Expression of c-Met in invasive meningioma : 침윤성 뇌수막종에서 c-Met의 발현

윤수미 서울대학교 대학원 2014 국내석사

Introduction: Meningiomas are divided into three grades according to 2007 WHO classification and most meningiomas are benign (WHO grade I). However, meningiomas show high recurrence rate even after curative tumor removal and invasiveness of tumor may contribute to the high recurrence rate. Recently, c-Met, HGF, AQP1 and NKCC1 have been reported to be involved in cancer invasion. The aim of this study was to elucidate whether protein expressions of c-Met, HGF, AQP1 and NKCC1 were associated with clinicopathologic variables as well as brain and bone/scalp invasion in large scale of meningiomas. Methods: We examined immunohistochemical expression of c-Met, HGF, AQP1 and NKCC1 in 198 cases of meningiomas treated with curative tumor removal (Simpson grade I or II). Kaplan – Meier analyses were used to evaluate whether patients with meningioma had a different recurrence free survival depending on c-Met, HGF, AQP1 and NKCC1 expression status. Results: c-Met-High was observed in 18.9% (35/185) without brain invasion and 46.2% (6/13) meningiomas with brain invasion. c-Met-High significantly correlated with brain invasion. (P = 0.030) And it was also observed in 18.4% (32/174) of meningiomas without bone/scalp invasion and in 37.5% (9/24) of meningiomas with bone/scalp invasion. There was a tendency for increased c-Met-High in meningiomas with bone/scalp invasion compared with meningiomas without bone/scalp invasion, although statistical significance was not reached. (P = 0.055) HGF-High did not show statistical association with brain invasion or bone/scalp invasion. (P = 0.222, P = 0.108, respectively) On the other hand, AQP1-High showed significant inverse correlation with brain invasion. (31.9% [59/187] vs. 0% [0/11], P = 0.011) But AQP1-High showed no significant difference in bone/scalp invasion. (P = 0.812) NKCC1 did not show statistical association with brain invasion or bone/scalp invasion. (P = 0.598, P > 0.9, respectively) c-MET-high showed shorter recurrence free survival (93.467 ± 8.211 months) than c-MET-low (96.131 ± 1.911 months) however, it did not reach statistical significance. (P = 0.139) There was no association of HGF-high, AQP1-High and NKCC1-High expression with recurrence free survival. Conclusions: We demonstrated that c-Met-High was associated with brain invasion of meningiomas and that c-Met expression could be a useful predictive marker for meningioma recurrence.

Characterization of leukotriene B4 receptor 2-linked pathway leading to invasion and metastasis of human bladder and ovarian cancer cells

서지민 Graduate School, Korea University 2012 국내박사

Invasion and metastasis are predominant properties of malignant cancer cells and the main causes of treatment failure that leads to death in cancer patients. Therefore, therapeutic strategies for preventing or suppressing cancer invasion and metastasis would greatly improve survival of cancer patients. An important first step in tumor metastasis is invasion of the cancer cells through the basement membrane, which requires degradation of extracellular matrix by matrix metalloproteinases (MMPs). However, the detailed molecular mechanism for cancer invasion and metastasis remain poorly understood. In the present study, I found that BLT2, a low affinity leukotriene B4 receptor, plays an important role in invasion and metastasis, and the elevated MMP is associated with BLT2-mediated cancer progression. These observations may provide a basis for novel therapeutic targets for the development of anti-cancer treatments. 1. BLT2 is a low-affinity receptor for LTB4, a potent lipid mediator of inflammation generated from arachidonic acid (AA) via the 5-lipoxygenase (5-LO) pathway. LTB4 and its receptor may exert cancer-promoting effects that foster the progression of aggressive cancers. However, its function and mechanism in the progression of human bladder cancer have not yet been investigated. In the present study, I found that the expression of BLT2 was elevated in proportion to the tumor stage in bladder cancer specimens and metastatic bladder cancer cells. In addition, inhibition of BLT2 signaling by LY255283 or siBLT2 suppressed the invasive and metastatic potential in highly metastatic bladder cancer 253J-BV cells. Moreover, I found that up-regulation of MMP-9 may lie downstream of a BLT2-NOX1/4-ROS-NF-kB signaling cascade for the invasion and metastasis of 253J-BV cells. Together, these results suggest that BLT2 plays a pivotal, mediatory role in invasion and metastasis of aggressive bladder cancer cells, acting through a 'NOX1/4-ROS-NF-kB-MMP-9'-linked signaling pathway. 2. Lipoxygenase pathway metabolites and their cognate receptors have also been implicated in ovarian cancer progression. However, the mechanism by which BLT2 might contribute to ovarian cancer invasion and metastasis has remained unknown. In this study, I found that a BLT2, a low-affinity leukotriene B4 receptor, is highly expressed in human ovarian cancer cells, and that this receptor plays a key role in the invasiveness and metastasis through activation of signal transducer and activator of transcription-3 (STAT3) and consequent up-regulation of matrix metalloproteinase 2 (MMP-2). In addition, I observed that activation of NAD(P)H oxidase-4 (NOX4) and subsequent reactive oxygen species (ROS) generation lie downstream of BLT2, mediating the stimulation of STAT3-MMP2 cascade in this process. Taken together, these results demonstrated that a 'BLT2-NOX4-ROS-STAT3-MMP-2'-linked cascade plays a crucial role in invasiveness and metastasis of ovarian cancer cells.

