급성 골수성 백혈병에서 Fas 및 Bcl-2 항원의 발현과 그 임상적 의의

이석,민유홍,정소영,김성철,유내춘,이정윤,권오현,한지숙,고윤웅 大韓血液學會 1996 大韓血液學會誌 Vol.31 No.3

HS-1200 Overcomes the Resistance Conferred by Bcl-2 in Human Leukemic U937 Cells

Jun-Young Park,Jeong-Bon Moon,In-Ryoung Kim,Gyoo-Cheon Kim,Bong-Soo Park,Hyun-Ho Kwak KOREAN ACADAMY OF ORAL BIOLOGY 2012 International Journal of Oral Biology Vol.37 No.3

Bcl-2 protects tumor cells from the apoptotic effects of various anti-neoplastic agents. Increased expression of Bcl-2 has been associated with a poor response to chemotherapy in various malignancies, including leukemia. Hence, bypassing the resistance conferred by anti-apoptotic factors such as Bcl-2 represents an attractive therapeutic strategy against cancer cells, including leukemic cells. This study was undertaken to examine whether the anticancer drug, cisplatin and the synthetic chenodeoxycholic acid (CDCA) derivative, HS-1200 show anti-tumor activity in U937 and U937/Bcl-2 cells. Viability assays revealed that HS-1200 overcomes the resistance conferred by Bcl-2 in human leukemic U937 cells. Various apoptosis assessment assays further demonstrated that HS-1200 overcomes the resistance conferred by Bcl-2 in human leukemic U937 cells by inducing apoptosis. In addition HS-1200, but not cisplatin, overcomes the anti-apoptotic effects of Bcl-2 in Bcl-2 over-expressing human leukemic cells (U937/Bcl-2 cells). Notably, we observed that the HS-1200-induced formation of mature promyelocytic leukemia (PML) nuclear bodies (NBs) correlates with a suppression of the anti-apoptotic effects of Bcl-2 in human leukemic cells over-expressing this protein (U937/Bcl-2 cells). Furthermore, HS-1200 was found to induce the association between PML and SUMO-1, Daxx, Sp100, p53 or CBP in the aggregated PML-NBs of U937/Bcl-2 cells. Thus, PML protein and the formation of mature PML-NBs could be considered as therapeutic targets that may help to bypass the resistance to apoptosis conferred by Bcl-2. Elucidating the exact mechanism by which PML regulates Bcl-2 will require further work.

대장 선종-선암에서 bcl-2종양 단백 발현에 대한 연구

문철,김정욱,조준형,허철행,김형준,도재혁,장세경 大韓消化器病學會 1998 大韓消化器病學會誌 Vol.31 No.2

자궁경부 상피내종양 및 자궁경부암 조직에서 bcl-2 및 c-myc 암유전자 발현과 세포증식 및 apoptosis와의 상관관계에 관한 연구

김병기(Byoung Gie Kim),이효표(Hyo Pyo Lee) 대한산부인과학회 1999 Obstetrics & Gynecology Science Vol.42 No.8

