Expression of the survivin-2B splice variant related to the progression of colorectal carcinoma

Gyu-Seok Cho,Tae Sung Ahn,Dongjun Jeong,Jae-Jun Kim,Chang-Jin Kim,Hyun-Deuk Cho,Dong-Kook Park,Moo-Jun Baek 대한외과학회 2011 Annals of Surgical Treatment and Research(ASRT) Vol.80 No.6

Purpose: Recently, two alternatively spliced survivin variants, survivin-ΔEx3 and survivin-2B, were identified in a single copy of the survivin gene. It has been reported that the expressions of survivin splice variants significantly correlates with the clinical results in many types of human carcinoma. We investigated the transcription levels of survivin and its splice variants in human colorectal carcinomas, and analyzed correlations between survivin expression levels and clinicopathologic features. Methods: We used Western blot and real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) to analyze the protein and mRNA expression levels of survivin variants in 51 colorectal carcinomas. The quantitative RT-PCR was performed using primer pairs specific for survivin and each of its splice variants, then normalized for the gene that encodes glyceraldehydes-3-phosphate dehydrogenase. Results: In Western blotting, the protein levels of survivin were higher in the tumor tissue than in normal tissue. The expression of survivin, survivin-2B and survivin-ΔEx3 mRNA was present in 96%, 64.7%, and 82.4% of the samples, respectively. When the pathologic parameters were compared, colorectal cancers of advanced pT stages showed significant decrease in survivin-2B mRNA expression by the quantitative RT-PCR (P < 0.001). Conclusion: The decreased expression of survivin-2B might be related to tumor progression in colorectal cancers. This finding indicates that alternatively spliced variants of survivin may be involved in refining the functions of survivin during tumor progression.

Change in Expression of Survivin Caused by Using Oxaliplatin in HCT116 Colon Cancer Cells

손원준,이정원,박동국 대한대장항문학회 2010 Annals of Coloproctolgy Vol.26 No.4

Purpose: Oxaliplatin is a third-generation platinum compound, and it has no nephrotoxicity and has reduced bone marrow toxicity. Cancer cells that are resistant to cisplatin are sensitive to oxaliplatin. Oxaliplatin is used widely for the treatment of colon cancers. Recently, oxaliplatin was reported to inhibit the expression of survivin, which protects cell apoptosis. However, there are no reports on the expressions of survivin variants and the changes in intracellular localization of survivin in cancer cells. We studied the expression of survivin caused by oxaliplatin in HCT116 colon cancer cells, and we observed the localization of survivin in the mitotic phase. Methods: We treated the HCT116 colon cancer cells with 2.0 μM of oxaliplatin, and we studied the expressions of survivin protein, and survivin mRNA variants, as well as the changes in intracellular localization, by using the Western blot method,RT-PCR, immunocytochemistry, and flowcytometry. Results: Oxaliplatin inhibits the expression of the survivin protein and survivin mRNA in HCT116 colon cancer cells. The expression of the survivin-2B variants, which have no antiapoptotic activity but control cell mitosis by localization on a microtubule, is reduced continuously 2 days after treatment with oxaliplatin. In immunocytochemistry, expression of survivin in the cytoplasm is reduced and especially is not expressed in microtubules and contractile rings. Conclusion: One of the mechanisms of oxaliplatin is to inhibit the expression of and to change the localization of survivin. Based on these results, we suggest that changes in the expression of survivin variants and in their localization are two effects of oxaliplatin.

