알코올성 간 손상 조직에서 TGF-β1와 c-Myc, Erb-B2, Thymosin-β4 유전자 발현 융합 연구

김진수,최상기 한국융합학회 2018 한국융합학회논문지 Vol.9 No.5

본 연구는 25% 에탄올에 손상된 간조직에서 TGF-β 1와 c-Myc, Erb-B2, Thymosin-β 4 유전자의 발현을 알아 보고 자 실시 하였다. 실험군은 2군으로 나누어 25% 에탄올로 간 손상을 유발한 실험군과 정제수를 투여한 대조군으로 나누어 실험하였다. 검사 결과는 25% 에탄올를 투여 했던 실험군은 대조군에 비하여 TGF-β 1, c-Myc 및 Thymosin-β 4 유전자의 발현 증가를 알 수 있었으며 Erb-B2 유전자는 뚜렷한 발현을 알 수 없었다. 또한 손상된 간 조직에서 헤마톡실린 에오진 염색을 통한 세포 손상을 관찰 할 수 있었다. 결론적으로 기존 임상에서 간 기능 관련 질병 예방과 질환 판정 시 혈청학적, 조직학적 검사 외에 TGF-β 1, c-Myc 및 Thymosin-β 4의 분자 진단 기법에 의한 유전자 발현 상태를 융합 검사함으로써 간 질환 판정의 보조 자료로 활용 될 수 있을 것으로 사료 된다. This study has been conducted to see the expression of TGF- β1, c-Myc, Erb-B2 and Thymosin- β4 genes in ethanol - damaged liver tissues. Experimental groups were divided into 2 groups, one where damaged liver was caused by 25% ethanol and normal group administered with purified water. Results of test showed the expression of TGF- β1, c-Myc, and Thymosin- β4 genes was higher in the experimental group treated with 25% ethanol than in the normal group. Erb-B2 gene was not expressed clearly. Thus, it is considered that we can expect to utilize TGF- β1, c-Myc 및 Thymosin- β4 as auxiliary data and find clinical meanings of diagnosis on hepatic diseases, In addition to serologic and histological examination by convergence examining the gene expression status by molecular diagnostic techniques in liver-related disease prevention and diagnosis through results of this study.

섬유화 진행 간경변 조직 모델에서 Thymosin β4와 C-myc mRNA 융합 발현 연구

김진수,박운규 한국융합학회 2019 한국융합학회논문지 Vol.10 No.6

The propose of this study has been conducted to examine expression of c-Myc and Thymosin-β4 in liver cirrhosis model from liver fibrosis and For the method of study, the experiment was conducted in 2 groups; liver cirrhosis model experiment group due to liver fibrosis and control group with distilled water. This study outcome showed that liver cirrhosis model experiment group had significantly higher expression of c-Myc and Thymosin-β4. with changes to hepatic tissue of special staining and electron microscopy. In conclusion, in clinical tests regarding liver function, molecular evaluation of c-Myc and Thymosin-β4 and their expression along with serological change and histological assessment can be utilized as a reference for diagnosing liver disease for prevention and diagnosis of the disease, Based on this research in the future, we will carry out an in-depth study by adding the types of experimental groups and related genes. 이 연구의 목적은 간 섬유화로 인한 간경변 모델에서 c-Myc과, Thymosin-β4 유전자의 발현을 알아 보고자 실시하였으며 연구 방법은 2군으로 나누어 섬유화로 인한 간경변 모델 실험군과 증류수를 이용한 대조군으로 분류 한 후 실험을 하였다. 본 연구를 통한 결과는 간경변 모델 실험군은 대조군에 비하여 c-Myc 과 Thymosin-β4 의 발현 이 증가 되는 것을 뚜렷하게 알 수 있었고, 전자현미경적 검경 외에 여러 특수 염색을 통하여 간조직의 변화를 관찰 할 수 있었다. 결론적으로 기존 간기능 검사 관련 임상검사에서 질환 예방과 질환 판정 시 혈청학적 변화와 조직학적인 검사와 더불어 c-Myc 과 Thymosin-β4의 분자 기법을 통한 유전자의 발현 상태를 확인함으로써 간 질환 판정에 기여 할 수 있을 것으로 사료 되며. 향후 이 연구를 기반으로 실험군의 종류와 관련 유전자를 추가 하여 심도 있는 연구를 진행할 예정이다.

