사람 치은모세포에서 다양한 세포외기질 분자에 light-emitting diode 조사 효과

박형근,장윤영,정권,고미경,장미선,국은별,정민아,임원봉,임채광,김옥준,최홍란 대한구강악안면병리학회 2006 대한구강악안면병리학회지 Vol.30 No.3

It has been reported that light-emitting diodes(LED) can be used in the treatment of oral diseases. Although bio-stimulatory effects of LED irradiation such as promotion of wound healing have been well known, there are few reports about molecular mechanisms associated with wound healing by LED irradiation. The purpose of the present study was to investigate the expression pattern of various extracellular matrix(ECM) molecules in relation to wound healing after LED irradiation on primary human gingival fibroblasts(hGFs) in vitro. The source of light for irradiation was a continuous-wave LED emitting at a wavelength of 635 nm, and manufactured that energy density was 5 mW/cm2 on sample surfaces. The hGFs were irradiated for 1 hour at 37℃ in 5% CO2 humidified chamber. Experimental samples were acquired at 0 (right after irradiation), 24 and 48 hour after irradiation. To investigate the molecular mechanisms associated with wound healing, we examined the mRNA expression of 6 types of collagens, 7 types of matrix metalloproteinases(MMPs) and 4 types of tissue inhibition of metalloproteinases(TIMPs) after LED irradiation by RT-PCR. The mRNA expression of collagen 4, MMP-3, 9, and 16, and TIMP-3 was influenced by LED irradiation. Generally, the collagen expression of the irradiation group was slightly increased, particularly collagen 4 was significantly increased at 0 hour. The expression of MMP-3 was increased at 0 and 24 hours and MMP-16 was increased at 24 hours, respectively. The expression of MMP-9 was decreased at 0 hour and increased at 24 and 48 hours. The mRNA expression of TIMP-3 was significantly decreased at 24 and 48 hours after irradiation. These results suggest that the altered expression of ECM molecules after LED irradiation may contribute to the accelerated wound healing.

백서에서 창상 치유에 대한 Líght Emitting Diode(LED) 조사의 효과

김용찬,고미경,장미선,국은별,정권,장윤영,정민아,양재혁,임채광,임원봉,박진수,김옥준,최홍란 대한구강악안면병리학회 2005 대한구강악안면병리학회지 Vol.29 No.3

LEDs have been shown to be a safe, efficient, light-weight, and less-expensive alternative to heal wound. LED irradiation at the same biostimulatory wavelength of previous laser studies have similar biochemical effects The purpose of present study is to evaluate the effects of wound healing by LED irradiation. Thirty 34-day-old sprague dawley were used for present study. 1.5mm diameter defected holes were formed in both ear lobes of rat by rubber dam punch. 635nm and 890nm irradiation was performed by LED for 2 weeks, followed by histologíc examination staíned with H&E and Masson trichrome. Also, RT-PCR was carried out to find out the mRNA expression level in gingival fjbroblast irradiated by 635nm for 1 hour. In gross exarnination, wound healing was observed in irradiated group comparing to control For microscopic exarnination, repair by connective tissues was filled in defects of irradiated group, while dense cellular bands consisting of fjbroblasts and capi llaries were found at the end of defect in control By staining of masson trichrome, amount of collagens were found in irradiated group. In a result of RT-PCR, mRNA expressions of TGF- ß , MMP-1,3 and Timp-3 were down-regulated in irradiated group comparing with their expression in control group. Taken together, LED irradiation increase the prolifeation and the activity of fibroblasts and down-regualted the TGF-ß , MMP- 1,3 and Timp- 3 mRNA, followed by activation of would healing.

