• Genomics & Informatics
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• 제12권4호
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• pp.247-253
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• 2014

Osteosarcoma is the most common primary bone tumor, generally affecting young people. While the etiology of osteosarcoma has been largely unknown, recent studies have suggested that cyclooxygenase-2 (COX-2) plays a critical role in the proliferation, migration, and invasion of osteosarcoma cells. To understand the mechanism of action of COX-2 in the pathogenesis of osteosarcoma, we compared gene expression patterns between three stable COX-2-overexpressing cell lines and three control cell lines derived from U2OS human osteosarcoma cells. The data showed that 56 genes were upregulated, whereas 20 genes were downregulated, in COX-2-overexpressed cell lines, with an average fold-change > 1.5. Among the upregulated genes, COL1A1, COL5A2, FBN1, HOXD10, RUNX2, and TRAPPC2 are involved in bone and skeletal system development, while DDR2, RAC2, RUNX2, and TSPAN31 are involved in the positive regulation of cell proliferation. Among the downregulated genes, HIST1H1D, HIST1H2AI, HIST1H3H, and HIST1H4C are involved in nucleosome assembly and DNA packaging. These results may provide useful information to elucidate the molecular mechanism of the COX-2-mediated malignant phenotype in osteosarcoma.

• Asian Pacific Journal of Cancer Prevention
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• 제14권3호
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• pp.2049-2052
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• 2013

The effects of polymorphisms in ERCC5, ERCC6, XPC, CCNH and MMS19L on osteosarcoma response to chemotherapy and the survival of the affected patients were assessed. Genotyping of ERCC5, ERCC6, XPC, CCNH and MMS19L was performed by PCR-RFLP assay. The median PFS was 12.8 months, and the median OS was 18.6 months. Individuals carrying homozygous genotypes of ERCC5 rs17655 and ERCC5 rs1047768 were more like to have good response to treatment, while those carrying homozygous genotypes of MMS19L rs29001322 showed poor response. Osteosarcoma patients carrying TT genotype of ERCC5 rs1047768 showed a significantly longer PFS (16.8 months) and OS (21.4 months) than CC genotype, with HRs(95% CI) of 0.31 (0.10-0.93) and 0.32 (0.06-0.97), respectively. Conversely, those with the TT genotype of MMS19L rs29001322 demonstrated shorter PFS and OS, the HRs (95% CI) being 2.23 (1.08-4.15) and 4.62 (1.45-16.08), respectively. Our findings showed polymorphisms in ERCC5 rs1047768 and MMS19L rs29001322 to be associated with clinical outcome of osteosarcoma patients undergoing chemotherapy.

• Asian Pacific Journal of Cancer Prevention
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• 제13권6호
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• pp.2847-2851
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• 2012

Purpose: To study the killing effects on osteosarcoma cells of cinobufacini and cisplatin in combination and the related mechanisms so as to explore the chemotherapeutic method with integrated traditional Chinese and Western medicines. Methods: Cinobufacini and cisplatin were applied to OS732 cells singly or jointly and survival rates were measured by MTT assay. Changes in cellular shape were observed with inverted phase contrast and fluorescence microscopy and apoptosis rates were analyzed with flow cytometry (FCM). Immunocytochemistry were used to examine the Fas expression of OS732 cells. Results: The combination of cinobufacini and cisplatin had the effect of up-regulating Fas expression and inducing apoptosis. The survival rate of combined application of 100 ${\mu}g/ml$ cinobufacini and 1 ${\mu}g/ml$ cisplatin on OS-732 cells was significantly lower than with either of the agents alone (p<0.01). Changes in cellular shape and apoptotic rates also indicated the apoptosis-inducing effects of combined application were much enhanced. Conclusion: The combination of cinobufacini and cisplatin demonstrated strong killing effects on OS-732 cells which might be related to up-regulation of Fas expression.