A novel strategy to promote liver regeneration: utilization of secretome obtained from survivin-overexpressing adipose-derived stem cells

Cho-Hee Kim,Ok-Hee Kim,Jung Hyun Park,Say-June Kim 대한외과학회 2021 Annals of Surgical Treatment and Research(ASRT) Vol.101 No.6

Purpose: Survivin is a typical antiapoptotic protein. It is copiously expressed during human fetal development but is infrequently present in adult tissues. In this experiment, we researched the treatment effect of the secretome that adipose-derived stem cells (ASCs) transfected with survivin. Methods: First of all, we generated survivin-overexpressing ASCs transfected with a plasmid comprising a gene encoding survivin. The secreted substances released from survivin-overexpressing ASCs (survivin-secretome) were collected, and were determined their in vitro and in vivo therapeutic potential, especially in the model of liver impairment. Results: In vitro, the survivin-secretome significantly increased cell viability and promoted the expression of proliferation-related markers (proliferating cell nuclear antigen [PCNA], phospho-signal transducer and activator of transcription 3 (p-STAT3), hepatocyte growth factor [HGF], vascular endothelial growth factor [VEGF]) and anti-apoptosis-related markers (myeloid cell leukemia-1 [Mcl-1] and survivin) (P < 0.05). In vivo using 70% hepatectomy mice, the survivin-secretome group exhibited the lowest serum levels of interleukin-6, tumor necrosis factor-α (P < 0.05). The serum levels of liver transaminases (alanine aminotransferase and aspartate aminotransferase) were also the lowest in the survivin-secretome group (P < 0.05). The survivin-secretome group also exhibited the highest liver regeneration on the 7th day after 70% partial hepatectomy (P < 0.05). In the subsequent liver specimen analysis, the specimens of survivin-secretome exhibited the highest expression of p-STAT3, HGF, VEGF, PCNA, and Mcl-1 and the lowest expression of bcl-2-like protein 4 (P < 0.05). Conclusion: Taken together, secretome secreted by survivin-overexpressing ASCs could be an effective way to improve liver regeneration and repair for liver injury treatment.

대장암 환자에서 Survivin mRNA의 발현과 간 전이와의 연관성

최종순,장희경 고신대학교의과대학 2008 고신대학교 의과대학 학술지 Vol.23 No.1

연구배경 및 목적 Survivin은 새로이 고사(아포프토시스)-억제 유전자군에 추가된 유전자이다. Survivin 발현이 대장암에서 보고는 되고 있으나, 상이한 결과들로 논란이 많으며, 임상적 의의에 대해서도 불명확하다. 더욱이 이 유전자는 암 치료나 예방적 항암치료의 표적으로의 가능성이 제기되고 있다. 대장암 survivin 유전자의 발현을 조사하고 임상 및 예후 인자들과의 관련성을 분석하여 대장암의 발생과 임상 측면에서이들 유전자의 역할을 이해하고 예방과 치료에 이용할 수 있는 지를 조사하고자 하였다. 대상 및 방법 다양한 병기의 37예의 신선한 대장암 수술 조직을 대상으로 하였다. 암 조직과 주위 비종양성 조직에서 전체 mRNA를 분리하여 역전사-연쇄중합효소반응으로 survivin mRNA를 검출하였다. PCR 산물을 확인하기 위하여 아클로닝후에 DNA 염기 분석을 시행하였다. 결 과 대장에서 Survivin의 발현율은 대장암 조직에서는 94.6%였고, 비종양성 조직에서는 51.4%으로, 대장암 조직에서 survivin의 발현이 높았으나, 비종양성 조직에서의 경계성 영역의 유의한 차이가 인정되었다(0.05<p=0.065< 0.1). 연령, 성별, 침윤 깊이, 림프절 전이, 혈관과 신경 침범, 간으로의 전이 등의 임상 및 병리학적 인자들과 관련성은 없었다. 결 론 survivin은 대장암에서 암특이적으로 발현되지 않았다 (0.05<p=0.065< 0.1). 대장암의 치료나 chemoprevention에서 SSX(synovial sarcoma on X chromosome gene)와 survivin의 이용은 가능하나 제한적일 것으로 생각된다. SSX와 survivin 발현의 상관성이 인정되지 않으므로, 대장암에서 SSX은 survivin-관련 항아포토시스 경로와는 무관할 것으로 생각된다. Background : Survivin is a novel member of the inhibitors of apoptosis proteins (IAP) gene family. Although survivin expression has been reported in colon cancer, the results are still controversial and its clinical significanceincluding chemoprevention remains unclear. Some reports described thegene as a potential target in anticancer or chemopreventive therapy. To facilitate understanding of survivin in the colon cancer, the clinicopathological significance was investigated with the expression of survivin. Methods : Surgical specimen were obtained from a total 37 cases of consecutive patients with various stages of colorectal carcinoma who had undergone a resection. To determine survivin mRNA in colorectal carcinomas and adjacent normal colorectal tissue samples, total RNA was isolated from each of the samples after lysis in quanidinium isothiocyanate and phenol extraction and reverse-transcription polymerase chain reaction was done. The PCR products were confirmed by DNA sequencing after subcloning. Results : The positive rate of survivin in cancer tissue was 94.6%(35/37) and that of non-neoplastic colorectal tissue was 51.4%(19/37). S urvivin expression t ended to i ncrease in c ancer tissue, but h as s tatistically m arginal significance(0.05<p=0.065< 0.1). The correlation of survivin expressions to age, gender, invasion depth, lymph node metastasis, vessel and perineural invasions, and liver metastasis was not found. Conclusions : The expression of survivin appears not to be eligible to differentiate malignant from benign lesions in colon, even though it has marginal significance(0.05<p=0.065< 0.1). The role of survivin-associated apoptosis may be unnoticeable, in the light of insignificant correlation of gene expression to the other prognostic factors in colon cancer.