bcl-2 암유전자는 여러 가지 자극에 의한 apoptosis를 차단함으로써 유전자 이상을 가진 세포가 계속 생존하면서 유전자 변이가 누적되는 결과를 초래한다고 알려져 있다. 한편 c-myc 암유전자는 세포증식과 apoptosis를 유도하는 이중적 기능을 가지고 있으며 생존 신호가 결여될 경우에는 오히려 세포의 apoptosis를 유발한다고 알려져 있다. 그러나 c-myc과 bcl-2가 동시에 발현되면 bcl-2는 c-myc의 세포증식 작용은 영향을 주지 않고 apoptosis만을 선택적으로 차단함으로써 유전자 변이 세포의 생존 뿐만 아니라 증식을 촉진하는 것으로 관찰되었다. 자궁경부암에서 c-myc과 bcl-2 발현에 관한 개별적 보고는 있었으나 이들 두 유전자의 동시발현 및 이들 유전자들이 실제 암조직상에서 세포증식 및 apoptosis에 어떠한 영향을 미치는가에 관한 연구는 보고된 것이 없다. 따라서 본 연구에서는 자궁경부암 발생 과정중에서 bcl-2 및 c-myc 발현과 세포증식, apoptosis와의 상관관계를 알아보고자 하였다. 본 연구에서는 10개의 정상 자궁경부조직, 30개의 자궁경부 상피내종양 조직, 20개의 자궁경부암조직에서 bcl-2와 c-myc에 대한 면역조직화학 검사를 시행하였으며 세포증식과 apoptosis는 각각 Ki-67 면역조직화학적 방법과 TUNEL 방법으로 확인하였다. 또한 환자의 임상병리학적 인자들과의 상관관계도 알아보았다. 정상 자궁경부, 자궁경부 상피내종양, 자궁경부암조직 중 자궁경부암조직에서만 bcl-2와 c-myc 단백이 각각 35%와 50%에서 관찰되었으며, 또한 bcl-2와 c-myc의 동시발현이 25%에서 관찰되었다. 세포증식 지수(상피세포 100개중 Ki-67양성 세포수)는 정상 자궁경부, 상피내종양, 자궁경부암으로 진행되면서 10.2, 24.1, 59.7, 71.2로 유의하게 증가하는 양상을 보였으며(p<0.01), apoptosis 지수(상피세포 100개중 apoptosis 세포수)도 0, 0.33, 1.85, 3.89로 점차 증가하는 양상을 보였다(p<0.01). 또한 세포증식 지수와 apoptosis 지수와는 높은 상관관계(r=0.7451, p=0.0002)를 나타내었다. 그러나 자궁경부암 조직중 bcl-2 발현군과 비발현군간에 apoptosis지수에는 차이가 없었으며(p=0.4765), c-myc 발현군과 비발현군간에도 세포증식 지수에는 차이가 없었다(p=0.6891). 또한 bcl-2와 c-myc의 동시 발현군과 나머지 군간에도 증식지수와 apoptosis 지수에 차이가 없었다(각각 p=0.6311 및 p=0.7600). 한편 bcl-2와 c-myc의 동시 발현과 잘 알려져 있는 자궁경부암의 임상병리학적 예후인자들(종양 크기, FIGO 임상병기, 림프절 전이등)과는 유의한 상관관계가 없었다. 이상과 같은 결과에서 자궁경부암발생 과정에서 세포증식과 apoptosis는 병변의 등급과 비례하여 증가하고 apoptosis는 세포증식과 관련된 변화로 사료되었다. 한편 bcl-2와 c-myc과 발현은 자궁경부암에서만 관찰되는 유전자 변이로서 자궁경부 상피내종양의 발생과 진행과정에는 영향을 미치지 않으며, 또한 자궁경부암 조직에서도 암조직 전체의 세포증식 및 apoptosis와는 관련이 없을 것으로 사료되었다. bcl-2 prevents cell death from a wide variety of stimuli and provides survival of cells with accumulated genetic alterations and c-myc can promote both cell proliferation and cell death through the transcriptional regulation of target genes. Although several studies have been reported on the expression of bcl-2 or c-myc separately, little has been known about the role of coexpression of bcl-2 and c-myc to cell proliferation and apoptosis, as well as the frequency of these coexpression in cervical cancer specimens. In this study, we have examined the expression of bcl-2 and c-myc in cervical cancer specimens and cervical intraepithelial neoplasia(CIN) to determine the role of coexpression of bcl-2 and c-myc during progression into cervical cancer. Proteins and transcripts of bcl-2 and c-myc were evaluated by immunohistochemistry in 60 clinical specimens(20 cervical cancer, 30 CIN, and 10 normal cervix). In addtion, we evaluated kinetic indices of cell proliferation and apoptosis simultaneously. The cell proliferation index was determined by detection of the Ki- 67 in immunohistochemistry. Apoptotic index was determined by the detection of apoptotic cells with TUNEL staining. Medical records including pathologic reports were reviewed. Overexpression of bcl-2 and c-myc was identified in 7(35%) and 10(50%) of 20 cervical cancer specimens respectively, but none in normal cervix and CIN samples. In addition, coexpression of bcl-2 and c-myc was found in 5(25%) of 20 cervical cancer specimens. The cell proliferation index increased with progression from normal to CIN and invasive cancer(normal cervix, 10.2; CIN 1, 24.1; CIN 2/3 59.7; cervical cancer, 71.2; p <0.01). The apoptotic index also increased with grade of lesions(normal cervix, 0; CIN 1, 0.33; CIN 2/3, 1.85; cervical cancer, 3.89; p <0.01) and showed a significant correlation with proliferation index(r=0.7451, p=0.0002). However, there was no significant difference in apoptotic index between bcl-2 positive and bcl-2 negative group in cervical cancer(p=0.4765). In addition, there was also no significant difference in cell proliferation between c-myc positive and c-myc negative group(p=0.6891). Furthermore, there was no significant difference in cell proliferation and apoptosis between bcl-2 and c-myc positive group and others in cervical cancer(p=0.6311 and p=0.7600 respectively). The well-known clinicopathologic parameters, including tumor diameter, FIGO clinical stage, lymph node metastasis, did not correlate with simultanuos positive immunoreactivity for bcl-2 and c-myc proteins in cervical cancer. In conclusion, the cell proliferation and apoptosis increase with increasing lesion grade of cervical neoplasia and apoptosis correlates with cell proliferation. In addition, overexpression of bcl-2 and/or c-myc may be genetic alteration found only in cervical cancer and may not play a role in the development and progression of CIN. However, neither bcl-2 nor c-myc immunoreactivity correlated with the proliferation index or apoptotic index. These results suggest that other factors may also play a role in controlling the cell proliferation and apoptosis of cervical cancer.

