자궁경부암 환자에 있어 telomerase 활성도의 임상측명

조삼현(SH Cho),유중배(JB Yoo),김승룡(SR Kim),김기성(KS Kim),라명재(MJ Ra),김경태(KT Kim),문형(H Moon),황윤영(YY Hwang) 대한산부인과학회 1997 Obstetrics & Gynecology Science Vol.40 No.4

A current hypothesis gaining prominence proposes that activation of the telomerase is necessarey for cells to become immortal, or be capable of proliferating indefinitely. The theory suggersts that that almost all cancer cells must attain immortality for progression to malingnant state and, hence, require activation of telomerase. To assess the role of telomerase in the development of mailignant transformation of the uterine cervical carcinoma, telomerase activeiy was measured by using a recently developed sensitive RPR-based telomerase assay(telomeric repeat amplification protocol; TRAP)in benign uterine condition and invasive uterine cervical carcinoma tissues revealed to be telomerase positive (14/16, 88%) which represents a characteristic 6 bp ladder pattern, while none of the 4 cervical tissues with the nonspecific pathologic findings or just chronic cervictis which were obtained by simple bysterenctomy under the diagnoses of benign myoma or uterine prolapse showed any telomerase actiovity. These findings suggest that this enzyme activity may play a key role in the establishment and progression of the uterine cervical carcinoma. This activity of telomerase may influence from the initial stage of tumorigenesis of uterine cervical carcinoma because poitive findings were noticed from the early stage (stage (i 1a1) to the advanced stage. The results of neoadjuvant chemotherpy which were revealed in final histopathologic specimens following radical hysterectomuy, pelvic and raraaortic lymph nodes dissection (suggestion by Rosen et al,) were one grade IV wiht telomerase poistive, 2 grade III with all positivie, 5grade II with 3 positive 2 negative. The two cases of pelvic lymph node metastasis belong to the group of grade II and their activeities of telomerase were 1 positive and 1negative. The situation of apoptosis induced by antineoplastic chemotherapy has not been related to that of telomerase activeity. Furthemore, the late recurrences has been noticed over 5years follow-up following neoadjuvant chemotherapy and radical surgery for the management of the high risk group of patients (not shown in this study) This measn that the nowday`s regimen for induction tumoricidals may have been operating as dual effects (partial restoring and redamaging) to the cell cycle chekpoint. Therefore if telomerase inhibitor will be applied combined whit the induction chemotherapy, unexpected benefits mignt be obtained for the patient with the advanced uterine cervical carcinoma.

자궁경부암에서의 Telomerase 활성도

이경아(Kyung Ah Lee),장태기(Tae Kee Jang),장영진(Young Jin Jang),이영기(Young Gi Lee),이두진(Doo Jin Lee),이승호(Sung Ho Lee) 대한산부인과학회 2000 Obstetrics & Gynecology Science Vol.43 No.2

Objective: Telomerase is a ribonucleoprotein that synthesizes TTAGGG repeats onto chromosome ends. The expression of telomerase is thought to be required for cellular immortality and carcinogenesis. This study was conducted to examine the telomerase activation occurs in cervical carcinogenesis. Methods: The standard telomeric repeat amplification protocol(TRAP) was used to examine telomerase activity in tissues of 10 normal cervix, 10 carcinoma in situ, and 21 invasive cervical carcinoma. Results: Telomerase activity was detected in tissues of 16/21(76.2%) invasive carcinoma, in 5/10(50.0%) carcinoma in situ, and in 3/10(30.0%) normal cervix. But the degree of telomerase activity in normal cervix was weak. There was significant difference in 3 groups(p<0.05). The results of neoadjuvant chemotherapy in 10 invasive cervical carcinoma were as follows. In 8 cases of which tumor size decreased more than 50%, 5 were positive for telomerase. In 2 cases that didn't respond to chemotherapy by tumor size, 1 was positive for telomerase. There was no significant difference between 2 groups. All of the 5 cases that had pelvic lymph node metastasis revealed positive telomerase activity, and the 11 cases of 16 cases that didn't have pelvic lymph node metastasis were positive for telomerase, but there was no significant difference in 2 groups. The positivity of telomerase activity in clinical stage of invasive cervical carcinoma was 73.3% in stage I(11/15), 75.0% in stage II(3/4), 100% in stage III(1/1), and 100% in stage IV(1/1), but there was no significant difference in each stages. Conclusion: Telomerase seems to be uniquely associated with malignant transformation of cervix and can be used as a tumor marker. Additional studies are needed to better clarify the biological significance of telomerase expression in cervical tumorigenesis.