Expression of Pituitary Tumor Transforming Gene-1 in placenta and its roles on trophoblast

임승묵 차의과학대학교 대학원 2014 국내석사

Placenta, which is a temporary organ during pregnancy, plays a crucial role in development and protection of fetus, and maintenance of pregnancy. Trophoblast is one of the major cells of placenta, and their invasion ability is one of important factors in early implantation and placental development. Recently, pituitary tumor transforming gene 1 (PTTG1), known as securin, was shown to be involved in proliferation as well as invasion of cancer. However, the expression of PTTG1 in placenta and their roles in invasion an proliferation of trophoblast remains unknown. Thus, the objectives are to analyze the expression of PTTG1 in placenta and demonstrate its function. Furthermore, the effect of PTTG1 on invasion activity of trophoblast and its molecular mechanism according to PTTG1 expression are validated. The expression of PTTG1 in placenta was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR), Western blot and immunofluorescence. Abilities for proliferation and invasion of trophoblast depend on PTTG1 alteration were analyzed by qRT-PCR, fluorescence-activated cell sorting analysis (FACS), invasion assay, Western blot, and zymography after small interfering RNA PTTG1 (siPTTG1) treatment. Additionally, integrin/Rho-family signaling in trophoblasts by PTTG1 alteration was analyzed. Finally, we analyzed microRNA target to PTTG1 via Web database, analyzed the expression of microRNA (miR)-186-5p in placenta and trophoblast, and demonstrated the function of PTTG1 in trophoblast by miRNA mimic and inhibitor treatment. PTTG1 expressed in placenta and trophoblast both. The expression of PTTG1 localized on mainly cytoplasm of trophoblast, especially, syncytiotrophoblast, in placenta. Decreased PTTG1 expression induce to decrease trophoblast invasion through decreased matrix metalloproteinase (MMP)-2 and MMP-9 activities (p<0.05) resulting RhoA/C expression was significantly decreased (p<0.05). Otherwise, decreased PTTG1 trigger dynamic expressions of integrin alpha (ITGA) 4, ITGA5, and integrin beta (ITGB) 1. Also, the invasion ability of trophoblast was significantly increased through