기저세포암에서 bcl - 2 와 MIB - 1 단백 발현 양상

이봉길 ( Bong Kil Lee ),이원우 ( Won Woo Lee ),이선경 ( Sung Kyung Lee ) 대한피부과학회 1998 大韓皮膚科學會誌 Vol.36 No.1

Background: bcl-2 is a newly characterized proto-oncogen that has been shown to suppress programmed cell death(apoptosis), which is involved in tumorigenesis, and its expression has been demonstrated within tumor cells in a variety of neoplasms. Objective: We examined whether there are differences in bcl-2 expression in basal cell carcinoma and several epidermal keratinocytic tumors. In addition, we evaluated bcl-2 expression according to histological types of basal cell carcinoma as well as the interaction of expression of bcl-2 and MIB-1 protein in BCC subtypes. Methods : Routine paraffin sections of formalin-fixed sixty tissues(20 BCC, 10 Bowen disease, 10 keratoacanthoma, and 20 SCC) were labelled with anti-bcl-2 monoclonal antibody and MIB-1 using a labelled streptavidin-biotin complex. Results : 1. Twenty cases of basal cell carcinoma were examined and all expressed cytoplasmic bcl-2. Three out of 20 cases of squamous cell carcinoma were focally positive. None of the 10 Bowen disease cases and the 10 keratoacanthoma cases expressed bcl-2. 2. In BCC, bcl-2 immunoreactivity was great in the superficical subtypes which had an indolent growth variant, moderate in the circumscribed types, and weak in the infiltrative types which had aggressive growth variants. 3. Evaluation of the distribution of bcl-2 immunoreactive categories and MIB-1 grades revealed a negative correlation tendency, but no statistical significance. Conclusion : Our results suggest the presence of a protection mechanism from apoptosis mediated by bcl-2 protein involved in the neoplastic growth mechanism of BCC. In addition, the observed findings in the expression pattern of bcl-2 and MIB-1 in the BCC subtypes may be due to interaction between bcl-2 and other apoptotic-related oncogens. (Korean J Dermatol 1998;36(1) 71-77)

Expression of Bcl-2 and Bax mRNAs During Follicular Development and Atresia in the Rat Ovary

고필옥(Phil Ok Koh),강상수(Sang Soo Kang),최완성(Wan Sung Choi),곽수동(Soo Dong Kwak),조경제(Gyeong Jae Cho) 대한해부학회 1999 Anatomy & Cell Biology Vol.32 No.1

