골육종조직에서 활성화된 Neutral RNase Isozyme의 정제와 작용기전에 관한 연구

최충혁,최일용,김성준,고재경 한양대학교 의과대학 1994 한양의대 학술지 Vol.14 No.2

Of the RNase isozymes isolated from osteosarcoma tissue of human, ribonuclease(RNase) isozyme I exhibited the highest activity and appeared to be activated. The present study was carried out to purify the RNase isozyme I and to determine the substrate specificity for the isozyme specfic to osteosarcoma. Activities of both RNase and RNase inhibitor in osteosarcoma tissue were markedly increased, indicating the possible use of the enzyme and the enzyme inhibitor as biochemical marker for osteosarcoma. RNases in osteosarcoma tissue were separated by a DEAE-cellulose column chromatography into five isozymes, of which two isozymes were activated and other two isozymes were specific to osteosarcoma. The activity of RNase inhibitor complexed with these RNase isozymes was observed to be high. Of the two RNase isozymes activated, RNase isozyme I exhibited higher activity and appeared to be specific to osteosarcoma. The RNase isozyme I was separated by a HPLC into 7 subpeaks, of which RNase isozyme I-3 with the highest RNase activity appeared to by specific to osteosarcoma. The RNase isozyme I-3 separated and purified from osteosarcoma tissue was not active against double stranded polynucleotides, but active against single stranded polyribonucleotide, being highly active against single stranded polyribonucleotide, being highly active on C-C, C-U and A-U linkages. A considerable activity was observed with RNA as substrate, being different from other RNase isozymes isolated from osteosarcoma tissue and its control bone tissue. The results obtained in the present study indicated that the RNase isozyme I-3 results obtained in the present study indicated that the RNase isozyme I-3 purified from osteosarcoma tissue was activated, specific to osteosarcoma and exhibited a considerable acitivty on RNA, suggesting that the isozyme play an important role in RNA mediated tumorigenesis in osteosarcoma.

Zoledronic acid가 골육종 세포에서 Matrix Metalloproteinase-2와 -14의 발현과 폐전이에 미치는 영향

김용백,김상범,권영호,김재도 KOSIN UNIVERSITY COLLEGE OF MEDICINE 2006 高神大學校 醫學部 論文集 Vol.21 No.1

Purpose : Gelatinase A (MMP-2) and membrane-type metalloproteinase (MT1-MMP, MMP-14) play a critical role in metastasis. It was known that bisphosphonate acts not only to protect bone absorption but also to inhibit tumor invasion and metastasis. The aim of this study is to investigate the inhibition of MMP-2 and 14 in osteosarcoma by zoledronic acid, whether it can prevent lung metastasis in nude mouse. Material and Method : Protein synthesis and mRNA expression of MMP-2 and MMP-14 cultured human MG-63 osteosarcoma and HOS osteosarcoma cell lines with zoledronic acids were measured. In vivo test, it was observed that the inhibitory effect of zoledronic acid on lung metastasis of osteosarcoma with nude mouse model. Result : Zoledronic acid inhibited mRNA expression of MMP-14 and MMP-2 in cultured human MG-63 osteosarcoma and HOS osteosarcoma cell lines. Zoledronic acid decreased the intensity of activated 62KD MMP-2 in human MG-63 osteosarcoma and HOS osteosarcoma cell lines on gelatine zymography. Number of metastatic lung nodule decreased from 9.4 to 1.2 in MG-63 osteosarcoma cell line and from 8 to 1.6 in HOS osteosarcoma cell line with nude mouse model. Conclusion : In vitro, zoledronic acid had an inhibitory effect on the MMP-14 and MMp-2, which also prevents lung metastasis of osteosarcoma in vivo. So, zoledronic acid could be used as one of good chemotherapeutic agents against metastasis of osteosarcoma.