increased MMP-2 and MMP-9 activities after siRNA-integrin alpha 4 (siITGA4) treatment. Furthermore, treatment of miR-186-5p mimic and inhibitor controlled invasion abilities of trophoblast by alterations of PTTG1 expression and MMP activities. Taken together, PTTG1 can control invasion ability of trophoblast via regulation of MMP activities by integrin/Rho-family signaling. In addition, PTTG1 expression and its function were regulated by miR-186-5p. These results contribute to the understanding of the roles of PTTG1 on implantation and placental development as well as useful diagnostic maker in gynecological diseases through the correlation between PTTG1 and trophoblast invasion. 태반은 임신기간 중 일시적으로 생성되는 기관으로, 태아의 발달 및 유지 그리고 보호 등에 중요한 역할을 하는 기관이다. 영양막 세포는 이러한 태반을 구성하는 주요 세포로써, 특히 영양막 세포의 침윤능은 임신 초기 착상뿐만 아니라 태반 형성에 중요한 역할을 한다. 최근, securin이라고도 알려진 뇌하수체 종양-형질전환 유전자 1 (PTTG1; Pituitary tumor transforming gene-1)은 암세포의 증식 및 침윤에도 영향을 준다고 알려져 있다. 이러한 PTTG1의 태반내 발현 및 PTTG1이 영양막 세포의 증식 및 침윤능에 미치는 영향에 대한 연구는 미비한 상태이다. 따라서, 본 연구에서는 태반 및 영양막 세포에서의 PTTG1의 발현을 확인하고, PTTG1이 영양막 세포의 증식 및 침윤능에 미치는 영향 및 그 기전을 분석하고자 하였다. PTTG1의 태반 내 발현은 qRT-PCR과 웨스턴 그리고 면역형광법을 사용하여 분석하였고, PTTG1이 영양막 세포의 증식 및 침윤능에 미치는 영향을 알아보기 위해 siRNA를 이용하여 PTTG1의 발현을 감소시킨 후 qRT-PCR, 웨스턴, 면역 형광법과, 세포주기 분석, 침윤능 분석, 자이모그래피 등을 사용하여 분석하였다, 추가적으로 siRNA Integrin alpha4 과 Rho 의 억제자인 C3 exoenzyme 은 PTTG1이 영양막 세포의 침윤능에 미치는 영향에 대한 메커니즘 분석을 위해 사용하였고, PTTG1을 타겟 한다고 예상되는 miRNA cadidate를 찾아 miR-186-5p를 이용하여 다시 한번 PTTG1의 기능을 검증하였다. 본 연구 결과, PTTG1은 태반과 영양막 세포 모두에서 발현되었고, 태반에서 PTTG1의 발현은 대부분 영양막세포의 세포질에서 발현 됨을 확인할 수 있었다. 또한, 영양막 세포내 감소된 PTTG1은 MMP-2와 MMP-9 의 활성도 그리고 Rho A/C 를 감소시키는 것을 통해 영양막 세포의 침윤능을 현저히 감소시켰다. 그밖에도, PTTG1의 감소는 Integrin alpha4와 alpha5, beta1의 발현을 변화시켰고, 그 중에서도 integrin alpha4는 PTTG1이 감소될 때 현저히 증가되고, siRNA를 통해 integrin alpha4를 감소시켰을 때, MMP-2와 MMP-9의 활성을 증가시키는 것을 통해 영양막 세포의 침윤능을 증가시켰다. 추가적으로, PTTG1을 조절하는 microRNA candidate중 miR-186-5p의 발현은 PTTG1의 발현과 MMP의 활성 뿐만 아니라 영양막 세포의 침윤능 또한 조절하였다. 결론적으로 PTTG1은 Integrin/Rho 신호전달 체계를 이용해 MMP의 활성을 조절하고 이를 통해 영양막 세포의 침윤능을 조절할 수도 있는 중요한 인자라는 것을 확인하였고 추가로 miR-186-5p가 PTTG1의 발현 및 기능을 조절하는 조절자라는 것을 확인하였다. 이러한 결과는 PTTG1과 영양막 세포의 침윤능의 상관관계를 통해 착상 및 태반발달을 이해하는 기초자료로써 활용될수 있을 뿐만 아니라 다양한 산과질환의 진단 마커로써의 활용가능성을 보여주는 자료라고 사료된다.