• Molecules and Cells
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• 제41권6호
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• pp.523-531
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• 2018

Tumour metastasis is one of the most serious challenges of cancer as it is the major cause of mortality in patients with solid tumours, including osteosarcoma (OS). In this regard, anti-metastatic genes have potential for metastasis inhibition strategies. Recent evidence showed the importance of breast cancer metastasis suppressor 1 (BRMS1) in control of OS invasiveness, but the regulation of BRMS1 in OS remains largely unknown. Here, we used bioinformatics analyses to predict BRMS1-targeting microRNAs (miRNAs), and the functional binding of miRNAs to BRMS1 mRNA was evaluated using a dual luciferase reporter assay. Among all BRMS1-targeting miRNAs, only miR-151b, miR-7-5p and miR-3200-5p showed significant expression in OS specimens. Specifically, we found that only miR-3200-5p significantly inhibited protein translation of BRMS1 via pairing to the 3'-UTR of the BRMS1 mRNA. Moreover, we detected significantly lower BRMS1 and significantly higher miR-3200-5p in the OS specimens compared to the paired adjacent non-tumour bone tissues. Furthermore, BRMS1 and miR-3200-5p levels were inversely correlated to each other. Low BRMS1 was correlated with metastasis and poor patient survival. In vitro, overexpression of miR-3200-5p significantly decreased BRMS1 levels and promoted OS cell invasion and migration, while depletion of miR-3200-5p significantly increased BRMS1 levels and inhibited OS cell invasion and migration. Thus, our study revealed that miR-3200-5p may be a critical regulator of OS cell invasiveness.

• Asian Pacific Journal of Cancer Prevention
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• 제14권12호
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• pp.7081-7084
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• 2013

MicroRNAs have been demonstrated to regulate proliferation and apoptosis in many types of cancers, but biological functions in osteosarcomas remain relatively unknown. Here, we found expression of miR-802 to be up-regulated in osteosarcoma tissues in comparison with adjacent normal tissues. Enforced expression of miR-802 was able to promote cell proliferation in U2OS and MG63 cells, while miR-802 antisense oligonucleotides (antisense miR-802) inhibited cell proliferation. At the molecular level, our results further revealed that expression of p27, a negative cell-cycle regulator, was negatively regulated by miR-802. Therefore, the data reported here indicate that miR-802 is an important regulator in osteosarcoma, our findings contributing to a better understanding of important mis-regulated miRNAs in this tumour type.

• 대한골관절종양학회지
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• 제17권1호
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• pp.23-29
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• 2011

목적: 본 연구에서는 골육종 세포내 PTEN 발현정도가 세포 성장과 트로글리타존에 대한 반응도에 미치는 영향에 대해 알아보고자 하였다. 대상 및 방법: 웨스턴 블롯 분석을 통해 트로글리타존 처리 후 PTEN 발현 정도를 관찰하였고, WST를 통해 세포 증식정도를 측정하였다. 야생형 PTEN 및 돌연변이형 PTEN 발현시키는 플라스미드 DNA를 트랜스펙션하여 PTEN 발현 정도를 측정하였다. 결과: 사람골육종 세포주 U-2OS는 트로글리타존 처리 농도 및 시간에 비례하여 증식 억제를 보였고, 세포내 PTEN 발현 정도는 트로글리타존 처리 농도에 비례하여 증가하였다. 트로글리타존을 이용하여 U-2OS세포 내 PTEN 발현을 증가시키면 세포 성장 억제와 세포사 유도가 나타났다. 또한 플라스미드 트랜스펙션에 의한 PTEN 과발현은 트로글리타존의 세포증식 억제 효과를 증가 시키며 돌연변이형 PTEN을 트랜스펙션 시키는 경우 세포증식효과는 관찰되지 않았다. 결론: 골육종 세포내 PTEN 과발현이 트로글리타존에 의한 골육종 세포의 증식 억제 및 세포사 유도와 관련 되어있음을 알 수 있으며, 세포내에 PTEN이 과발현 된 상태에서 트로글리타존의 효과가 증가됨을 알 수 있었다.