포유류의 난소는 생식주기에 따라 난포의 성장 (follicular development)과 퇴화(follicular atresia)가 주기적으로 일어나며 이중 난포의 퇴화는 programed cell death인 apoptosis에 의해 일어난다. Apoptosis는 bcl-2 gene family에 의해 조절되며 이들 유전자 중 bcl-2는 apoptosis를 억제하는 유전자로, bax는 apoptosis를 유도하는 유전자로 알려져 있다. 본 연구에서는 미성숙한 흰쥐에 PMSG를 투여하여 난포의 성장과 퇴화를 유도한 후 이들 난포에서 bcl-2와 bax mRNA의 발현을 in situ hybridization으로 조사하였다. PMSG를 투여한 후 1일과 2일군에서는 대부분의 난포가 난포강을 형성하였고 과립세포가 균일하게 난자를 둘러싸고 있 었으며 투여후 3일군에서는 성장하는 난포와 난포의 형태가 일그러지고 과립세포층이 얇아지는 퇴화하는 난포들이 관찰되었다. 4일, 5일군에서는 난포의 과립세포층이 얇아지고 응축된 핵이 나타난 퇴화하는 난포들이 관찰되었는데 이들 퇴화하는 난포는 in situ DNA end labeling에 양성반응세포을 보였다. Bcl-2 mRNA에 대한 in situ hybridization에서는 성장난포와 퇴화난포의 기질세포 (interstitial cell), 난포막세포 (theca cell)에서 양성반응이 나타났고 PMSG를 투여한 후 1일, 2일군의 성장난포의 기질세포, 난포막세포에서 강하게 발현되었다. Bax mRNA는 성장난포와 퇴화난포의 기질세포, 난포막세포, 과립막세포 (granulosa cell)에서 양성반응이 나타났고 PMSG를 투여한 후 4일, 5일군의 퇴화난포의 기질세포, 난포막세포와 과립막세포에서 발현되었으므로 bcl-2와 bax의 비율이 세포의 운명을 결정하는데 중요한 역할을 한다는 사실을 확인 할 수 있었다. Bcl-2는 난포의 성장과 퇴화시 기질세포, 난포막세포에서 발현되어 난포의 퇴화를 억제하는 역할을 하며 bax는 퇴화하는 난포의 기질세포, 난포막세포와 과립막세포에서 발현되어 난포의 퇴화를 유도한다고 생각되었다. 따라서, bcl-2와 bax는 기질세포와 난포막세포에서 주로 합성되어 난포의 성장과 퇴화를 조절한다고 할 수 있다. In the mammalian ovary, follicular atresia occurs through apoptosis. Apoptosis is known as the physiological cell death, which is regulated by bcl-2 gene family. In the bcl-2 gene family, bcl-2/bcl-xLong is known as an inhibitor of apoptosis, whereas bax/bcl-xShort is known as an inducer of apoptosis. We thought that these genes are associated with follicular development and atresia. Therefore, the present study used a in situ hybridization to examine the expression of bcl-2 and bax mRNAs during ovarian follicular development and atresia induced by PMSG(15 IU) treatment in the immature rat ovary. Morphological changes were occurred with the manner of five days periodicity after PMSG treatment. One or two days after PMSG treatment, ovaries had growing follicles with antrum and healthy granulosa cells. Three days after treatment, some degenerating follicles which had thinner granulosa cell layers than growing follicles were observed. At day four and five after treatment, degenerating follicles which have pyknotic nuclei and thin distorted granulosa cell layers appeared. These atretic follicles showed positive reaction with in situ DNA end labelling which indicates apoptotic changes. This study showed that bcl-2 mRNA was expressed in the theca and interstitial cells. Growing follicles of one or two days have showed stronger bcl-2 mRNA signals than atretic follicles of four or five days in these cells. Bax mRNA was expressed in the theca cells, interstitial cells, and granulosa cells. Atretic follicles of four or five days showed stronger bax mRNA signals than growing follicles of one or two days in these cells. Expression of bcl-2 mRNA was increased in growing follicles while decreased in atretic follicles. In contrast, expression of bax mRNA was increased in atretic follicles while decreased in growing follicles. Therefore, we confirmed that follicular development and atresia were affected by the change in the ratio of bcl-2 and bax mRNAs. According to these data, we proposed that these two genes are associated with follicular development and atresia.

암세포 Bcl-2 family 유전자 군의 DNA 메틸화 연구

강영섭 ( Young Suep Kang ),이선영 ( Sun Young Lee ),정상근 ( Sang Gun Jung ),한지유 ( Ji You Han ),고정재 ( Jeong Jae Ko ),배지현 ( Jee Hyeon Bae ),나영정 ( Young Junh Na ),이찬 ( Chan Lee ),목정은 ( Jung Un Mock ),김승조 ( Sung 대한산부인과학회 2007 Obstetrics & Gynecology Science Vol.50 No.7