Role of BLT2-linked cascade in the invasion and metastasis of cancer cells

김은영 School of Life Sciences and Biotechnology, Korea U 2010 국내박사

Invasion and metastasis is the critical steps in cancer progression that lead to death from this disease. Although recent reports have emphasized the importance of secreted proteases, cellular adhesion molecules, and the presence of mitogenic and angiogenic growth factors at the tumor site the details of the molecular basis for cancer invasion and metastasis remain poorly defined. In the present study, I found that BLT2, a low-affinity leukotriene B4 receptor, plays an important role in cancer progression, such as invasion and metastasis, and the elevated MMP-9 is associated with BLT2-mediated cancer progression. These observations may provide a basis for novel therapeutic targets for the development of anti-cancer treatments. 1. Ras signaling pathways are well-recognized for their involvement in cancer cell proliferation; however, considerably less is known regarding their contribution to invasion and metastasis. In the present study, I investigated whether BLT2, a low-affinity leukotriene B4 receptor, is involved in H-RasV12-evoked invasion and metastasis. I showed that a novel BLT2, a low-affinity leukotriene B4 receptor,-linked signaling cascade involving generation of reactive oxygen species (ROS) via Nox1, NF-κB stimulation, and subsequent up-regulation of matrix metalloproteinase-9 (MMP-9) is a potential mechanism by which Ras promotes invasion and metastasis. Moreover, inhibition of BLT2 signaling markedly suppressed Ras-evoked metastasis and reduced the associated mortality in mice. Consistent with the proposed role of BLT2 as a key downstream mediator of Ras signaling to metastasis, BLT2 expression alone resulted in the formation of numerous metastatic lung nodules and the nodules formation was significantly attenuated by inhibition of MMP-9, a downstream component of BLT2. Together, our results reveal the previously unsuspected function of BLT2-linked cascade in driving oncogenic Ras-induced metastasis and would provide a valuable insight into invasion and metastasis. 2. Leukotriene B4 and its receptor may exert cancer-promoting effects that foster the progression of aggressive cancers. In the present study, I found that the leukotriene B4 receptor BLT2 is appropriately overexpressed in advanced human bladder cancers and has high prognostic significance. Blockade of BLT2 suppressed the invasiveness of highly aggressive 253J-BV bladder cancer cells, whereas stimulation of BLT2 led to NAD(P)H oxidase-1 and -4 (Nox1/4)-induced generation of reactive oxygen species (ROS) and NF-κB-mediated upregulation of matrix metalloproteinase-9 (MMP-9). Metastasis of 253J-BV cells in mice was also dramatically suppressed by inhibition of BLT2 or its signaling. Moreover, I found that immunohistochemical examination of 85 cases of transitional cell carcinoma (TCC) revealed that BLT2 and MMP 9 levels are significantly correlated in high-grade, deeply invasive carcinomas (>pT2), in consistent with the idea that MMP-9 is a key downstream mediator of BLT2. These findings suggest a ‘BLT2-Nox1/4-ROS-NF-kB-MMP-9’ cascade plays a critical role in bladder cancer invasion and metastasis.