자궁경부암에서 Annexin 1 및 Thymosin β4의 발현에 관한 연구

이유선 ( Yu Sun Lee ),정혜원 ( Hye Won Chung ),문혜성 ( Hye Sung Moon ) 대한산부인과학회 2008 Obstetrics & Gynecology Science Vol.51 No.3

목적: Annexin I은 암세포의 침윤 및 맥관형성을 자극하는 인자이고, thymosin β4는 암의 전이와 맥관형성에 관여하는 인자로서, 자궁경부암에서 annexin I 및 thymosin β4의 발현에 관하여 알아보고자 하였다. 방법: 33예의 침윤성 자궁경부암 환자와 10예의 자궁경부상피내종양 환자, 10예의 정상자궁경부 조직에서 QC-PCR과 western blot analysis를 이용하여 annxin I, thymosin β4 mRNA와 protein의 발현을 측정하였고, SiHa 세포주에 thymosin β4 peptide를 처리한 후 처리 전후의 annexin I의 발현 변화를 관찰하였다. 결과: 자궁경부암에서 thymosin β4와 annexin I mRNA의 발현은 정상 자궁경부, 자궁경부상피내종양에 비해 통계적으로 유의하게 높았으며, thymosin β4와 annexin I 단백질도 정상자궁경부보다 자궁경부암에서 높게 발현되었다. Thymosin β4와 annexin I mRNA의 발현은 다른 예후인자와 유의한 상관관계가 없었으나, thymosin β4와 annexin I 단백질의 경우 임상병기가 진행될수록 발현이 증가하는 양상을 보였다. Thymosin β4 peptide를 처리한 SiHa 세포에서는 처리하지 않은 세포보다 annexin I의 발현이 약간 증가하는 소견을 보였다. 결론: Annexin I와 thymosin β4의 발현은 침윤성자궁경부암에서 증가하고, 서로 상호작용을 일으켜 자궁경부암의 진행에 관여할 것으로 생각된다. 이 두 인자는 자궁경부암에서 진행 및 전이 촉진에 대한 생물학적인 지표로서의 가능성이 제시될 수 있을 것으로 사료된다. Objective: The aim of this study was to compare the expression of annexin I and thymosin β4 in invasive cervical cancer including normal cervix and CIN. Methods: In Ewha Womans University Mokdong Hospital, normal cervical tissues were obtained from healthy women (n=10), from patients with cervical intraepithelial neoplasia (CIN, n=10) and from patients with cervical cancer (n=33). The expressions of annexin I and thymosin β4 mRNA and protein were examined by quantitative competitive-PCR and by western blot analysis. The expressions of annexin I and thymosin β4 protein were measured by western blot analysis with thymosin β4 peptides non treated and treated SiHa cells. Results: The expression of thymosin β4 mRNA and protein in cervical cancer were higher than that in normal cervix (p<0.05). The expression of annexin I mRNA and protein were higher than that in normal cervix and CIN (p<0.05). Thymosin β4 and annexin I mRNA expressions were not significantly correlated with cervical cancer stage, or size of the tumor (p>0.05). But thymosin β4 and annexin I protein expressions were increased according to the cancer stage. The expression of annexin I was slightly higher in thymosin β4 treated SiHa cells than that in nontreated SiHa cells. Conclusions: Our results suggest that overexpression of thymosin β4 and annexin I may play roles in progression of invasive cervical cancer. Thymosin β4 upregulates expression of annexin I in invasive cervical cancer. Therefore, thymosin β4 and annexin I may be biological markers in detecting the progression of invasive cervical cancer, and their interaction is important in invasive cervical cancer.