목적: 본 연구는 암세포주와 난소암 조직에서 세포예정사의 핵심 조절 단백질인 Bcl-2 family 유전자의 DNA 메틸화 여부를 암과의 관련성을 밝히는데 그 목적이 있다. 연구 방법: 자궁경부암 세포주인 HeLa, CaSki 그리고 만성골수성 혈액암 세포주인 K562 세포에서 genomic DNA를 추출하여 sodium bisulfite를 통한 cytosine 염기를 uracil로 치환하였다. 치환된 염기서열은 methylation과 unmethylation을 특이적으로 확인할 수 있는 primer를 이용하여 MSP (Methylation Specific PCR)을 수행한 후 암세포에서 Bcl-2 family 유전자의 DNA 메틸화 여부를 판별하였다. 결과: 각각의 세포주에서 antiapoptotic Bcl-2 family 유전자 군인 Mcl-1과 Bcl-2 유전자는 DNA 메틸화가 이루어지지 않은 것으로 관찰되었으며, proapoptotic Bcl-2 family 구성원인 Harakiri 유전자는 DNA 메틸화가 이루어진 것을 확인하였다. 반면, 다른 proapoptotic Bcl-2 family 유전자인 Noxa 유전자는 DNA 메틸화가 이루어지지 않은 것으로 관찰되었다. 또한 난소암 조직에서의 DNA 메틸화 여부를 살펴본 결과 Mcl-1과 Noxa는 세포주에서와 같은 결과를 보였고, Harakiri 유전자의 경우 hypomethylation으로 관찰되었다. 결론: 본 연구는 암세포주와 난소암 조직에서 proapoptotic Bcl-2 family 유전자인 Harakiri와 Noxa의 경우, 유전자 특이적인 DNA의 메틸화가 나타났으며 antiapoptotic 유전자는 DNA의 메틸화가 나타나지 않았다. 이로써 Bcl-2 유전자군과 암세포의 세포사멸을 억제하는 기작을 DNA의 메틸화를 통한 특이 유전자 발현 억제에 의해서 이루어질 수 있다고 사료된다. Objective: Promoter methylation of Bcl-2 family genes in cancer cells were studied to verify possible correlation between DNA methylation pattern of Bcl-2 family members and cancer. Methods: The genomic DNAs were extracted from different cancer cell lines, HeLa, CaSki and K562, and ovarian cancer tissue from patients. The cytosine residues were converted to uracil by sodium bisulfite treatment. MSP (methylation specific PCR) was performed to determine the methylation status of Bcl-2, Mcl-1, Noxa, and Harakiri promoters. Using primers that distinguish methylated DNA from unmethylated DNA after bisulfite modification of DNA, MSP was conducted to observe the methylation pattern of Bcl-2 family genes in different cancer cells. Results: The promoter regions of Bcl-2 family genes including Mcl-1, Bcl-2, and Noxa were not methylated in cancer cells, whereas the proapoptotic Bcl-2 family gene Harakiri was detected as methylated in the cancer cell lines and hypomethylated in the ovarian cancer tissue. Conclusion: The present study demonstrated the differential methylation profiles of Bcl-2 family genes in cancerous cells, which suggests a possible connection between the methylation pattern of some of Bcl-2 family genes and ovarian cancer.

KGN(난소과립세포)에서 BCL2L10 단백질의 세포사멸 유도 기능 연구

김재홍,이경아,배지현 한국발생생물학회 2011 발생과 생식 Vol.15 No.2

BCL-2 family 단백질들은 세포사멸 신호전달 체계에서 중추적인 역할을 수행하는 것으로 알려져 있으며, BCL2L10 단백질은 그 중 하나로 세포의 사멸과 생존을 조절하는 것으로 알려져 있다. 특이하게도 BCL2L10 단백질은 세포 또는 조직 특이적으로 서로 상반되는 친 세포사멸 또는 항 세포사멸 효과가 각각 보고되어 있다. 현재까지 난소세포에서의 BCL2L10의 발현 여부 및 기능은 알려져 있지 않다. 따라서 본 연구에서 인간 난소 과립세포주인 KGN 세포에서의 BCL2L10 단백질의 발현 여부를 확인한 결과, 상당한 양의 단백질이 발현함을 확인할 수 있었으며, 또한 세포사멸 효과를 확인하기 위해서 BCL2L10 단백질을 dose-dependent하게 과발현시킨 후 세포의 생존에 미치는 영향을 분석한 결과, BCL2L10은 KGN 세포에서 과발현 시 세포사멸을 유도함을 관찰하였다. BCL2L10 단백질을 과발현 시 Caspase 9와 3를 활성화 하였으며, 면역염색법을 통해서 BCL2L10 단백질이 미토콘드리아에 위치하는 것을 확인하였다. 또한 BCL2L10 단백질의 과발현에 의해 미토콘드리아에서 cytochrome c가 세포질로 분비되는 것을 확인하였다. 이상의 결과로서 본 연구는 BCL2L10의 과발현이 KGN 세포에서 세포사멸을 유도하고, 또한 미토콘드리아에 위치하여 세포질로 cytochrome c를 분비하여 Caspase 9와 3을 활성화 시키는 메커니즘으로 세포사멸을 유도함을 확인하였다. 이러한 연구결과는 BCL2L10 단백질이 인간 난소과립세포의 생존과 사멸을 조절하는 인자임을 최초로 규명한 것으로서, 추후 난소에서의 BCL2L10 단백질의 생리적인 기능 및 신호 조절 연구의 기반 데이터로서 그 의의가 있으며 활용될 수 있다.