Zoledronic acid가 골육종 세포에서 Matrix Metalloproteinase-2와 44의 발현과 폐전이에 미치는 영향

김용백,김상범,권영호,김재도 고신대학교(의대) 고신대학교 의과대학 학술지 2006 고신대학교 의과대학 학술지 Vol.21 No.1

Purpose : Gelatinase A (MMP-2) and membrane-type metalloproteinase (MT1-MMP, MMP-14) play a critical role in metastasis. It was known that bisphosphonate acts not only to protect bone absorption but also to inhibit tumor invasion and metastasis. The aim of this study is to investigate the inhibition of MMP-2 and 14 in osteosarcoma by zoledronic acid, whether it can prevent lung metastasis in nude mouse. Material and Method : Protein synthesis and mRNA expression of MMP-2 and MMP-14 cultured human MG-63 osteosarcoma and HOS osteosarcoma cell lines with zoledronic acids were measured, In vivo test, it was observed that the inhibitory effect of zoledronic acid on lung metastasis of osteosarcoma with nude mouse model Result : Zoledronic acid inhibited mRNA expression of MMP-14 and MMP-2 in cultured human MG-63 osteosarcoma and HOS osteosarcoma cell lines. Zoledronic acid decreased the intensity of activated 62KD MMP-2 in human MG-63 osteosarcoma and HOS osteosarcoma cell lines on gelatine zymography. Number of metastatic lung nodule decreased from 9.4 to 1.2 in MG-63 osteosarcoma cell line and from 8 to 1.6 in HOS osteosarcoma cell line with nude mouse model. Conclusion : In vitro, zoledronic acid had an inhibitory effect on the MMP-14 and MMP-2, which also prevents lung metastasis of osteosarcoma in vivo. So, zoledronic acid could be used as one of good chemotherapeutic agents against metastasis of osteosarcoma.

Interactive Effect of Bisphenol A (BPA) Exposure with -22G/C Polymorphism in LOX Gene on the Risk of Osteosarcoma

Jia, Jie,Tian, Qing,Liu, Yong,Shao, Zeng-Wu,Yang, Shu-Hua Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.6

Background: Osteosarcomas have many established risk factors, both genetic and environmental, but by themselves these explain only part of the total cancer incidence. Bisphenol A (BPA) is an environmental estrogen associated with risk of several kinds of tumour. The lysyl oxidase gene (LOX) may also contribute to risk of tumours including osteosarcomas. Here, we investigated possible interactions of BPA and a LOX polymorphism on the risk of osteosarcoma. Method: The present hospital-based case-control study included 106 cancer patients and 112 controls from a Chinese population. Internal burden of BPA exposure was assessed using high-performance liquid chromatography-mass spectrometry (HPLC-MS) method. Genotypes were determined using PCR-RFLP methods. Results: Compared with those in low BPA exposure group, subjects with BPA more than or equal to median value had significant increased risk of osteosarcoma among subjects who carried GC or CC genotypes. A significant interaction with BPA level and the -22G/C polymorphism was observed for osteosarcoma overall, osteosarcoma affecting knee and osteosarcoma affecting hip, as $P_{forinteraction}$ = 0.036 for osteosarcoma overall; $P_{forinteraction}$ = 0.024 for osteosarcoma affecting knee; and $P_{forinteraction}$ = 0.017 for osteosarcoma affecting hip. Conclusions: The results suggest that BPA exposure interacts with the -22G/C polymorphism of the LOX gene to increase the risk of osteosarcoma.

Radiographic manifestations of fibroblastic osteosarcoma: A diagnostic challenge

Tahmasbi-Arashlow, Mehrnaz,Barnts, Kelcie Louise,Nair, Madhu K.,Cheng, Yi-Shing Lisa,Reddy, Likith V. Korean Academy of Oral and Maxillofacial Radiology 2019 Imaging Science in Dentistry Vol.49 No.3

Osteosarcoma is the most common primary bone tumor after plasma cell neoplasms. Osteosarcoma has diverse histological features and is characterized by the presence of malignant spindle cells and pluripotent neoplastic mesenchymal cells that produce immature bone, cartilage, and fibrous tissue. Osteosarcoma most frequently develops in the extremities of long bones, but can occur in the jaw in rare cases. The clinical and biological behavior of osteosarcoma of the jaw slightly differs from that of long-bone osteosarcoma. The incidence of jaw osteosarcoma is greater in the third to fourth decades of life, whereas long-bone osteosarcoma mostly occurs in the second decade of life. Osteosarcoma of the jaw has a lower tendency to metastasize and a better prognosis than long-bone osteosarcoma. Radiographically, osteosarcoma can present as a poorly-defined lytic, sclerotic, or mixed-density lesion with periosteal bone reaction response. Multi-detector computed tomography is useful for identifying the extent of bone destruction, as well as soft tissue involvement of the lesion. The current case report presents a fibroblastic osteosarcoma involving the left hemimandible with very unusual radiographic features.