• Asian Pacific Journal of Cancer Prevention
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• 제16권15호
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• pp.6451-6455
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• 2015

Background: Primary bone neoplasms are rare, contributing only 0.2% of the global burden of all human malignancies. Osteosarcoma (OS) and chondrosarcoma (CS) are the most common malignancies of bone. The giant cell tumor of bone (GCTb) is a benign tumor with behavior characterized by osteolytic bone destruction. The OS, CS and GCTb affect both sexes, all races and generally have incidence peaks regarding the age of the patient which vary according to the tumor type. We analyzed the incidences of OS, CS and GCTb and their relations with gender and age in patients treated in the National Rehabilitation Institute (INR, for its acronym in Spanish) over a period of nine years. Materials and Methods: In the study period, clinic pathological data for 384 patients were obtained with clinical, radiological and histopathological diagnosis for OS, GCTb and CS. Data analysis was performed using the chi-square and Fisher's exact tests. Results: From 2006 to 2014 were recorded 384 cases of bone malignancies in the database of INR. The GCTb had the highest incidence (53.1%), followed by OS (31.3%) and finally the CS (15.6%). The overall average age was $33.6{\pm}15.8$ years and the overall frequency of gender had a ratio of 1/1.03 male/female. The states with the highest incidence were Distrito Federal and Estado de Mexico with 29.2% and 25.3% respectively. Malignant neoplasms of bone assessed in the course of nine years show three significant increases in 2008, 2011 and 2014 (p=0.14). We found association between sex and tumor type (p=0.03), GCTb and CS predominated in females (54.9% and 56.6% respectively), while for the OS males were most affected (59.1%). Age was different in relation with tumor type (p=0.0001), average age was $24.3{\pm}11.2$ years for OS, $34.5{\pm}13$ years for GCTb and $49.2{\pm}18.5$ years for CS. Furthermore, associations of tumor type with topographic location of the primary tumor (P=0.0001) were found. Conclusions: In this study we can see that incidence of musculoskeletal tumor in our population is continuously increasing and in nine years an approximately 200% increase of musculoskeletal tumor cases was observed.

• 대한골관절종양학회지
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• 제15권2호
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• pp.111-121
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• 2009

배경: 골육종은 소아 및 청소년기에 가장 흔하게 발생하는 악성 골종양 중 하나이다. 최근 수술적 치료와 항암 화학요법을 병행하여 생존률이 증가하였지만, 아직까지도 항암제는 항암제 내성 및 이차성 악성 종양의 발생 등 여러 문제점을 가지고 있다. 일부 종양은 matrix metalloproteinase(MMPs)의 발현이 증가되어 있고, MMP inhibitor에 대한 연구가 진행되고 있다. 반면 bisphosphonate(BPs) 제제는 골흡수를 억제하는 능력이 있으며, 파골세포와 관계된 골 병변에 광범위하게 사용되고 있다. 또 bisphosphonate 제제는 직접적인 항암효과를 가지고 있는 것으로 알려져 있다. 대상 및 방법: 골육종 세포주(U2OS)를 ibandronate(0, 0.1, 1, 10M)를 이용하여 48시간 동안 처치하였다. 세포의 생존능은 MTT assay를 이용하여, MMP-2와 MT1-MMP의 mRNA level은 reverse-transciption polymerase chain reaction을 이용하여 측정하였으며, MMP-2와 MT1-MMP 단백의 양은 Westernblot을, MMP-2의 활성은 Gelatin zymography를 이용하여 측정하였다. 또, ibandronate 처치 전후의 골육종 세포주의 침습성은 Matrigel invasion assay를 이용하여 측정하였다. 결과: 48시간 ibandronate 처치 후 U2OS 세포주의 침습력은 ibandronate에 대해 용량 의존적으로 감소하였다. 특히 10M 이내의 ibandronate는 세포독성이 나타나지 않았다. 젤라틴 융해능과 MMP-2 및 MT1-MMP의 단백 및 mRNA 정도역시 ibandronate 농도가 증가할수록 감소하였다. 결론: Ibandronate는 골육종 세포주의 MMP-2 및 MT1-MMP의 발현을 억제하였으며, 종양세포의 침습력을 감소시켰다. Bisphosphate의 종양세포 침습 억제 능력은 새로운 전이 억제제의 개발에 도움이 될 것으로 사료된다.

