망막모세포종 환자의 말초혈액에서 RB1 유전자 돌연변이

박준성,추영애,김대광 대한해부학회 2002 Anatomy & Cell Biology Vol.35 No.4

피부 각질화세포에서 자외선 B 조사로 발현이 유도된 유전자의 검출

노진석,이인환,추영애,정수경,최인장,김홍태 계명대학교 의과학연구소 2001 계명의대학술지 Vol.20 No.1

Ultraviolet light is the most important environmental insult to skin. Even single exposure to UVB irradiation can result in inflammation and may also lead to DNA damage and apoptosis in the acute response of the cutaneous tissue. To elucidate the complex alterations of gene expression in human keratinocytes underlying these UVB responses, ordered differential display polymerase chain reaction (ODD-PCR) technology was performed in one human keratinocyte cell line. Several genes were induced and down-regulated during 12 hours after UVB irradiation in a time-dependent manner. Five UVB-induced cDNA clones were isolated, including cDNAs for karyo-pherin alpha 2, human mRNA for TI-227H, ribosomal protein L13a and keratin 19. Differential expression of these genes after UVB irradiation has not been reported. Authors identified a new sequence that is negatively regulated by UVB irradiation. In general our results showed that ODD-PCR is a useful tool in the analysis of quantitative changes of mRNA levels in human keratinocytes after UVB irradiation. The identification of new UVB-modulated genes offters the opportunity to identify unrecognized molecular mechanism in response of human cells to UV irradiation

사람 피부각질세포에서 자외선 B조사로 유도된 핵형 및 세포주기

차호,송인환,추영애,박속경,이인환 대한해부학회 2000 Anatomy & Cell Biology Vol.32 No.2

[지상1]Bcl-2 Overexpression in Acute Lymphoblastic Leukemia Cell Line with t(18;21)

김대광,박형태,추원호,정수경,추영애,김흥식,전동석 대한체질인류학회 2002 대한체질인류학회 학술대회 연제 초록 Vol.45 No.-

망막모세포종 환자의 말초혈액에서 RB1 유전자 돌연변이

박준성(Jun-Sung Park),추영애(Young-Ae Choo),김대광(Dae-Kwang Kim) 대한해부학회 2002 Anatomy & Cell Biology Vol.35 No.4

망막모세포종은 RB1 유전자의 돌연변이에 의해 눈에 생기는 악성 종양으로, 우리나라와 서구에서의 발병율은 서로 비 슷하다. 망막모세포종은 비유전성과 유전성으로 분류할 수 있으며, 유전성인 경우에는 이차 종양으로 뼈근육종이 동반된 다. 유전성 망막모세포종의 약 10~12%는 종양이 한쪽 눈에 생기거나, 먼저 한쪽 눈에 발병한 다음 양쪽 눈으로 발전하는 경우도 있기 때문에, 망막모세포종에서 유전성과 비유전성의 구분과 적절한 치료와 추적을 하기 위해서는 정확한 RB 유 전자의 돌연변이 검사법이 필요하다. PCR-SSCP 방법을 이용하여 엑손 8, 10, 11, 14~20, 22 및 23 부분에 대한 RB1 유전자의 돌연변이를 한쪽 눈에 생기고 가족력이 없는 5명의 망막모세포종 환자의 혈액에서 DNA를 추출하여 돌연변이를 조사 하였다. 엑손 8부분에서 SSCP 띠의 이동 변화가 3예에서 관찰되었으며 염기서열분석 결과 251 코돈에서 CGA → TGA (arginine → 종결코돈)의 점돌연변이가 확인되었다. 우리나라의 보고에는 아직 엑손 8에서 CGA→TGA 돌연변이는 검출되 지 않았으나, 국내의 망막모세포종의 발병율이 서구와 비슷하게 나타나므로 향후 더욱 더 많은 사례를 조사하면 외국의 논문 결과와 같이 엑손 8에서 많은 돌연변이가 나타날 것으로 추정된다. 나아가서 우리나라에서 산발적으로 발생하는 망 막모세포종 뿐만 아니라 유전성 망막모세포종을 지닌 환자와 그 자녀에서 정확한 진단과 치료 방향을 결정하기 위해서는 지속적인 RB1 유전자의 이상에 대한 연구가 필요할 것으로 생각된다. Retinoblastoma, a child tumor of the eye, is caused by two mutational events at the retinoblastoma gene (RB1). Retinoblastoma occurs in both hereditary and nonhereditary forms, and this distinction has important implications for patients and their families. In most patients with isolated unilateral retinoblastoma, tumor development is initiated by somatic inactivation of both alleles of the RB1 gene. Some of patients with hereditary retinoblastoma initially present with unilateral disease, and up to 10% to 12% of these patients only express unilateral disease. Germline mutation in RB1 gene confer hereditary predisposition to retinoblastoma. This study was designed to identify germline mutations in RB1 gene in Korean retinoblastoma patients. Samples of peripheral blood were obtained from 5 patients with isolated unilateral tumors. To detect genetic alteration in RB1 gene, exon 8, 10, 11, 14~20, 22 and 23 were investigated by PCR-SSCP. Bandshifts on SSCP were found in three out of 5 patients at exon 8. There were same point mutations from CGA (arginine) to TGA (stop codon) at codon 251 in exon 8 of RB1 gene. This point mutation has not been found in Korean patient with retinoblastma. But it is common mutation on the Western reports and Korea’s annual incidence of this tumor is similar in proportion to that of the West. Therefore, if a lot of patients are investigated to elucidate RB1 mutation this point mutation will be found. Identification of the germline mutation in RB1 gene would help to improve the presymptomatic diagnosis and clinical management to retinoblastoma patients.