상피성 난소암에서 혈관신생인자로서 Thymosin β4의 발현에 관한 연구

김아리 ( A Ri Kim ),정혜원 ( Hye Won Chung ),김종일 ( Jong Il Kim ),전선희 ( Sun Hee Chun ),문혜성 ( Hye Sung Moon ) 대한산부인과학회 2008 Obstetrics & Gynecology Science Vol.51 No.5

목적: 본 연구는 상피성 난소암에서 혈관신생인자로서 Thymosin β4의 발현 및 그 의미를 알아보고자 하였다. 방법: 18예의 정상 난소조직과 27예의 상피성 난소암 조직을 대상으로 하였다. Thymosin β4 mRNA의 발현은 quantatitive competitive PCR을 사용하여 측정하였다. 혈관 신생인자로서 thymosin β4의 발현과 함께 vascular endothelail growth factor (VEGF), angiopoietin-1, 2 (Ang-1,2)의 발현도 함께 측정하였다. 결과: Thymosin β4 mRNA의 발현은 상피성 난소암에서 정상 난소조직에 비해 통계적으로 유의하게 높게 발현되었다 (p<0.05). Thymosin β4 mRNA의 발현은 상피성 난소암에 있어 병기, 조직학적 분류, 수술 전 혈청 CA 125수치 및 림프절 전이 여부와 유의한 상관관계는 없었으나, VEGF, Ang-1, 2의 mRNA 발현과는 유의한 상관관계를 보였다 (p<0.05). 결론: Thymosin β4 mRNA의 과발현은 난소 종양에서 혈관 신생에 중요한 역할을 할 것으로 예측되며, 상피성 난소암을 진단하는 생물학적 지표로써의 가치에 대한 향후 연구가 의미가 있을 것으로 사료된다. Objective: The aim of this study was to determine thymosin β4 expression in epithelial ovarian cancer compared to normal ovarian tissue. Methods: Normal and pathologic ovarian tissues were obtained from healthy women (n=18), and from patients with ovarian cancer (n=27). The expression of thymosin β4 mRNA was examined by quantitative competitive polymerase chain reaction (QC PCR). Thymosin β4 mRNA expression was examined with angiopoietic factors such as vascular endothelial growth factor, angiopoietin-1 and 2. Results: The expression of thymosin β4 mRNA in epithelial ovarian cancer was higher than that in the normal ovary (p<0.05). Thymosin β4 mRNA expression was not correlated with ovarian cancer stages, pathologic types, preoperative CA125 levels, or metastasis to lymph nodes but was correlated with the expression vascular endothelial growth factor and angiopoietin-2 (p<0.05). Conclusions: Our results suggest that overexpression of thymosin β4 mRNA may be a biologic marker to differentiate epithelial ovarian cancer from normal ovary and it may play a role in angiogenesis of epithelial ovarian cancer.

Increased Expression of Thymosin β4 Is Independently Correlated with Hypoxia Inducible Factor-1α (HIF-1α) and Worse Clinical Outcome in Human Colorectal Cancer

김주헌,이승윤,박미자,이혜경,손현진,김창남,강동욱 대한병리학회 2017 Journal of Pathology and Translational Medicine Vol.51 No.1

Background: Thymosin β4 is a multi-functional hormone-like polypeptide, being involved in cell migration, angiogenesis, and tumor metastasis. This study was undertaken to clarify the clinicopathologic implications of thymosin β4 expression in human colorectal cancers (CRCs). Methods: We investigated tissue sections from 143 patients with CRC by immunohistochemistry. In addition, we evaluated the expression patterns and the clinico-pathological significance of thymosin β4 expression in association with hypoxia inducible factor-1α (HIF-1α) expression in the CRC series. Results: High expression of thymosin β4 was significantly correlated with lymphovascular invasion, invasion depth, regional lymph node metastasis, distant metastasis, and TNM stage. Patients with high expression of thymosin β4 showed poor recurrence-free survival (p = .001) and poor overall survival (p = .005) on multivariate analysis. We also found that thymosin β4 and HIF-1α were overexpressed and that thymosin β4 expression increased in parallel with HIF-1α expression in CRC. Conclusions: A high expression level of thymosin β4 indicates poor clinical outcomes and may be a useful prognostic factor in CRC. Thymosin β4 is functionally related with HIF-1α and may be a potentially valuable biomarker and possible therapeutic target for CRC.