• Asian Pacific Journal of Cancer Prevention
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• 제16권17호
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• pp.7689-7694
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• 2015

Background: Bone tumors are neoplasias with a high overall mortality; one of the main factors that reduce survival is their high capacity to develop metastases. It has been reported that finding lung metastases at diagnosis of osteosarcoma (OS), chondrosarcoma (CS) and giant cell tumor of bone (GCTb) is quite common. In this study, we inquire the relationship of metastases caused by these tumors with different clinical and pathological aspects, in order to guide medical personnel in the diagnosis and opportune treatment of metastases or micro metastases. Materials and Methods: We collected data of 384 patients with clinical, radiological and histopathological diagnosis of OS, GCTb and CS that attended the National Rehabilitation Institute (INR) during 2006 to 2014. Chi-square and Fisher's exact tests were performed for data analysis. Results: In the three tumor types, the presence of metastases at diagnosis was variable (p=0.0001). Frequency of metastases was 36.7%, 31.7% and 13.2% for OS, CS and GCTb respectively. The average age had no significant difference (p>0.05) in relation to metastases, even so, patients with OS and GCTb and metastases, were older while patients with CS and metastases were younger, in comparison to patients without metastases. Males had a higher frequency of metastases (68.2%, p = 0.09) in contrast to CS and GCTb, in which the metastases was more frequent in women with 51.9% (p = 0.44) and 57.9% (p = 0.56) respectively. Broadly, metastasis was associated with primary tumors located in the femur (44.4%), followed by the tibia (15.6%); metastases was more frequent when primary tumor of GCTb and OS were in the same bones, but were located in the hip (26.3%) for CS. Conclusions: The frequency of metastases in OS, GCTb and CS is high in our population and is determined by different clinicopathological variables related to the kind of tumor. Further studies are needed in order to evaluate metastases subsequent to diagnosis and associations with survival and clinicopathological factors, as well as to determine the sensitivity and specificity of current methods of detection.

• Clinical and Experimental Pediatrics
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• 제59권9호
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• pp.374-380
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• 2016

Purpose: Patients with unresectable, relapsed, or refractory osteosarcoma need a novel therapeutic agent. Metformin is a biguanide derivative used in the treatment of type II diabetes, and is recently gaining attention in cancer research. Methods: We evaluated the effect of metformin against human osteosarcoma. Four osteosarcoma cell lines (KHOS/NP, HOS, MG-63, U-2 OS) were treated with metformin and cell proliferation was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Cell cycle progression and apoptosis were evaluated using flow cytometric analysis, and migration and wound healing assay were performed. Fourteen female Balb/c-nude mice received KHOS/NP cell grafts in their thigh, and were allowed access to metformin containing water (2 mg/mL) ad libitum. Tumor volume was measured every 3-4 days for a period of 4 weeks. Results: Metformin had a significant antiproliferative effect on human osteosarcoma cells. In particular, metformin inhibited the proliferation and migration of KHOS/NP cells by activation of AMP-activated protein kinase and consequent inhibition of the mammalian target of rapamycin pathway. It also inhibited the proliferation of cisplatin-resistant KHOS/NP clone cells. Analysis of KHOS/NP xenograft Balb/c-nude models indicated that metformin displayed potent in vivo antitumor effects. Conclusion: Further studies are necessary to explore metformin's therapeutic potential and the possibilities for its use as an adjuvant agent for osteosarcoma.