HL-60 백혈령 세포주에서 분화유도물질로 전처치 후의 telomerase 활성

김인호,김숙자,정희정,박성규,이규택,원종호,서원석,백승호,홍대식 大韓血液學會 1999 大韓血液學會誌 Vol.34 No.1

난소암에서의 텔로메레이즈 ( Telomerase ) 활성도의 변화 탐색

송진화(JH Song),조치흠(CH Cho),차순도(SD Cha),이태성(TS Lee) 대한산부인과학회 1998 Obstetrics & Gynecology Science Vol.41 No.4

Telomerase is a ribonucleoprotein that can add hexameric TTAGGG sequences to the end of chromosomes and thus stabilize telomeres. Telomeres are shortened by progression of cell division, a process known as cellular senescence. If telomerase activity is not activated the chromosomes are shortened to a critical length and the division of cell is stopped. In cancer cells telomere shortening is stopped by the prescence of the enzyme telomerase and the cells divisions is contined indefinitely. Most normal somatic tissues don`t express telomerase activity, but both fetal and adult human ovaries and testis express telomerase. To investigate the relationship between telomerase activity and acquisition of malignancy in ovaries, the activity of telomerase was assayed in normal, benign, borderline and malignant ovarian tissues. With TRAP assay, the telomerase activity was examined in 15 normal ovarian tissues, 1 benign mucinous cystadenoma, 1 borderline carcinoma, and 10 ovarian malignancies. Telomerase activities were detected weakly in 9 of 15 cases of normal ovarian tissues and were not related to the activity of reproduction. The strong activity of telomerase were found in borderline carcinoma and 5 of 6 cases of epithelial ovarian carcinoma and 1 rest case revealed low activity. Telomerase activities in dysgerminoma were undetectable in one and weak in rest one, but 1 case of immature teratoma revealed strong activity. The telomerase is reactivated or upregulated in epithelial carcinoma of the ovary, as it is detectable in borderline carcinoma, but is undetectable or weakly detectable in benign ovarian tumor and normal ovarian tissues. Telomerase activity is closely related to the epithelial ovarian carcinoma, and it may be useful as a marker for diagnosis and as a target for therapeutic intervention. The dysgerminoma revealed no or weak activity of telomerase compared to another germ cell tumors and epithelial carcinomas, so further study will be needed to examine the role of telomerase activity in dysgerminoma.

기초 : 뇌종양에서 진단적 지표로서의 Telomerase의 활성과 그 조절 기전에 관한 연구

조창원 ( Chang Weon Cho ),최창화 ( Chang Hwa Choi ),차승헌 ( Seung Heon Cha ),김철민 ( Cheol Min Kim ) 대한뇌종양학회·대한신경종양학회·대한소아뇌종양학회 2005 대한뇌종양학회지 Vol.4 No.2