Tissue microarray를 이용한 사이모신 베타4(Thymosin β4)와 vascular endothelial cell growth factor (VEGF)의 정상 인간 조직 발현 양상 연구

옥미선(Mee Sun Ock),차희재(Hee-Jae Cha) 한국생명과학회 2009 생명과학회지 Vol.19 No.12

사이모신 베타 4와 VEGF의 발현을 여러 인간 조직에서 tis sue microarray를 사용하여 조사하였다. 사이모신 베타 4는 간, 이자, 침샘의 관상피, 심장에서 강한 발현을 보였으며 피부, 폐, 이자, 림프절, 갑상선, 요관, 폐와 부신의 혈관 내피세포 등에서 중간 수준의 발현 양상을 보였다. VEGF의 발현 양상은 대체적으로 사이모신 베타 4와 동일하였으며 이자, 요관, 유선, 간, 식도, 신장, 폐, 부신 등의 혈관 내피세포에서 강하게 발현되었다. 이러한 결과를 통해 사이모신 베타 4는 간, 이자, 침샘의 관상피, 심장에서 중요한 역할을 담당하며 VEGF와 같은 발현 양상을 보여 혈관 신생작용에 관여함을 확인하였다. Thymosin β4, a small protein containing 43 amino acids, has multi-functional roles in cell physiology. It was first identified as a thymic maturation factor and recently has been shown to accelerate wound healing, hair growth, angiogenesis, tumor growth, and metastasis. It was also reported to play a key role in developing organs, including the nervous system and heart. Thymosin β4 induces the expression of vascular endothelial cell growth factor (VEGF), laminin-5, and other important biologically active genes. Using tissue microarray analysis, we investigated the expression patterns of thymosin β4 and VEGF in various normal human adult tissues. Thymosin β4 was highly expressed in the liver, pancreas, ductal epithelium of the salivary gland, and heart, and moderately expressed in the skin, lung, spleen, lymph node, thymus, ureter, and blood endothelial cells in both the lung and adrenal gland. The expression of VEGF generally co-localized with thymosin β4 and VEGF was highly expressed in the pancreas, ureter, mammary gland, liver, esophagus, and blood endothelial cells in both the lung and adrenal gland. These results suggest that thymosin β4 plays an important role in the function of various organs and since the expression pattern of thymosin β4 co-localized with VEGF, part of that function may be to induce or maintain angiogenesis.

Analysis of Expression Patterns of Thymosin β4 and CD133 in Normal Stomach

Mee Sun Ock(옥미선),Hee-Jae Cha(차희재) 한국생명과학회 2012 생명과학회지 Vol.22 No.10

Thymosin β4 는 대장암에서 암 줄기세포 마커인 CD133을 지닌 세포에서 지니지 않은 세포에 비해 강하게 발현된다고 보고되어 있다. 본 연구에서는 thymosin β4와 줄기세포 마커인 CD133의 상관관계를 정상 위 조직에서 관찰하였다. Thymosin β4와 CD133의 발현 양상은 tissue microarray 조직상에서 면역화학적 방법으로 관찰하였으며 thymosin β4와 CD133의 존재 위치는 면역형광 염색법 및 confocal microscope를 사용하여 조사하였다. Thymosin β4와 CD133은 동일한 양상으로 발현되었으며 모두 위의 선조직에서 강하게 발현되었다. 면역 형광 염색법으로 두가지 단백질을 동시에 염색한 결과 두 단백질이 동일한 위치에서 함께 존재하는 것으로 규명되었다. 이러한 결과는 thymosin β4와 CD133은 정상위의 선조직에서 발현되며 두 단백질의 발현 양상 및 위치가 동일하여 서로 긴밀한 상호작용을 할 가능성을 제시하고 있다. Thymosin β4 (Tβ4) has been reported to be overexpressed in CD133-positive colorectal cancer stem cells. We analyzed the relationship between Tβ4 and CD133-positive stem cells in normal stomach by examining the expression patterns of Tβ4 and CD133 in normal stomach tissues by immunohistochemical staining; co-localization of Tβ4 and CD133 was studied by immunofluorescence and confocal laser-scanning microscopy. Both Tβ4 and CD133 were expressed in stomach glands and showed similar expression patterns. Immunofluorescence staining of Tβ4 and CD133 showed that the expression of Tβ4 and CD133 was co-localized. In summary, both Tβ4 and CD133 were expressed in glands of normal stomachs and expression patterns were co-localized. These data suggest that Tβ4 expression is strongly related to CD133 expression.