Role of Ca2+ signaling in collective migration and invasion of ameloblastoma

박성호 Graduate School, Yonsei University 2020 국내박사

Collective invasion 에 대한 연구는 암의 보존적 치료와 장기 예후에 매우 중요하다. 특히, 악성 구강암 및 법랑모세포종에서는 유방암 또는 폐암 등의 다른 암과 비교하여 칼슘과 collective invasion 사이의 명확한 기전이 알려져 있지 않아 추가 연구가 필요하다. 따라서, 본 연구의 목표는 법랑모세포종에서 collective invasion 과정에 관여하는 Ca2+ 신호의 역할을 연구하는 것이다. 본 연구에서는 인간에서 유래된 Immortalized ameloblastoma cell line (AM-1)을 사용 하였다. CaCl2, Calcium pump inhibitor (SKF96365) 및 Calcium sensing receptor inhibitor (NPS2143)을 AM-1 배양 배지에 첨가하여 세포 내 Ca2+ 농도를 제어 하였다. Ca2+ 신호의 제어 하에 AM-1세포의 세포 형태, 이동 및 침습 패턴을 분석하기 위해 Time-lapse imaging, Tumor spheroid invasion collagen assay, Immunofluorescence staining 을 사용 하였다. AM-1 세포는 Ca2+ 농도와 관련된 각기 다른 배양 조건에서 상이한 세포 형태 및 이동 패턴을 보여 주었다. 그리고 AM-1 세포는 Ca2+ 의 농도가 증가함에 따라 서로 더 밀접하게 부착되고 collective migration의 패턴을 보였다. 그러나 Ca2+ 농도를 제어 했을 때, NPS2143이 아닌 SKF96365에 의해 Ca2+ 유도 cell-cell adhesion 이 억제되었다. 또한, SKF96365에 의해 AM-1 세포의 Ca2+ 유도 collective invasion도 억제되었다. 최종적으로, Calcium pump inhibitor는 AM-1 세포의 cell-cell adhesion과 collective invasion 을 억제하는 역할을 하였다. 따라서, 세포막의 칼슘 펌프는 법랑모세포종의 collective invasion 을 억제하기 위한 잠재적인 치료 표적이 될 것으로 결론 지을 수 있다. Research on collective invasion is considered to be very important for conservative treatment and long-term prognosis of cancer. Particularly in malignant oral cancer and ameloblastoma, compared to breast cancer or lung cancer, the clear mechanism between calcium and collective invasion is unknown, and further studies are needed. Therefore, my objective is to investigate the role of Ca2+ signaling in ameloblastoma during the collective invasion process. Human derived ameloblastoma cell line (AM-1) was used in this study. CaCl2, SKF96365 and NPS2143 were added into the AM-1 culture medium to control the intracellular Ca2+ concentration. Time-lapse imaging, tumor spheroid invasion collagen assay and immunofluorescence staining were used to observe the cell morphology, migration and invasion pattern of AM-1 under the control of Ca2+ signaling. AM-1 cell showed a disparate cell morphology and migration pattern in different culture condition which is associated with Ca2+ concentration. Furthermore, AM-1 cell attached to each other more tightly and migrated collectively in Ca2+ dose-dependant manner. However, Ca2+ induced cell-cell adhesion could be suppressed by SKF96365 not NPS2143. Moreover, Ca2+ induced collective migration of AM-1 cell also can be suppressed by SKF96365. Finally, SKF96365 was sufficient to inhibit the cell-cell adhesion and collective invasion of AM-1 cell. Therefore, it can be concluded that calcium pump is expected to be a potential candidate target for inhibiting the collective invasion of ameloblastoma.