전립선비대증 환자에서 5알파환원효소 억제제 투여가 Survivin과 Bcl-2의 발현에 미치는 영향

차우헌,장태정,이경섭 대한비뇨의학회 2008 Investigative and Clinical Urology Vol.49 No.3

Purpose: A 5-alpha-reductase inhibitor(5αRI) can induce apoptosis and decrease the prostatic volume in patients with benign prostatic hyperplasia (BPH). In this study we assessed the expression of survivin and bcl-2 in the epithelium of BPH patients treated with finasteride. Materials and Methods: Group 1 consisted of 39 patients who underwent transurethral resection of the prostate(TURP) without medication and Group 2 consisted of 31 patients who underwent TURP and were treated with finasteride for more than three months. The mean age of both groups of patients was 73.03±7.02 years and 74.71±5.99 years, respectively. Immunohistochemical staining for survivin, bcl-2 and ki-67 was performed in prostatic tissues. The percentage of cells that expressed survivin and bcl-2 were classified into four categories based on the staining intensity. The expression of ki-67 in nuclei using 10 random cells per specimen was obtained. The relationship between 5αRI and the expression of survivin, bcl-2 and ki-67 was analyzed. Results: The total mean prostate volume of group 1 patients and group 2 patients was 45.51ml and 37.23ml, respectively (p<0.001) and the mean serum total prostate-specific antigen(PSA) level of group 1 patients and group 2 patients was 5.09ng/ml and 3.75ng/ml, respectively(p=0.105). Decreased expression of survivin and bcl-2 in specimens from group 2 patients was observed as compared to the level of expression in group 1 patients(p<0.001, p=0.001). Expression of ki-67 determined in samples from both groups was not significantly different(p=0.345). Conclusions: We suggest that finasteride may induce apoptosis of prostatic epithelial cells in BPH patients by reducing the expression of survivin and bcl-2. These findings may indicate a reduction of prostatic volume. (Korean J Urol 2008;49:242-247) Purpose: A 5-alpha-reductase inhibitor(5αRI) can induce apoptosis and decrease the prostatic volume in patients with benign prostatic hyperplasia (BPH). In this study we assessed the expression of survivin and bcl-2 in the epithelium of BPH patients treated with finasteride. Materials and Methods: Group 1 consisted of 39 patients who underwent transurethral resection of the prostate(TURP) without medication and Group 2 consisted of 31 patients who underwent TURP and were treated with finasteride for more than three months. The mean age of both groups of patients was 73.03±7.02 years and 74.71±5.99 years, respectively. Immunohistochemical staining for survivin, bcl-2 and ki-67 was performed in prostatic tissues. The percentage of cells that expressed survivin and bcl-2 were classified into four categories based on the staining intensity. The expression of ki-67 in nuclei using 10 random cells per specimen was obtained. The relationship between 5αRI and the expression of survivin, bcl-2 and ki-67 was analyzed. Results: The total mean prostate volume of group 1 patients and group 2 patients was 45.51ml and 37.23ml, respectively (p<0.001) and the mean serum total prostate-specific antigen(PSA) level of group 1 patients and group 2 patients was 5.09ng/ml and 3.75ng/ml, respectively(p=0.105). Decreased expression of survivin and bcl-2 in specimens from group 2 patients was observed as compared to the level of expression in group 1 patients(p<0.001, p=0.001). Expression of ki-67 determined in samples from both groups was not significantly different(p=0.345). Conclusions: We suggest that finasteride may induce apoptosis of prostatic epithelial cells in BPH patients by reducing the expression of survivin and bcl-2. These findings may indicate a reduction of prostatic volume. (Korean J Urol 2008;49:242-247)