Radiographic manifestations of fibroblastic osteosarcoma: A diagnostic challenge

Mehrnaz Tahmasbi-Arashlow,Kelcie Louise Barnts,Madhu K. Nair,Yi-Shing Lisa Cheng,Likith V. Reddy 대한영상치의학회 2019 Imaging Science in Dentistry Vol.49 No.3

Osteosarcoma is the most common primary bone tumor after plasma cell neoplasms. Osteosarcoma has diverse histological features and is characterized by the presence of malignant spindle cells and pluripotent neoplastic mesenchymal cells that produce immature bone, cartilage, and fibrous tissue. Osteosarcoma most frequently develops in the extremities of long bones, but can occur in the jaw in rare cases. The clinical and biological behavior of osteosarcoma of the jaw slightly differs from that of long-bone osteosarcoma. The incidence of jaw osteosarcoma is greater in the third to fourth decades of life, whereas long-bone osteosarcoma mostly occurs in the second decade of life. Osteosarcoma of the jaw has a lower tendency to metastasize and a better prognosis than long-bone osteosarcoma. Radiographically, osteosarcoma can present as a poorly-defined lytic, sclerotic, or mixed-density lesion with periosteal bone reaction response. Multi-detector computed tomography is useful for identifying the extent of bone destruction, as well as soft tissue involvement of the lesion. The current case report presents a fibroblastic osteosarcoma involving the left hemimandible with very unusual radiographic features.

골육종의 억제과정에 미치는 Ribonuclease 의 작용

김성준,이경태,고재경 한양대학교 의과대학 1993 한양의대 학술지 Vol.13 No.1

Neutral ribonuclease (RNase) highly active toward polycyidylate (poly C) was isolated from osteosarcoma tissue by DEAE-cellulose column chromatography and was found to be specific to the bone sarcoma by means of a high performance liquid chromatography (HPLC) and polyacrylamide gel electrophoresis (PAGE). Also studied were substrate specificity and the product analysis of the neutral RNase to understand the role of the enzyme specific to osteosarcoma in pathogenesis of the bone sarcoma. Neutral RNase activity was greatly increased in osteosarcoma tissue, suggesting that in could by used as a marker for the bone sarcoma. Neutral RNases in osteosarcoma tissue were separated by a DEAE-cellulose column chromatography into 5 peaks, of which peaks Ⅵ and Ⅶ RNase isozymes were not found in the control bone tissue. The activity of peak Ⅰ RNase isozyme was most active among the RNase isozymes separated. HPLC and PAGE patterns for the proteins associated with the peak Ⅰ neutral RNase of osteosarcoma appeared to be different those of the control, suggesting that the enzyme might be specific to the bone sarcoma. The peak Ⅰ neutral RNase isozyme specific to the osteosarcoma was found to be active toward ss polyribonucleotide (highly active toward C-C, C-U and A-U linkages). Majority of the poly C digest by the RNase specific to the osteosarcoma was observed to be oligoribonucleotides with chain length of 4-25, indicating that the enzyme was endonuclease in nature. The present study indicate that 1) neutral RNase activity was greatly increased in osteosarcoma tissue, 2) the enzyme could be used as a marker for osteosarcoma, 3) the peak Ⅰ neutral RNase isozyme isolated from osteosarcoma tissue might be specific to the bone sarcoma, 4) the RNase isozyme was active toward ss polyribonucleotide and 5) the RNase isozyme was endofibonuclease in nature, suggesting a possible role of the RNase isozyme in the suppression of the osteosarcoma.

Correlation of IGF1R expression with ABCG2 and CD44 expressions in human osteosarcoma