Cofilin phosphorylation contributes to invasive development of bladder cancer

강효문 건국대학교 대학원 2010 국내박사

Tumor invasion has detriment effects during the stages of cancer development. The migration of tumor cells is a prerequisite for tumor cell invasion development. Actin cytoskeletal reorganization is crucial for tumor cell migration and actin depolymerizing factor cofilin is a key regulator of actin filament turnover and cytoskeleton reorganization. The role of cofilin in cell motility has been demonstrated in some report, but remained poorly documented in bladder cancer. In this study, I examined the relationship between cancer invasion and expression of phosphor-cofilin and cofilin in tissue and cells. Proteomics technology has demonstrated its powerful potential in the filed of the pathogenesis and pathophysiology of various diseases. Tissue analysis, using 2-DE and MALDI-TOF/TOF was performed with normal bladder and cancer tissues from non-invasion and invasion bladder cancer tissue to determine the pathogenic mechanisms and new protein targets of bladder cancer. The expression of 13 proteins was altered in non-invasion and invasion compared with normal bladder tissue. Among these proteins, cofilin involved in cytoskeletal reorganization brought to our attention, I found that the expression of cofilin was increased and its isoelectric points descend in bladder tumor tissue compared with normal bladder tissue. These alterations reveal that phosphor-cofilin may be involved in invasion progress of cancer cell. In order to confirm this hypothesis, using western blot and immunohistochemical method, I examined the expression of phosphor-cofilin and cofilin in tissues mentioned above. The expressions of phospho-cofilin and cofilin were increased in non-invasion and invasion bladder cancer compared with normal bladder cancer tissue. And the expression of phospho-cofilin was increased more in invasion bladder tumor than in non-invasion bladder tumor as we expected. Analysis in cells was performed with bladder cancer cell line T24 to examine the cell invasion phenomenon-related migration and the expression of phosphorylated cofilin and cofilin in EGF-treated T24 cells. I found that T24 cell migration was induced by EGF treatment. Expression of phosphor-cofilin was transient increased by EGF treatment. But cofilin-siRNA transfection into cells decreased the expression of EGF-induced phosphorylated cofilin in the cells and reversed EGF-induced cell migration. These results suggest that protein of phosphor-cofilin may play an important role in cancer invasion development. Moreover, phosphor-cofilin may be useful for a novel biomarker in transitional cell carcinoma and provide a new strategy for the therapy of bladder cancer. 종양침윤은 암발생 과정에서 중요한 기능을 발휘한다. 종양세포의 이동은 종양세포의 침윤발생을 위한 필수요소이다. Actin에 의한 세포골격 재조직화는 종양세포의 이동을 위해 중요하다. 세포운동성에 있어서 cofilin의 역할은 일부 보고에서 보여져 왔으나 방광암에 있어는 거의 보고된 바 없다. 나는 여기서 조직 및 세포에서의 cofilin 및 인산화된 cofilin의 발현정도와 암침윤사이의 관계를 알아보기 위하여 연구를 수행했다. 단백질체학 기술은 많은 질병의 병태생리학의 분야에서 강력한 가능성을 나타내어 왔다. 따라서, 방광암의 병인 기전 및 새로운 단백질 표적들을 밝히기 위하여 2-DE 및 MALDI-TOF/TOF을 이용하여 정상, 비침윤성 및 침윤성 방광조직의 단백질 발현 프로파일을 분석하였다. 그 결과, 정상방광조직에 비해 비침윤성 및 침윤성 조직에서 13개 단백질의 발현변화가 관찰되었다. 이들 변화된 단백질 중에 세포골격의 재조직와 관련된 cofilin은 정상조직에 비해 방광종양 조직에서 그 발현이 증가 했을 뿐만아니라 그것의 등전점이 감소했다. 이들 변화는 cofilin인산화가 방광종양의 침윤과정에 관련될 가능성을 제시한다. 우리의 가설을 확인하기 위해 western blot법 및 면역조직화학 염색법을 이용하여 cofilin 및 인산화된 cofilin의 발현이 정상과 침윤성 및 비침윤성 암조직에서 검사되었다. 인산화된 cofilin과 cofilin은 정상 방광암조직에 비해 비침윤성 및 침윤성 방광암조직에서 증가 되었다. 그리고 인산화된 cofilin은 비침윤성 조직에서보다 침윤성 조직에서 더 증가 하였으며, 이 결과는 우리의 예상과 일치했다. EGF로 자극한 T24 방광암세포의 분석에서 인산화된 cofilin 및 cofilin의 발현정도 및 세포의 침윤과 관련된 이동현상을 관찰하였다. T24 세포에 EGF의 처리는 세포이동을 발생시켰으며, 인산화된 cofilin의 발현을 일시적으로 증가시켰다. 그러나 cofilin siRNA 처리한 세포에서는 인산화된 cofilin의 발현이 감소되었다. 이상의 결과들로부터, 인산화된 cofilin이 암의 침윤발생에 중요한 역할을 할지도 모른다는 것이 시사되었다. 따라서 인산화된 cofilin은 방광암 치료를 위한 새로운 제제의 개발을 위한 후보물질로서 뿐만아니라, 전이성 세포암종에 대한 새로운 biomarker로서 유용한 물질일 것으로 사료된다.