골반강 외 자궁내막증에서 Survivin 발현

김영아 ( Young Ah Kim ),김한성 ( Han Seong Kim ),권혜성 ( Hye Seong Kwon ),신상현 ( Sang Hyun Shin ),장두영 ( Doo Young Chang ),최형민 ( Hyung Min Choi ),전명권 ( Myung Kwon Jeon ) 대한산부인과학회 2008 Obstetrics & Gynecology Science Vol.51 No.12

목적: 골반강 외 자궁내막증 조직에서 새로운 세포고사 저해 단백질인 survivin의 발현을 정상자궁내막과 난소 자궁내막종으로 비교 조사함으로써 골반강 외 자궁내막증의 병태생리에 세포고사가 어떻게 작용하는지 알아보고자 하였다. 방법: 본 연구는 수술 후 조직검사에서 골반강 외 자궁내막증으로 진단된 14예를 복강경으로 진단되어 제거한 난소 자궁내막종조직 13예와 자궁내막증이 없고 자궁근종으로 자궁을 적출한 정상자궁내막 34예를 대조군으로 하여 연구를 시행하였다. 면역조직학적 염색은 survivin은 rabbit antihuman polyclonal antibody를 이용하여 시행하였다. 결과: 골반강 외 자궁내막증에서 survivin의 발현은 선세포와 기질세포에서 핵내 발현은 정상자궁내막보다 증가하며 통계학적 차이를 보였다. 그러나 세포질 내 발현은 오히려 정상자궁내막이 통계학적 차이를 보이며 증가하였다. 난소 자궁내막종에서 survivin의 발현은 선세포와 기질세포에서 핵내 발현은 정상자궁내막보다 의미 있게 증가하나, 선세포와 정상자궁내막의 증식기 비교에서는 통계학적 차이를 보이지 않았다. 세포질 내 발현은 정상자궁내막이 통계학적 차이를 보이며 증가하였다. 골반강 외 자궁내막증과 난소자궁내막증에서 survivin의 발현의 차이는 없었다. 결론: 골반강 외 자궁내막증 환자에서 세포고사를 저해하는 survivin의 발현이 증가하는 것은 survivin이 골반강 외 자궁내막증의 병태 생리와 관련이 있을 것으로 생각된다. Objective: To examine survivin expression in extrapelvic endometriosis. Methods: The study group consisted of 14 cases with extrapelvic endometriosis which were confirmed histologically. The control group (total, n=47) was divided into 2 groups. Group I included normal endometrium (n=34) obtained from hysterectomy specimens with myoma and without endometriosis. Group II included ovarian endometrioma (n=13) obtained from laparoscopy. Expression of survivin was immunohistochemically confirmed. Results: In extrapelvic endometriosis, the expression of nucleus in glandular epithelium and stromal cells were significantly stronger than normal endometrium. But cytoplasm expression of glandular epithelial cells and stromal cells in extrapelvic endometriosis showed statistically lower in comparison with normal endometrium. In ovarian endometrioma, the expression of nucleus in glandular epithelial cells and stromal cells was significantly stronger than normal endometrium. But the expression of nucleus in glandular epithelial cells with ovarian endometrioma was stronger than during proliferative phase but was not significant. Also cytoplasm expression of ovarian endometrioma was lower than normal endometrium. There was no difference in survivin expression between extrapelvic endometriosis and ovarian endometrioma. Conclusions: In extrapelvic endometriosis, survivin expression was stronger than normal endometrium except cytoplasm. Our findings suggest that increased survivin expression may contribute to survival of extrapelvic implants.