유방암 아형에 따른 예후 인자로서의 Bcl-2

이주영,김현아,김은규,양회민,김관일,이종인,고재수,고은영,문난모,김민석,백남선,노우철 한국유방암학회 2011 Journal of breast cancer Vol.14 No.-

Purpose: B-cell lymphoma (bcl)-2 is an anti-apoptotic gene, and it is a poor prognostic factor in various malignant tumors. However, the prognostic significance of bcl-2 expression in breast cancer remains controversial. We investigated the prognostic significance of bcl-2 according to cancer molecular subtype. Methods: We analyzed 411 patients with primary invasive breast cancer who underwent surgery at our institution between 1999 and 2001. The subtypes were classified as luminal (estrogen receptor [ER]+ and/or progesterone receptor [PR]+, irrespective of human epidermal factor receptor 2 [HER2]), triple-negative (ER-, PR-, and HER2-), or HER2 (ER- ,PR-, and HER2+). Results: A total of 236 (57.4%) cases were positive for bcl-2, and bcl-2 expression was significantly associated with earlier stage, lower grade, expression of hormone receptor positivity, and HER2 negativity. No difference in disease-free survival (DFS) was observed based on bcl-2 expression. However, the prognostic significance of bcl-2 varied with subtype; bcl-2 was not a prognosticator in patients with the luminal and HER2 subtypes. However, patients with bcl-2(+) tumors of the triple-negative subtype showed significantly worse DFS than those with bcl-2(-) tumors (p=0.048). In a multivariate analysis, bcl-2 expression remained a significant predictor of recurrence in patients with the triple-negative subtype (hazard ratio, 3.26; 95% confidence interval, 1.40-7.59; p=0.006). Conclusion: The prognostic significance of bcl-2 varied with molecular subtype; bcl-2 expression was a poor prognosticator in patients with the triple-negtive subtype, but not in those with the luminal and HER2 subtypes.

유방의 침윤성 유관암에서 cyclin D1, bcl-2의 발현: 그 연관관계와 임상적 의미

홍영기,백승삼,정민성,윤호성 한국유방암학회 2008 Journal of breast cancer Vol.11 No.4

Purpose: Cyclin D1 and bcl-2 are involved in cell proliferation and apoptosis in tumor development and are commonly expressed in breast cancer. But there are few clinical reports on the correlation between cyclin D1 and bcl-2 expression. This study was designed to analyze the correlations of cyclin D1 and bcl-2 and their clinical implications in breast cancer. Methods: Immunohistochemical expression of cyclin D1 and bcl-2 were studied in 342 infiltrative ductal carcinoma cases and were compared with clinicopathologic parameters such as age, tumor size, histologic grade, lymph node status, p53, c-erbB2 and hormone receptors. Results: Cyclin D1 expression was found in 86 of 342 cases (25.1%). Bcl-2 was positive in 227 of 342 cases (66.4%). Bcl-2 overexpression was associated with the high expression of cyclin D1 (p=0.001). Correlation was detected between both cyclin D1 and bcl-2 and hormone receptor positivity (p<0.001). There was a reverse correlation between bcl-2 and histologic grade, p53, c-erbB2. And the bcl-2 overexpression group showed better disease free survival rates at 3-year follow up. Conclusion: Higher expression of cyclin D1 was associated with bcl-2 overexpression. Positive estrogen receptor expression was associated with high cyclin D1 and bcl-2 expression. Bcl-2 tends to correlate with a positive clinical outcome.