4-NQO를 이용한 백서의 구강암 발암과정에서 나타나는 유전자의 변화

김대업,장윤영,정권,고미경,장미선,국은별,임원봉,임채광,양재혁,황윤찬,황인남,오원만,김옥준,최홍란 대한구강악안면병리학회 2005 대한구강악안면병리학회지 Vol.29 No.5

The number 01' patien ts with tongue carcinoma is increasing rapidly among young indiv idua ls in many parts of the worl d. Until now‘ most of studies were focused on the comparison malignancy wi th normal 01' dysplasia. There is little report of gene a lterations in normal to cancer , oral carcinogenesis. The purpose of present study is to evaluate the gene a ltc ration in every steps of oral carcinogenes is by DD- PCR. To induce tongue carcinoma in rat by 4- NQO. each dri nking water made 10ppm. 25ppm. 50ppm and control(o nly D.W without 4-NQO) . Specimens were classified into 4 groups s uch as co ntrol, I(mild & mocle rate dysplasia) , II(severe dysplasia and car cinoma in s itu) , III(carcinoma). Total RNA was ext racted and DD- PCR was performed using customized random primers. And to confïrmed t he results of DD-PCR‘ RT- PCR a nd real-time PCR with specific primers were carried out. There was phenotypic alteration in tongue 。f dosc a nd t imc dc pcndcnt man ncr. In gross examination, multiple papules, patch form or ulcerations were observed during 4 - NQO t reatment Hi s tologicall y, dysplasia was observed in 3 to 6 month and tumor formation in 6 to 8 month For DD-PCR, RT-PCR and real-time PCR, cyclophilin A, BAC RP23-372MB and BAC CH230-103E9 were differ entia lly expressed. Taken together, cyclophilin A has a role in all steps of oral carcinogenesis. BAC RP23-372N田is implica ted in carcinoma in s itu a nd BAC CH230-103E9 mRNA expression is assoicated with dysplasia and carcinom in s itu Conclus ively. some genes a re impli catcd a ll st eps of oral carci nogenesis, others are associated with one step, whi ch meant that genes are di fferentia lly expressed in every steps

백서 혀에서의 4-nitroquinoline 1-oxide 유도 발암과정에서 Bcl-2 계 유전자의 발현

최재욱,정성수,이금숙,김병국,김재형,국은별,장미선,고미경,정권,최홍란,김옥준 대한안면통증∙구강내과학회 2005 Journal of Oral Medicine and Pain Vol.30 No.3

The number of patients with tongue carcinoma is increasing rapidly among young individuals in many parts of the world. Oral carcinoma progresses from hyperplastic lesion through dysplasia to invasive carcinoma and the concept of "field cancerization" with molecular alteration has been suggested for oral cavity carcinogenesis. Significant improvement in treatment and prognosis will depend on more detailed understanding of the multi-step process leading to cancer development. To induce tongue carcinoma in rat by 4-NQO, each drinking water was made to 10 ppm, 25 ppm, 50 ppm and control (only D.W. without 4-NQO). Specimens were classified into 4 groups such as control, I (mild & moderate dysplasia), II (severe dysplasia and carcinoma in situ), III (carcinoma). The mRNA expressions of Bcl-2 family were evaluated by RT-PCR technique. For anti-apoptotic Bcl-2 family, mRNA expression of Bcl-w was down-regulated in all stages of tongue carcinogenesis model. However, mRNA expression of Bcl-2 was up-regulated. For pro-apoptotic Bcl-2 family, all members were down-regulated in all stages of tongue carcinogenesis model except for Bad mRNA in group III. In terms of BH3 only protein, mRNA expressions of Bok and Mcl-1 were down regulated in all stages of specimen, but Bmf in group II and BBC3 in group III were up-regulated. Our current findings demonstrated the involvements of mRNA expression of Bcl-2 family in multi-step tongue carcinogensis. This highlights the necessity for continued efforts to discover suitable biomakers (Bcl-2 family) for early diagnosis of the disease, and to understand its pathogenesis as a first step in improving methods of treatment. The discovery of these potential biomarkers and molecular targets for cancer diagnostics and therapeutics has the potential to significantly change the clinical approach and outcome of the disease. 전 세계적으로 구강암의 빈도는 점점 증가 추세이며, 특히 한국인의 있어 혀(tongue)는 구강암이 가장 호발하는 장소이다. 구강암은 발암 단계에서부터 과증식 병소(hyperplastic lesion), 이형성(dysplasia) 및 상피내암(carcinoma in situ) 을 거쳐 악성 암종으로 발전하는 다단계 발암과정을 보이며, 분자 생물학적 변이가 구강암을 진행시킴이 널리 알려져 있다. 또한, 구강암은 일반적으로 암세포의 증식 및 고사(apoptosis)의 억제가 중요한 역할을 하고 있다 알려져 있다. 그리고, Bcl-2 family 는 세포 고사에 주요한 역할을 하고 있음이 알려져 있다. 그러나, 이들과 관련한 구강암 발생과정의 변화에 대해서는 널리 연구된 바가 없다. 본 연구는 백서에서 발암 물질인 4-NQO로 구강암을 유도시키고, 구강암 발생 다단계별로 Bcl-2 family의 mRNA 변화를 RT-PCR을 이용해 살펴보았다. Bcl-2 family는 크게 3군, 즉 1) anti-apoptotic, 2) pro-apoptotic, 그리고 3) BH3 only protein으로 분류할 수 있으며, 본 연구에서 anti-apoptotic molecules인 Bcl-w는 모든 군에서 발현이 감소되었으며, Bcl-2는 발현이 증가 되었다. pro-apoptotic molecules에서는 Bad가 제 3군 (편평세포암종)에서 발현이 증가 되었고, 나머지는 감소하였다. BH-3 only protein에서는 Bmf가 제 2군에서, BBC3가 제 3군에서 발현이 증가하였고, 나머지는 모든 군에서 감소하였다. 결론적으로, 4-NQO로 유도된 백서의 발암단계에서, Bcl-2 family의 mRNA 양상은 다양하게 관찰되었으나, Bad 및 BBC3 mRNA가 제 3군에서, Bmf mRNA가 제 2군에서의 발현이 특별함을 알 수 있어, 다단계 발암과정에서의 구강암을 진단하는데 유용하리라 사료된다.