Experimental Research Article : The expression of survivin and its related genes in adipocyte-derived stem cell by demethylation

( Kwang Yoon ),( Young Soo Lim ),( Soo Bong Yu ),( Doo Sik Kim ),( Sie Jeong Ryu ),( Kyung Han Kim ),( Tae Ho Jang ),( Se Hwan Kim ) 대한마취과학회 2010 Korean Journal of Anesthesiology Vol.58 No.4

Background: Survivin is thought to contribute to stem cell maintenance partly by a hypomethylation mechanism. This study attempted to elucidate the signal transduction pathway of adipocyte-derived stem cells (ASCs) by using a demethylating agent, 5-aza-2`-deoxycytidine (ADC), to analyze the survivin, MEK/ERK, c-Myc and p53 gene expression. Methods: Demethylation in the ASCs was induced by 1 μM ADC treatment. RT-PCR for survivin mRNA was preformed, before and 24, 48 and 72 hours (hr) after ADC treatment. Western blotting analysis was performed for p53, survivin, unphosphorylated and phosphorylated (p)-MEK, and p-ERK. Immunohistochemistry for ERK and survivin was done to evaluate the localization of the proteins. Results: ADC inhibited the population growth of the ASCs and it increased the number of apoptotic cells 24, 48, and 72 hr after treatment. ADC treatment slightly decreased the expression of survivin mRNA after 48 hr and its level was restored after 72 hr of treatment. Otherwise, the level of survivin protein gradually increased up to 48 hr and it was decreased at 72 hr. The levels of p-MEK and p53 were increased time-dependently. c-Myc and p-ERK were elevated after ADC treatment and their highest levels were seen 48 hr after treatment. The ADC treatment increased the nuclear expression of ERK and survivin in the ASCs. Conclusions: The overexpression of p-MEK/ERK, p53, and c-Myc increased the survivin protein expression of the demethylated ASCs. These results suggest that demethylation could alter the expression of survivin, and p53, c-Myc and the MAPK (MEK/ERK) pathway might play a role in survivin`s regulation in ASCs. (Korean J Anesthesiol 2010; 58: 383-390)

Clinicopathologic Significance of Survivin Expression in Relation to CD133 Expression in Surgically Resected Stage II or III Colorectal Cancer

Wanlu Li,이미라,최은희,조미연 대한병리학회 2017 Journal of Pathology and Translational Medicine Vol.51 No.1