An approach for identifying in silico peptides against authentic metabolites: in vitro characterization of thymosin β4 metabolites

Rahaman Khandoker Asiqur,Muresan Anca Raluca,Min Hophil,손정현,강민정,권오승 한국약제학회 2022 Journal of Pharmaceutical Investigation Vol.52 No.5

Purpose Thymosin β4 is a highly active protein that exerts multiple biological activities such as tissue repair, anti-inflammation, and cell maturation. Thymosin β4 has also been listed as a prohibited drug by the World Anti-doping Agency (WADA). Based on its biological activities, thymosin β4 has a high potential of abuse for the performance enhancement among athletes. This study aimed to investigate and characterize the metabolism of thymosin β4 in vitro system. Methods TB4 protein was metabolized in six different enzyme–buffer systems in vitro. After TB4 was metabolized with an appropriate buffer system, the resulting metabolites were detected by high resolution LC–MS/MS. The mass spectrum data of the observed metabolites were characterized in silico, and confirmed the structures based on synthesized authentic standards. Results Total 13 new metabolites, some of which were detected in more than one enzyme system, were found. This study characterized all of the detected metabolites according to their in silico m/z ions and compared our findings with synthesized standards. Finally, metabolites M1, M5, M7, M11, M12, and M13 were confirmed based on their synthesized authentic standards. Conclusion By using an approach for metabolizing a protein to detect, characterize and identify new peptide metabolites, 6 metabolites are identified among 13 expected potential metabolites. Newly detected metabolites may have the potential for biological activities after further screening compared to their parent protein.

Tissue microarray를 이용한 여러 암에서의 thymosin β4, vascular endothelial growth factor, 및 hypoxia-inducible factor-1α 발현양상 연구

Bo-Young Lee(이보영),Seung-Hyun Lee(이승현),Byung-Kwon Ahn(안병권),Mee Sun Ock(옥미선),Hee-Jae Cha(차희재) 한국생명과학회 2011 생명과학회지 Vol.21 No.3