자외선 B 조사에 의한 사람 피부 각질화세포의 분자생물학적 반응

황성택,손수상,이인환,추영애,김홍태 대한체질인류학회 2001 대한체질인류학회 학술대회 연제 초록 Vol.44 No.-

여성 생식호르몬 영향에 의한 자궁내막과 자궁내막암에서 텔로메레이즈 ( TELOMERASE ) 의 활성

서성일,이태성,차순도,조치흠,추영애 대한부인종양 콜포스코피학회 1999 Journal of Gynecologic Oncology Vol.10 No.1

During the reproductive period, human endometrium undergoes a pattern of cyclic change. Human endometrium undergoes a complex pattern of proliferation, secretory activity, and menstruation over an approximately 28 days period. Proliferative activity is highest during late proliferative phase under influence of estrogen, and minimal activity in the late secretory and menstrual phase. To study a possible change of telomerase activity during menstrual cycle, telomerase activities in normal and hormone treated endometrium were tested using telomerase repeat amplification protocol(TRAP) assay. Telomerase activities were detected in 9 of 10 proliferative endometrium(90%), and maximal activity was shown in late proliferative phase. Only 3 of 10 secretory endometrium(30%) revealed weak activity. However telomerase activity was not detected in menstrual phase endometrium(N 2) and senile endometrium(N=3). Four of tamoxifen treated endometrium(N 4) and 1 of provera treated endometrium(N 3) Levels of telomerase activity of treated endometrium(N 4) and late proliferative endometrium(N 6) were as high as them of detected in endometrial cancer and hyperplasia. Above findings reveal that telomerase activity of endometrium is changed according to menstrual cycle, And the level of telomerase activity is related to proliferative activity of endometrium that is dependent on the status of female sex steroid hormone and tamoxifen treatment. $quot;

자궁경부 상피세포암의 암화과정에 따른 인유두종 바이러스 감염과 Telomere 길이 및 Telomerase의 활성

서민호,이태성,박종하,차순도,조치흠,추영애,백원기,서성일 대한부인종양 콜포스코피학회 1997 Journal of Gynecologic Oncology Vol.8 No.1

E6 and E7 proteins produced by oncogenic HPV bind to the protein products of cellular tumor suppressor genes p53 and Rb, respectively. This mechanism has been suggested to contribute to the oncogenesis of HPV-infected carcinoma. The cells which are blocked the function of p53 and pRb protein continue to divide by bypassing M1 stage known as antiproliferative mechanism but telomeres, the genetic elements at the ends of chromosomes, continue to shorten until the telomeres are so short that further replication is prevented(M2 stage). But telomeres can be maintained if telomerase is derepressed, giving rise to a immortal cell. The present study has been investigated the presence of HPV, telomere length and telomerase activation in cervical carcinomas. HPV DNA were detected by polymerase chain reaction in 17 of 19 precancerous lesions and cervical carcinoma specimens; HPV16 was detected in 12 cases, HPV18 in one case, HPV33 in two cases, and HPV58 in two cases. Overall, the prevalence of HPV was 89.5%. To study the difference of telomere length in cervical carcinomas and each normal counterpart, DNAs were digested with Hinfill and Rsal to liberate the terminal restriction fragments(TRF). TRFs were resolved on agarose gels and detected by hybridization to the telomeric probe. This result indicated that there were no significant difference of TRF length in samples tested expept two cases. TRF length of one carcinoma specimen was found to be significantly increased as compared with normal counterpart, but the other was found to be significantly decreased. Telomerase activity was detected in 4 of dysplasia specimens(5 cases), all of carcinoma in situ(CIS), and 6 of 8 invasive carcinoma. Overall, telomerase activity was detected in 84%. The degree of telomerase activity was high in 2 of dysplasia, 3 of CIS, and 3 of invasive carcinoma. And then there was no apparent association between HPV types and levels of telomerase activity. However, telomerase activity was depressed in invasive carcinoma as compared to dysplasia and CIS. These results suggest that HPV may be a possible causative agent in cervical carcinoma. In addition, telomerase activation may be necessary for the immortalization of cells and the progression of malignancy in cervical carcinoma.