Background: Cancer stem cells have been investigated as new targets for colorectal cancer (CRC) treatment. We recently reported that CD133+ colon cancer cells showed chemoresistance to 5-fluorouracil through increased survivin expression and proposed the survivin inhibitor YM155 as an effective therapy for colon cancer in an in vitro study. Here, we investigate the relationship between survivin and CD133 expression in surgically resected CRC to identify whether the results obtained in our in vitro study are applicable to clinical samples. Methods: We performed immunohistochemical staining for survivin and CD133 in surgically resected tissue from 187 stage II or III CRC patients. We also comparatively analyzed apoptosis according to survivin and CD133 expression using terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling. Results: The results of the Mantel-Haenszel test established a linear association between nuclear survivin and CD133 expression (p = .018), although neither had prognostic significance, according to immunohistochemical expression level. No correlation was found between survivin expression and the following pathological parameters: invasion depth, lymph node metastasis, or histologic differentiation (p > .05). The mean apoptotic index in survivin+ and CD133+ tumors was higher than that in negative tumors: 5.116 ± 4.894 in survivin+ versus 4.103 ± 3.691 in survivin– (p = .044); 5.165 ± 4.961 in CD133+ versus 4.231 ± 3.812 in CD133– (p = .034). Conclusions: As observed in our in vitro study, survivin expression is significantly related to CD133 expression. Survivin may be considered as a new therapeutic target for chemoresistant CRC.

The Expression of Cyclooxygenase-2 and Survivin in Urinary Bladder Transitional Cell Carcinoma

장태정,이경섭 대한병리학회 2009 Journal of Pathology and Translational Medicine Vol.43 No.3

Background : The aim of this study was to investigate the expressions of cyclooxygenase-2 (COX-2) and survivin in bladder transitional cell carcinoma (TCC) that has different clinicopathologic characteristics, and we also wanted to determine if a relation exists between the COX-2 and survivin expressions. Methods : The expressions of COX-2 and survivin were investigated in 80 bladder TCCs by performing immunohistochemistry. Results : The normal bladder mucosa did not express COX-2 and survivin. COX-2 immunopositivity and cytoplasmic survivin immunopositivity were seen in 48% and 30% of bladder tumors, respectively. The expressions of COX-2 and survivin were closely related to the differentiation, depth and recurrence of bladder TCC, and there was a significant correlation in topographic distribution of COX-2 and survivin immunopositivity. In addition, COX-2 and survivin were predominantly expressed at the invasive front of tumors. Conclusions : This data suggest that COX-2 and survivin may be involved in the progression of bladder TCC, and there is a close correlation between the expressions of COX-2 and survivin.

후두편평세포암에서 Vascular Endothelial GrowthFactor(VEGF)와 Survivin의 발현

김효진,김기현,장영도,조성호,고윤우,김동욱,이병돈,장혁순 대한이비인후과학회 2006 대한이비인후과학회지 두경부외과학 Vol.49 No.12

Background and Objectives:Angiogenesis and alteration of apoptosis are hypothesized to be the mechanisms of the growthof tumor. Vascular endothelial growth factor (VEGF) is considered to be the most primary factor prompting the angiogenesis intumor tissue, which also holds the central position in the course of formation and metastasis of tumor and regulates expression ofsurvivin. The survivin protein is the member of inhibitor of apoptosis protein (IAP) family which inhibits apoptosis. Recently,several authors reported survivin and VEGF expression was found in various cancer tissue and they are suggest to play on theimportant role in cancer development. The purpose of this study is to evaluate the expression of VEGF and survivin and toinvestigate their correlation with the clinical stage, nodal involvement, and histologic grade in the squamous cell carcinoma oflarynx. Subjects and Method:Immunohistochemical staining for the paraffin sections by using a polyclonal antibody forVEGF and survivin by the standard avidin-biotin-peroxidase technique was performed in 19 cases with the squamouse cellcarcinoma of larynx. The relationships between the expression of VEGF and survivin and clinicopathological characteristicswere analyzed. Results:Immunohistochemical analysis showed the expression of VEGF in 15 of 19 cases (78.9%) and survivinin 19 of 19 cases (100%). There was no correlation between the expression of VEGF and survivin and the clinical stage, nodalinvolvement, and histologic grade in the squamous cell carcinoma of larynx (p>0.05). But VEGF results in an increased expressionof survivin (p<0.05). Conclusion:Further study will be needed to understand the relationships between the VEGFand survivin and squamous cell carcinoma of larynx. (Korean J Otolaryngol 2006;49:1188-93)