췌장선암 정맥침범 마커로서의 TIMP1

성유나 울산대학교 대학원 2023 국내석사

Background Pancreatic ductal adenocarcinoma (PDAC) is the 8th most common cancer in Korea and has a low 5-year overall survival rate (YSR) of about 12.2%. Although venous invasion is known to be the cause of poor prognosis, the precise mechanism is still unknown. In this study, we investigated biomarkers involved in venous invasion and their mechanisms using gene expression arrays and functional validation. Materials and methods Eight surgically resected formalin-fixed, paraffin-embedded (FFPE) PDAC tissues were collected. Meticulous manual microdissections for gene expression arrays were performed on the following three groups. 1) portal vein/ superior mesenteric vein with cancer cell invasion (VI); 2) pancreatic cancer without portal vein/ superior mesenteric vein invasion (CA); and 3) normal portal vein/ superior mesenteric vein tissue (NV). The candidate gene expressions were validated at protein level in 220 cases using 2D images with immunohistochemistry (IHC) and 3D images with tissue clearing and multiple immunofluorescence labeling. To identify the role of potential biomarker in venous invasion, invasion assay and western blot analysis were performed using human endothelial (EA.hy926), cancer-associated fibroblast (CAF), and pancreatic cancer (Panc1) cell lines. Results Four genes, includingTIMP1, CXCR4, OLFML2B, and CYP1B1, were specifically expressed in VI group. TIMP1 (p = 0.026) and CXCR4 protein (p < 0.001) expression in VI set were significantly higher than in CA set. Specific TIMP1 expression in venous invasive areas was found by 3D imaging. The patients with strong TIMP1-expression more commonly had lymphovascular invasion (p < 0.001) and low 5-YSR (p = 0.027) than those with weak TIMP1-expression. TIMP1 inhibition by siRNA reduced cancer cell invasion ability in the presence of CAF. TIMP1 was increased in pancreatic cancer cells along with PI3Kp110 and phospho-AKT in co-culture conditions mimicking venous invasion. Conclusions TIMP1 was a potential biomarker of venous invasion in PDAC and the TIMP1/PI3K/Akt signaling pathway may be involved. This could provide basic information for development of inhibitors to prevent venous invasion in patients with PDAC.

Historical and Genetic Approaches to Understanding Invasion Success of the European Starling (Sturnus Vulgaris) : ENFOQUES HISTORICO Y GENETICO PARA COMPRENDER EL EXITO DE LA INVASION DEL ESTORNINO PINTO (STURNUS VULGARIS)

Hofmeister, Natalie R ProQuest Dissertations & Theses Cornell University 2022 해외박사(DDOD)

How invasive species establish and spread is a central question in biology, but also one that impacts public perceptions of and engagement with those species. I use the invasive European starling (Sturnus vulgaris) as a case study to identify factors supporting its establishment and expansion in the United States, and to consider its evolutionary history in its native range. In my first chapter, I integrate recent work in the evolutionary genetics of invasive European starling populations with ecological studies over its residence in each region, identifying areas for future research. In my second chapter, I use genomic methods to reconstruct demographic history and test for natural selection in the North American invasion, and I find that starlings in North America show evidence for local adaptation despite a genetic bottleneck upon invasion. In my third chapter, I compare patterns of genetic variation between the concurrent North American and Australian invasions: starling populations show remarkably high differentiation from each other on a short evolutionary timescale, and this differentiation is consistent with selection in at least a few regions of the genome. In my fourth chapter, I consider starling invasions from the perspective of science and technology studies (STS), tracing where human interference and potential biases shape both the practice of invasion science. I consider how my own genomic analyses depend on assumptions in both population genetic methods and invasion theory. Overall, this dissertation attempts to reconstruct the evolutionary history of starling invasions, with a focus on the invasion in North America.

Mitochondrial DNA-depleted HeLa 세포에서의 Salmonella invasion과 proliferation에 대한 연구 : Studies on the Salmonella Invasion and Proliferation in Mitochondrial DNA-depleted HeLa Cells

