达对骨肉瘤患者生存率及生活质量的意义

目的目前有关骨肉瘤中跨膜糖蛋白(ErbB2)表达与患者预后的关系的意见尚不一致,对此做进一步的研究。方法采用免疫组化SABC法检测69例骨肉瘤标本中ErbB2表达,并对有关临床及病理指标综合分析。结果骨肉瘤中ErbB2表达阳性率49.3%。大肿瘤组较小肿瘤组ErbB2表达阳性率高;III期骨肉瘤组ErbB2表达阳性率较II期骨肉瘤组高;ErbB2表达阳性组更容易出现肺转移,并且其生存率降低。结论ErbB2在促进骨肉瘤的侵润性生长和促进转移方面起重要的作用,其阳性表达与恶性程度有关。检测ErbB2的表达情况可以预测骨肉瘤的预后。

ERK5和MMP-9在骨肉瘤中的表达及其临床意义

目的:探讨细胞外信号调节激酶5(extracellular signal-regulated kinase 5,ERK5)蛋白和基质金属蛋白酶-9(matrix metallo proteinase-9,MMP-9)在骨肉瘤组织中的表达及其临床意义。方法:选取71例骨肉瘤组织和40例正常骨组织,采用免疫组织化学法检测ERK5和MMP-9的表达,并分析ERK5和MMP-9的表达与骨肉瘤患者临床病理特征及预后之间的关系。结果:骨肉瘤组织中ERK5和MMP-9阳性表达率分别为85.9%(61/71)、74.65%(53/71),均明显高于正常骨组织中阳性表达率12.5%(5/40)、10.0%(4/40),差异均具有统计学意义(均P<0.05)。ERK5和MMP-9的阳性表达与Enneking分期和转移相关(均P<0.05)。Kaplan-Meier生存分析显示在骨肉瘤组织中ERK5、MMP-9的阳性表达组患者生存时间均短于阴性表达组(均P<0.05)。单因素Cox回归分析发现,肿瘤大小、Enneking分期、转移、ERK5和MMP-9阳性表达与患者总生存时间相关(均P<0.05);多因素分析证实Enneking分期、转移、ERK5和MMP-9蛋白的阳性表达可作为骨肉瘤患者独立的预后因子(均P<0.05)。结论:ERK5和MMP-9在骨肉瘤组织中高表达,与骨肉瘤的病理特征及预后相关;二者可能在骨肉瘤发生发展的过程中起着重要作用。

IMP3蛋白在骨肉瘤组织中的表达与临床意义

目的:探讨IMP3蛋白在骨肉瘤组织中的表达及其临床意义。方法:选取骨肉瘤患者68例,以骨软骨瘤患者术后石蜡切片20例作为对照,应用SP免疫组化方法检测IMP3蛋白的表达,以分析其在骨肉瘤、骨软骨瘤组织中阳性表达的差异性及与预后的关系。结果:骨肉瘤组织中IMP3阴性、弱、中、强的表达依次为4.41%(3/68)、22.06%(15/68)、22.74%(19/68)、45.59%(31/68),与骨软骨瘤组织相比,IMP3在骨肉瘤的表达存在显著上调(P<0.01)。骨肉瘤中IMP3表达与骨肉瘤患者术后3年存活率呈负相关(P<0.01)。结论:IMP3与骨肉瘤的发生、发展及预后有关,可作为新的骨肉瘤标记物,具有重要的临床意义。

骨肉瘤组织蛋白酶D表达及其与预后的关系

目的：探讨骨肉瘤组织蛋白酶Ｄ表达与肿瘤细胞增殖活性及骨肉瘤预后的关系。方法：运用免疫组织化学染色，检测８５例骨肉瘤细胞增核抗原（ＰＣＮＡ）及组织蛋白酶Ｄ（Ｃａｔｈ－Ｄ）的表达，计算其标记指数（Ｌａｂｅｌｉｎｇｉｎｄｅｘ即ＬＩ）。结果：Ｃａｔｈ－ＤＬＩ在６％～９８．５％之间，ＰＣＮＡＬＩ在９％～９８％之间，Ｃａｔｈ－ＤＬＩ与骨肉瘤患者的年龄、性别、肿瘤的大小及Ｄａｈｉｌｉｎ分型无关，而与ＷＨＯ１９９３年新分型，Ｐｒｉｃｅ分级及ＰＣＮＡＬＩ密切相关。生存率分析显示Ｃａｔｈ－ＤＬＩ＜６２．５％的骨肉瘤预后较好，＞６２．５％的骨肉瘤预后较差。结论：骨肉瘤Ｃａｔｈ－Ｄ表达与肿瘤细胞增殖活性及生存率密切相关。

肠道淋巴瘤临床病理、免疫表型及与EB病毒相关性的研究

目的 研究肠道非霍奇金淋巴瘤的临床病理、免疫表型及与EBV感染的相关性。方法 应用多种抗体标记B淋巴细胞 (CD2 0 ,CD79a ,CD4 5RA ,κ ,λ ,CD5 ,CD10 ,bcl 2 ,cyclinD1)和T淋巴细胞 (CD3ε,CD8,CD4 5RO ,CD5 6 ,GramB ,TIA) ,同时进行细胞增殖活性的检测 (Ki 6 7)、免疫组织化学染色 (SABC法 )、EBV寡核苷酸探针 (EBER)原位杂交。结果 6 0例中 ,免疫组化证实B细胞性淋巴瘤 4 8例 (80 % ) ,T细胞性淋巴瘤 12例 (2 0 % )。EBV EBER原位杂交 4 0例中 6例阳性表达 ,均为T细胞性淋巴瘤 ,阳性细胞占肿瘤细胞的 30 %～ 80 % ,34例B细胞性淋巴瘤EB病毒检测阴性。结论 肠道淋巴瘤以B细胞淋巴瘤多发 ,并以惰性的黏膜相关性边缘区B细胞性淋巴瘤多发 ,与EBV无相关性。肠道T细胞性淋巴瘤侵袭性较强 ,且与EBV相关性较高。