Objective￡ºTelomerase is the ribonucleoprotein complex which elongates telomeric repeats(TTAGGG)n and appears to play an important role in cellular immortalization. The almost exclusive expression of telomerase in tumor cells, and not in most normal cells, offers an exciting opportunity for therapy by inhibiting its function. In order to know the possibility of telomerase activity as a diagnostic marker for prediction of prognosis in brain tumors, telomerase activity was investigated in and cell lines. Patients and Methods￡ºThe surgical specimens from 34 patients with of brain tumors were analyzed by TRAP assay for access telomerase activity. And the mechanism of regulation on telomerase activity in brain tumor was investigated by RT-PCR analysis of catalytic subunit of telomerase gene, hTERT, and c-myc oncogene in five cell lines. To investigate more factors involved in carcinogenosis in malignant brain tumors, gene expression analysis with Atlas human cDNA expression array was conducted in two glioblastoma cell lines, A172 and U373MG. Results￡ºMalignant brain tumors have high activity of telomerase. This means telomerase activity can be used as a marker for malignant brain tumor. The catalytic subunit of telomerase, hTERT, were coamplified with c-myc oncogene in brain tumors with high activity of telomerase. This finding suggest the possibility of c-myc as a regulatory factor on telomerase function. Atlas human cDNA expression array profiles of two cell lines showed common expression of 27 genes which were known as genes involve during carcinogenesis. Among 27 genes, there were four genes of cell cycle or growth regulators, one intermediate filament marker, three genes for apoptosis, two genes for DNA damage repair, three receptor genes, four genes for cell adhesion and motility, one gene for angiogenesis, one genes of invasion regulators, one gene belong to Rho family of GTPases, and seven genes of growth factors and cytokines. Conclusion￡ºThis study suggest that c-myc might be a possibile upstream regulator of telomerase activity in brain tumors and that 27 genes might involve the carcinogenosis of malinant brain tumor especially in glioblastoma.

정상 및 각종 혈액종양 질환의 조혈세포에서의 Telomerase 활성도

안명주,노연희,임그리워,박일규,이웅수 大韓血液學會 1996 大韓血液學會誌 Vol.31 No.3

방광암 환자에서 Telomerase PCR-ELISA 검사법

이선주,이우인,장성구 大韓泌尿器科學會 1999 Korean Journal of Urology Vol.40 No.3

기초 :뇌종양 조직으로부터 Telomerase의 순수분리와 활성 억제에 관한 연구

황순구 ( Soon Gu Hwang ),이영우 ( Young Woo Lee ) 대한뇌종양학회·대한신경종양학회·대한소아뇌종양학회 2009 대한뇌종양학회지 Vol.8 No.1

Objective:Telomerase activity was examined in human brain tumor and compared with their histological diagnosis. Methods:The 70 brain tumor tissues were surgically obtained from 68 cases at Department of Neurosurgery, Pusan National University Hospital between 1996 and 1999. Results:The result of telomerase assay of various brain tumor tissues, 6 of 8 glioblastoma multiforme, 7 of 8 anaplastic astrocytoma, 1 of 1 anaplastic oligodendroglioma and 5 of 6 metastatic brain tumor, demonstrated telomerase activity. But epidermoid tumor, cystic teratoma, pituitary adenoma, ependymoma and meningioma did not exhibit high activity. Immunohistochemistry study using anti-CD68 primary antibodies was performed and the proliferative activity of tumor cells were evaluated. By immunohistochemistric study and then, the mean CD68 proliferative cell indexes(PCI) were the followings: glioblastoma(38.4%), anaplastic astrocytoma(18.0%) and anaplastic oligodendroglioma(16.70%). The results recognized that there was no correlation between telomerase activity and CD68 PCI. But the malignant brain tumors had higher PCI values than those of benign tumors. The telomerase from brain tumor cells was purified to homogeneity through one step affinity column chromatography using Ultralink- immorbilized Neutravidin plus beads. The resulting affinity purified telomerase was active in PCR-amplified telomerase assays. The purified telomerase complex was a molecular weight of 374 KD and had triple components. The effect of antisense oligonucleotides complementary to the RNA component of telomerase on telomerase activity in purified enzyme had been studied. Pronounced inhibition of telomerase activity was observed at the oligonucleotide concentration in the reaction mixture of about 0.4 μM. Conclusion:It might be considered that the antisense oligonucleotide complementary to the human telomerase RNA component in the region of the template synthesis of telomeric repeats is the efficient inhibitor of telomerase activity.