자궁근종에서 도파민수용체와 전달체의 발현 증가와 도파민전달체 유전자의 메칠화 감소

김주현 ( Ju Hyun Kim ),김민지 ( Min Ji Kim ),추영애 ( Young Ahe Choo ),최윤석 ( Yoon Suk Choi ),이태성 ( Tae Sung Lee ),김홍태 ( Hong Tae Kim ) 대한산부인과학회 2005 Obstetrics & Gynecology Science Vol.48 No.3

목적: 도파민은 민무늬근육세포의 증식을 조절함에 있어 중요한 역할을 한다. 이 연구에서는 자궁근종과 정상자궁근육조직에서 도파민 D1과 D2 수용체와 도파민전달체 (DAT)의 발현정도와 DAT 발현 조절에 메칠화의 작용을 알아보고자 하였다. 연구 방법: 자궁근종으로 자궁절제술을 받은 20명의 환자로부터 정상자궁근육과 자궁근종조직을 얻었다. 이들 조직에서 도파민 D1과 D2 수용체와 DAT의 발현정도를 RT-PCR과 면역조직화학법으로 조사하였다. DAT 유 Objective: Dopamine plays a key role in the proliferation regulation of the smooth muscle cells. The purpose of this study was to observe the degree of expression of dopamine D1 and D2 receptors and dopamine transporter (DAT) and to evaluate the influence

사람 피부각질세포에서 자외선 B 조사로 유도된 핵형 및 세포주기 변화

차 호(Ho Cha),송인환(In Hwan Song),추영애(Young Ae Choo),박숙경(Sook Kyung Park),이인환(In Hwan Lee) 대한해부학회 2000 Anatomy & Cell Biology Vol.33 No.1

자외선 B 의 조사로 인하여 유전자가 표적이 되어 세포 주기가 변화하고 그후 증식하는 과정을 조사하기 위하여 자외 선 B 200J/m2를 조사한 후 흘림세포계측기 (flowcytometer)에서 CellFit 프로그램으로 세포주기의 각 기를 측정하였다. 주된 세포 주기의 변화는 S기가 길어진 것이었으며 핵형 분석에서는 염색체의 수가 near diploid (43-47)에서 hypotetraploid(80+)로 되었다. 자외선 B를 조사한 후 near diploid (43-47)군과 hypotetraploid (80+)군으로 나누어 세포주기를 측정하였더니 near diploid군에서는 S기의 연장이 특징적이었으며 hypotetraploid군에서는 G0-G1기의 길어짐을 보였으며 세포 배양에서 형질 전형을 보였다. 핵형 분석에서 나타나는 새로운 구조적 변형은 del (5q21), 8p+ 및 t (5 : 8) (q21 : pter)이었다. 이중 del (5q21)이 모든 형질 전형된 세포에서 나타났다. 이상의 결과로 미루어 자외선 B의 조사로 변화하는 세포 주기는 M기를 한번 거치지 않는 [G1-S-(G2-G1)-S]의 과정으로 여겨지며 del (5q21) 자리는 암 억제유전자인 APC 유전자의 위치여서 이 유전자의 소실이 세포배양에서 배양 접시에 붙지 않고 자라는 형질전형의 주된 원인이 되는 것으로 생각된다. 핵의 크기 변화는 세포 주기의 변화로 인한 염색체 수의 변화에서 기인되며 세포의 수적 증가가 늦어지는 것으로 생각된다. To investigate the change of cell cycle by genotoxic stress and rebound proliferation in human keratinocytes, the proportions of cell cycle phases were estimated with challenge of UVB irradiation (200 J/m2). With UVB irradiation cell cycle was estimated by Cell Fit program in Flowcytometer, and main change of the cell cycle was S-phase prolongation. In karyotyping, near diploid number of chromosomes changed to hypoteraploid number. Cell cycle phase was estimated in two groups of cells, near diploid and hypoteraploid. In near dipoid cells, S-phase prolongation was specific phenomenon, while specific G0-G1 phase prolongation was shown in hypoteraploid cells which made transformed foci in culture. The new structural anomalies were del (5q21), 8p+, and t (5 : 8)(q21 : pter). Among them, del (5q21) was found in all transformed hypotetraploid cells. These data suggest that progress of cell cycle could be [G1-S-(G2-G1)-S] by UVB irradiation and deletion of 5q21 has a key role for anchorage independent growth, which is deletion of tumor suppressor gene APC locus. That is one of important mechanisms in keratinocyte transformation by UVB irradiation. With the changes of chromosome number and cell cycle, sizes of nuclei got to bigger by two times and growth rate was delayed.