천성균 건국대학교 2004 국내석사

한글초록:Salmonella의 병인 기전은 주로 병원성 물질과 숙주세포와 Salmonella의 관계를 위주로 연구 되어졌고, 아직까지 Salmonella invasion과 proliferation에 관해서는 알려진 것이 거의 없다. 또한 숙주세포 mitochondria와 Salmonella invasion 그리고 proliferation의 관계에 대해 연구 되어진 것이 없으므로 mitochondria가 Salmonella invasion과 proliferation에 있어 하나의 요인으로 작용할 것이라는 가정하에 본 연구를 수행하였다. Mitochondria의 기능을 저해는 ethidium bromide (EtBr)를 HeLa 세포에 처리하는 방법을 사용하였고 4주 동안 EtBr을 처리하여 mitochondrial DNA-depleted HeLa (ρ0) 세포를 만들었다. 이 세포를 이용하여 Salmonella invasion assay, proliferation assay 그리고 immunostaning-fluorescence microscopy를 실시하였다. Immunostaning을 실시하여 normal HeLa에 비해 ρ0 세포는 mitochondria가 염색되지 않았다. Normal HeLa에 비해 ρ0 세포는 40% 정도 Salmonella가 invasion 되었고, Salmonella proliferation은 48시간 후 약 두 배 적게 proliferation 되었다 영문초록: Salmonella pathogenesis studies have been focused on virulence factors and their roles in host-Salmonella interactions, because host factors which could determine the susceptibility to Salmonella invasion are rarely known. We hypothesized that mitochondria function might be critical roles in invasion mechanism. In order to identify this objectives, we established mitochondrial DNA (mtDNA)-depleted HeLa cells (rho0, ρ0) by treating ethidium bromide (3,8-diamino-5-ethyl-6-phenylphenanthridinium bromide, EtBr). With this deleted cells performed invasion assay, proliferation assay and immunostaining-fluorescence microscopy with the normal HeLa cells and the ρ0 cells. The invasion rate of Salmonella into ρ0 cells was approximately 40% of that of Salmonella into normal HeLa cells. Salmonella proliferation rate was higher in the normal HeLa cells than in ρ0 cells. Our results suggest that mitochondria is one of an important factor to protect their invasion and proliferation of Salmonella in host cells

Anti-cancer effect of oleanolic acid on oral squamous cell carcinoma

양혜지 Graduate School, Yonsei University 2023 국내석사

Oral cancer is a common malignancy in the worldwide and the 5-year survival rate of oral cancer patients has remained at about 50% in the last few years. Oleanolic acid is a pentacyclic triterpenoid from medicinal plants, including Olea europaea, Hyptis capitata, and Rosa woodsii. Although anti-cancer activity of oleanolic acid has been reported in several cancers, its inhibitory activity on OSCC cell invasion still remains unclear. In this study, I investigated the inhibitory effect of oleanolic acid on OSCC cell invasion and its underlying molecular mechanisms. Treatment with oleanolic acid decreased cell viability and proliferation in Ca9-22 and YD-10B OSCC cells stimulated with or without plasminogen activator inhibitor (PAI)-1. Treatment with oleanolic acid induced apoptotic and necrotic cell death but oleanolic acidinduced cell death was blocked by PAI-1-stimulation in these OSCC cells. PAI-1-stimulation promoted cell migration and invasion but oleanolic acid treatment significantly suppressed PAI-1-stimulated cell migration and invasion in Ca9-22 and YD-10B OSCC cells. Oleanolic acid decreased the expression of an epithelial cell marker zonula occludens-1, and mesenchymal cell markers α-smooth muscle actin and vimentin, in OSCC cells stimulated with or without PAI-1. In addition, PAI-1-stimulation promoted the secretion of C-X-C motif chemokine ligand (CXCL) 10 but the increased CXCL10 secretion by PAI-1-stimulation was inhibited by treatment with oleanolic acid in OSCC cells. Treatment with neutralizing antibody against CXCL10 or its receptor CXCR3 remarkably inhibited PAI-1-stimulated invasion in OSCC cells. CXCL10 promoted OSCC cell invasion but CXCL10-stimulated OSCC cell invasion was suppressed by treatment with oleanolic acid. Treatment with JSH-23, a selective inhibitor of NF-κB and MK-2206, a selective inhibitor of AKT, blocked cell invasion in OSCC cells with or without PAI-1. In conclusion, oleanolic acid suppresses cell viability, proliferation, migration, and invasion in OSCC cells stimulated with or without PAI-1. PAI-1 promotes OSCC cell invasion by increasing CXCL10 production and activating AKT/NF-κB signaling and oleanolic acid inhibits cell invasion by blocking PAI-1-stimulated CXCL10 production and CXCL10-stimulated OSCC cell invasion. Therefore, oleanolic acid may be potential anti-cancer agent against OSCC cell invasion.