사이모신 베타 4와 관련 단백질인 HIF-1α 및 VEGF의 발현을 여러 인간 암 조직에서 tissue microarray를 사용하여 조사하였다. 사이모신 베타 4는 골육중, 대장 선암, 식도 편평세포암, 신장 및 방광의 이행세포암, 폐암 및 간암에서 많이 발현되었으며 HIF-1α은 비강 역위성 유두종, 폐암 및 식도 편평세포암에서 강한 발현을 보였으며 대체로 발현되는 양상이나 위치가 사이모신 베타 4와 일치하는 것으로 관찰되었다. VEGF는 암 조직에서보다 암조직에 분포된 혈관내피에서 강하게 발현되는 양상을 나타내었으며 암세포에서는 사이모신 베타 4나 HIF-1α에 비해 강하게 발현되지 않았다. 위암, 간 혈관육종, 담낭 선암과 자궁 내막 선암에서 적당 수준의 VEGF 발현이 관찰되었으며 VEGF의 발현 양상 및 위치는 위암, 골육종, 지방종, 폐암, 간암, 담낭 선암, 식도 편평세포암, 대장 및 직장암, 신세포암을 포함하는 특정 암에서 사이모신 베타 4 및 HIF-1α와 일치하는 것으로 관찰되었다 Thymosin β4 (TB-4) has been reported to play a key role in tumor growth, metastasis and angiogenesis. In addition, TB-4 induced the expression of vascular endothelial growth factor (VEGF) and stabilized the hypoxia-inducible factor (HIF)-1α in melanoma cells. Although the importance of thymosin β4 in angiogenesis and metastasis has been proven, there are few studies that show the expression patterns of TB-4, VEGF and HIF-1α. This study was conducted to analyze the relationship among these proteins in various tumors. Using tissue microarray analysis, we investigated the expression patterns of TB-4, VEGF and HIF-1α in various tumors to identify the expression patterns and relationships of these proteins in certain tumors. TB-4 was highly expressed in osteosarcoma, colon adenocarcinoma, esophageal squamous cell carcinoma, kidney and urinary bladder transitional carcinoma, lung cancer, and liver cancer. HIF-1α was highly expressed in nasal cavity inverted papilloma, lung cancer, and esophageal squamous cell carcinoma. The expression patterns of TB-4 and HIF-1α were almost similar and co-localized. VEGF expression was high in the blood vessels in tumors, but usually not high in the tumors themselves. VEGF was moderately expressed in stomach cancer, liver angiosarcoma, gall bladder adenocarcinoma, and uterus endometrial adenocarcinoma. The expression patterns of VEGF shows similarities in certain tumors including stomach cancer, osteosarcoma, liposarcoma, lung cancer, liver cancer, gall bladder adenocarcinoma, esophageal squamous cell carcinoma, stomach cancer, colorectal carcinoma and renal cell carcinoma. These results suggest that the expression patterns of TB-4, HIF-1α and VEGF were co-localized and related to tumorigenesis and angiogenesis of certain tumors.

Thymosin β4 expression correlates with lymph node metastasis through hypoxia inducible factor- α induction in breast cancer

YOON, SUN YOUNG;LEE, HA REUM;PARK, YOORIM;KIM, JOO HEON;KIM, SOO YOUNG;YOON, SUK RAN;LEE, WANG JAE;CHO, BYUNG JOO;MIN, HYEYOUNG;BANG, JUNG-WOOK;PARK, HYUNJEONG;BANG, SA IK;CHO, DAEHO Sookmyung Women's University Research Institute of 2011 여성과 건강 Vol.6 No.2

Intratumoral hypoxia has been correlated with distant metastatic potential. Two hypoxia inducible factors (HIFs). HIF-1α and HIF-2α, are induced by hypoxia, and high expression of these proteins has been correlated to angiogenesis and distant metastasis. Thymosin β4 (Tβ4) is frequently highly expressed in cancer, and this overexpression correlates with malignant progression. The objective of this study was to investigate the clinical correlation of HIF-α with Tβ4 and the intracellular functional roles of Tβ4 on HIF-α activation. We examined HIF-1α, HlF-2α and Tβ4 expressions in clinical human breast carcinoma (n=70) by immunohisto-chemistry. We show that high expression of HIF-1α and HIF-2α strongly correlates with Tβ4 expression (P≤0.0001) and overexpression of Tβ4 correlates significantly with patients with lymph node metastasis (P<0.05) of human breast cancer. Additionally, we demonstrate that hypoxia up-regulates intracellular Tβ4 protein, which then affects HIF-α activity, which is the key in regulating VEGF expression. We confirmed that hypoxia-induced intracellular Tβ4 and HIF-α activities were reduced by interference of TIM expression using Tβ4 shRNA lentivirus. Vascular epidermal growth factor (VEGF)-A, a well-recognized lymphangiogenic cytokine, was also down-regulated, but VEGF-C and VEGF-D expressions were not affected. These findings suggest that the overexpression of Tβ4 is strongly associated with HIF-1α and HIF-2α expression and is also clinicopathologically involved with lymph node metastatic potential of breast cancer through the modulation of HIF-a activation and induction of VEGF-A. Ultimately, these results highlight Tβ4 as a potentially therapeutic target in malignant cancers.