사람 치은모세포에서 다양한 세포외기질 분자에 light-emitting diode 조사 효과

박형근,장윤영,정권,고미경,장미선,국은별,정민아,임원봉,임채광,김옥준,최홍란 대한구강악안면병리학회 2006 대한구강악안면병리학회지 Vol.30 No.3

Expression of Bcl-2 Family in 4-Nitroquinoline 1-Oxide-Induced Tongue Carcinogenesis of the Rat

최재욱,정성수,이금숙,김병국,김재형,국은별,장미선,고미경,정권,최홍란,김옥준,Choi, Jae-Wook,Chung, Sung-Su,Lee, Geum-Sug,Kim, Byung-Gook,Kim, Jae-Hyeong,Kook, Eun-Byul,Jang, Mi-Sun,Ko, Mi-Kyeong,Jung, Kwon,Choi, Hong-Ran,Kim, Ok-Joon Korean Academy of Orofacial Pain and Oral Medicine 2005 Journal of Oral Medicine and Pain Vol.30 No.3

The number of patients with tongue carcinoma is increasing rapidly among young individuals in many parts of the world. Oral carcinoma progresses from hyperplastic lesion through dysplasia to invasive carcinoma and the concept of "field cancerization" with molecular alteration has been suggested for oral cavity carcinogenesis. Significant improvement in treatment and prognosis will depend on more detailed understanding of the multi-step process leading to cancer development. To induce tongue carcinoma in rat by 4-NQO, each drinking water was made to 10 ppm, 25 ppm, 50 ppm and control (only D.W. without 4-NQO). Specimens were classified into 4 groups such as control, I (mild & moderate dysplasia), II (severe dysplasia and carcinoma in situ), III (carcinoma). The mRNA expressions of Bcl-2 family were evaluated by RT-PCR technique. For anti-apoptotic Bcl-2 family, mRNA expression of Bcl-w was down-regulated in all stages of tongue carcinogenesis model. However, mRNA expression of Bcl-2 was up-regulated. For pro-apoptotic Bcl-2 family, all members were down-regulated in all stages of tongue carcinogenesis model except for Bad mRNA in group III. In terms of BH3 only protein, mRNA expressions of Bok and Mcl-1 were down regulated in all stages of specimen, but Bmf in group II and BBC3 in group III were up-regulated. Our current findings demonstrated the involvements of mRNA expression of Bcl-2 family in multi-step tongue carcinogensis. This highlights the necessity for continued efforts to discover suitable biomakers (Bcl-2 family) for early diagnosis of the disease, and to understand its pathogenesis as a first step in improving methods of treatment. The discovery of these potential biomarkers and molecular targets for cancer diagnostics and therapeutics has the potential to significantly change the clinical approach and outcome of the disease.