성숙 백서 음경에서 남성호르몬 변화에 따른 Telomerase활성도의 영향

최영득,박진아,마상열,이무상,최형기,최영득,박진아,마상열,이무상 大韓泌尿器科學會 1999 Korean Journal of Urology Vol.40 No.7

아데노 바이러스를 이용한 Telomerase-Antisense 발현이 조혈모세포 체외 배양에 미치는 영향

김석진,김병수,오상철,서재홍,최철원,신상원,김열홍,김준석,김영태,송준석,최은정 대한조혈모세포이식학회 2003 대한조혈모세포이식학회지 Vol.8 No.1

연구배경: 조혈모세포는 자가갱신능 및 분화능을 지니고 있지만, 조혈모세포의 체외배양을 통한 자가갱신을 유도하는 기술 및 정상 혈액세포로 분화, 증식시키는 기술이 개발되어 있지 않고 있다. 그런데, 줄기세포의 자가갱신에 telomerase가 매우 중요한 역할을 수행하고 있음이 알려져 있고 이에 저자 등은 조혈모세포를 telomerase antisense로 처리 후 체외 배양을 하였을 때 어떤 결과가 초래되는지를 알아보고자 본 연구를 시행하였다. 방법: 총 8명의 정상인 골수 공여자에서 채집한 골수혈로부터 단핵구를 분리 후 1×10^(5)/ml의 농도로 LTC-IC 배양을 한 대조군과 단핵구(1×10^(5)/ml)를 telomerase antisense (Adenovirus, 10⁴ PFU/ml, 3 ml) 처리 후 LTC-IC 배양을 한 실험군으로 나누어 양군간의 배양 2주 후 세포 수, 생존율, 그리고 세포모양을 비교, 분석하였다. 결과: LTC-IC 배양 2주에 계산한 세포 수의 경우, 대조군은 1.08±0.19×10⁴/ml인 데 반하여 실험군은 6.75±2.60×10⁴/ml로 통계적으로 유의하게 실험군에서 세포 수가 많았다(P=0.001). LTC-IC 배양 2주에 관찰한 세포 모양의 경우 대조군은 모든 세포들이 일정한 크기로 구형을 유지하고 있었으나 실험군에서는 크기가 서로 상이하고 분화된 세포들이 상당 부분에서 관찰되었다. Tryphan dye로 염색 후 100개의 세포를 관찰하여 염색되지 않은 세포를 생존 세포로 간주하여 그 백분율을 계산하여 보았다. 그 결과 대조군은 83.8±4.8%이고 실험군은 81.1±2.1%로서 통계적으로 유의한 차이는 없었다(P=0.288). 결론: 조혈 모세포를 Telomerase Antisense로 처리 시 분화가 촉진될 가능성을 본 실험을 통하여 확인할 수 있었다. 그러나 본 실험에서는 Telomerase Antisense로 처리 후 LTC-IC 배양으로 증식된 세포들의 성상 하나, 하나를 확인하지 못하였다는 한계가 있었다. Background: Hematopoieic stem cells are characterized by their ability of self-renewal and differentiation into mature blood cells. Telomerase, which is known as a target of anti-cancer therapy, is also regarded as having possibility of inducing self-renewal of hematopoietic stem cells, and inhibition of telomerase activity might be related with differentiation of hematopoietc stem cells. Thus, we evaluated how telomerase antisense oligonucleotide could affect hematopoietic stem cells in vitro culture. Methods: We used adenovirus vector containing recombinant antisense telomerase template RNA. Mononuclear cells were obtained from 8 bone marrow donors and divided into two groups. Using the long-term culture system, the experiment group was cultured with treatment of telomerase antisense oligonucleotides, while control group was not. We evaluated the cell number and the degree of differentiation at 2 weeks after LTC-IC culture. Results: The mean cell number of LTC-IC culture was more higher in the telomerase antisense treatment group than that of control group (6.75×10⁴±2.60/mL vs 1.08×10⁴±0.19/mL). On microscopic examination, the cells of telomerase antisense treatment group showed more differentiation pattern than the group of the control group. Conclusion: We found that adenovirus-mediated antisense expression of telomerase RNA might influence the differentiation of hematopoietic stem cells. However, further study should be warranted to evaluate its consequences and mechanisms.