김충규,오선주,김숙자,임선희,허정훈,정소학 한국유전학회 2018 Genes & Genomics Vol.40 No.4

Osteosarcoma is the most common type of malignant bone tumors. Insulin Growth Factor 1 receptor (IGFR1) has been known as a prognostic factor for metastasis of osteosarcoma. ABC subfamily G member2 (ABCG2) is related to resistance to anti-cancer drug, and CD44 has a role in tumor growth and metastasis. The purpose of this study is to investigate the relationship among expression patterns of IGF1R, ABCG2, and CD44 in osteosarcoma. The expression levels of IGF1R, ABCG2, and CD44 proteins were determined in tissue arrays containing osteosarcoma tissues from 59 osteosarcoma patients. The expression pattern of IGF1R was highly correlated with the expression pattern of ABCG2 (r = 0.88) in overall osteosarcoma patients. According to pathological types, the expression pattern of IGF1R showed the higher correlation with ABGC2 (r = 0.90) and CD44 (r = 0.61) in osteoblatic type than in chondroblastic type. According to gender with pathologic type, the correlation between the expression patterns of IGF1R and CD44 was higher in male with osteoblatic type than in female with osteoblatic type. Among different age groups, the 1–10 years age group showed higher correlation in IGF1R versus CD44 (r = 0.90) and ABCG2 versus CD44 (0.80) than in other age groups. These results showed that the expression of IGF1R appears to be highly correlated with the expression of ABCG2 in osteosarcoma and with the expression of CD44 in osteosarcoma patients under age of 10, which suggests that ABCG2 and CD44 can be used as prognostic factors with IGF1R for specific prognosis and efficient treatment of osteosarcoma.

Epidermal Growth Factor Receptor: Is It a Feasible Target for the Treatment of Osteosarcoma?

이준아,고윤미,김동호,임중섭,Chang-Bae Kong,Wan Hyeong Cho,Dae-Geun Jeon,Soo-Yong Lee,고재수 대한암학회 2012 Cancer Research and Treatment Vol.44 No.3

Purpose Features of epidermal growth factor receptor (EGFR) expression in osteosarcoma and in vitro efficacies of EGFR inhibitors against osteosarcoma cells were evaluated. Materials and Methods Thirty biopsy samples of osteosarcoma patients were retrospectively analyzed for EGFR protein expression by immunohistochemistry. Relationships between EGFR expression and clinicopathologic characteristics and treatment outcomes were evaluated. Four osteosarcoma cell lines were analyzed for EGFR and p-EGFR expression by western blotting. Efficacies of gefitinib and BIBW2992 on osteosarcoma cells were evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Tyrosine kinase domains in exons 18 to 21 were sequenced and gene expression analyses of EGFR and PTENwere performed in four osteosarcoma cell lines. Results EGFR protein was expressed in 27 (90%) samples (6 low, 12 intermediate, 9 high) and in three cell lines. Intermediate or high staining for EGFR was related to a tumor volume<150 mL (p<0.001)and histologic subtype other than osteoblastic type (p=0.03). However, EGFR expression was not associated with histologic response to preoperative chemotherapy or survival. Gefitinib and BIBW 2992 did not have any significant inhibitory effect on cell viabilities. DNA sequencing analysis revealed three osteosarcoma cell lines have single base changes at codon 2361 of exon 20 (G to A), without affecting translation results. Furthermore, no mutation was found to be associated with constitutive EGFR activation. Conclusion In the present study, gefitinib and BIBW2992 were not effective against osteosarcoma cells. However, as osteosarcoma cells express EGFR, further studies are necessary to explore the potential of other therapeutic agents targeting EGFR.

PIK3CA and AKT Gene Polymorphisms in Susceptibility to Osteosarcoma in a Chinese Population

He, Mao-Lin,Wu, Yang,Zhao, Jin-Min,Wang, Zhe,Chen, Ying-Bin Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.9

Purpose: To explore the association between PIK3CA and AKT single nucleotide polymorphisms(SNP) and osteosarcoma susceptibility. Methods: TaqMan polymerase chain reaction(PCR) was used to detect the genotypes of SNPs (rs7646409, rs6973569 and rs9866361) in peripheral blood samples from 59 patients with osteosarcoma and from 63 healthy controls. Unconditional logistic regression was used to analyze the correlation between SNPs and osteosarcoma risk. Results: No statistically significant difference was found between osteosarcoma patients and healthy controls in the genotype of AKT rs6973569 (P=0.7). However, after stratified analysis, the genotype AA of AKT rs6973569 carried a higher risk of osteosarcoma metastasis (OR:2.94, 95%CL:1.00-8.59); the difference of rs7646409 genotype distributions between the case and control groups was statistically significant (P=0.032). Taking genotype TT as a reference, the risk of osteosarcoma increased three fold in patients with genotype CC (OR:3.47, 95%CL:1.26-9.56). A statistically significant difference was found between the alleles C and T (P=0.005). Further analysis showed that the risk factor was more pronounced in male patients with Enneking's stage IIB and osteoblastic osteosarcoma. PIK3CA rs9866361 did not fit Hardy-Weinberg equilibrium (P<0.05). Conclusions: Genotype CC in locus PIK3CA rs7646409 may increase the risk of osteosarcoma in the Chinese population.