6 research outputs found
Soluble and nuclear oestrogen receptor status of advanced endometrial cancer in relation to subsequent clinical prognosis
No full textBoth soluble and nuclear oestrogen receptors have been measured in at least two separate sections from 72 endometrial. cancers and 12 normal endometria. Concentration of oestrogen receptor is shown to be, in our hands, more meaningful when expressed per unit DNA than per unit protein, whether for soluble or nuclear receptor. Endometrial cancer cells from the central part of the tumour are shown to be receptor negative more frequently than those from peripheral tumour. Thus, in large cancers, biopsies from different areas are required before a tumour can be correctly designated as receptor positive, heterogeneous or receptor negative. The intratumoral variation of receptor status may relate to poor prognosis, since patients with homogeneous receptor-positive disease survive significantly longer than those with tumours showing either heterogeneous distribution of receptor or homogeneous absence of receptor. Intratumoral variation in receptor status is found to be more common in the group of patients who are within 7 years of their menopause, than in older patient
Soluble and nuclear oestrogen receptor status of advanced endometrial cancer in relation to subsequent clinical prognosis
No full textExploring the Role of Topoisomerase II Beta in Macrophage Maturation and Pro-inflammatory Cytokine Production
No full textAlthough it is known that DNA topo IIβis required for the regulation of transcription during neural development and differentiation, it is not clear whether the enzyme is required during differentiation of human monocytes into macrophages and/or the subsequent transcription of
cytokine genes. To test this, a robust model of differentiation of monocyte-like cells into macrophage-like cells using U937 and HL-60 cells treated with phorbol 12-myristate 13-acetate (PMA) and Lipopolysaccharide (LPS) was validated. Differentiation was determined by morphological and growth characteristics and CD11b surface antigen expression as determined by flow cytometry. qRT-PCR was also used to measure mRNA transcript levels of key genes known to be up-regulated during monocyte differentiation and the secretion of pro-inflammatory cytokines produced by differentiated cells were measured using ELISA.
siRNA topo IIβknockdown did not hinder monocyte-like cells from undergoing differentiation, however experiments revealed a correlation between topo IIβknockdown and secreted TNFα, with the latter decreasing when topo IIβwas reduced. This pattern was also noted when measuring IL-1βsecretion. Similar results were seen using a Murine transgenic fibroblast cell line lacking topo IIβ, which when stimulated with LPS secreted significantly lower levels of IL-6 compared to the wild type cells. Thus topo IIβexpression is necessary for
secretion of normal levels of the cytokines, TNFα, IL-1βand IL-6 in response to LPS at certain time points. In addition in the macrophage-like state of the two cell lines, the relative levels of the βisoform (mRNA and protein) were shown to be significantly increased
compared to α, further outlining the importance of topo IIβin the differentiated state. Chromatin immuno-precipitation followed by qPCR showed however that topo IIβwas not associated at three defined proximal promoter regions of either the TNFαand IL-1βgenes, although further studies are required to rule out a direct association of topo IIβwith these and other regions of the genes.
Down regulation of topo IIβprotein using the inhibitor ICRF-193 did not hinder monocyte-like cells from undergoing differentiation either. However, contrary to the knockdown results, a 6 h pre-treatment with 1 nM ICRF-193 increased TNFαlevels in these cells, both at the
mRNA and the protein level, along with a slight increase in secreted TNFα. NF-κB, EGR2, TLR4 and TLR2 transcript levels were also increased under these conditions. Thus further studies are required to determine if these increases are due to additional cellular effects of the
drug or whether topo IIβmay play an inhibitory effect on transcription.
Thus it is clear that topo IIβplays an important role in expression of cytokines and understanding the exact nature of this requires further research that may yield potential new avenues for treatment of disease
Studies the critical period of fish oil supplementation on preventing breast cancer incidence and improving spatial learning memory performance.
No full text探討魚油補充對於減緩乳癌生成關鍵時期及對空間記憶學習之影響
懷孕飲食中若攝取較多ω6多元不飽和脂肪酸將提高胎兒日後罹患乳癌機率﹔另外胚胎與泌乳時期，正是大腦快速發育，若此時缺乏ω3多元不飽和脂肪酸，將會影響空間記憶形成。本實驗目的主要探討在胚胎泌乳期因母體攝取富含ω6但缺乏ω3多元不飽和脂肪酸之葵花油飼料下，並在不同時期給魚油補充，是否能有效降低子代罹患乳癌之風險並減緩空間記憶退化。
利用懷孕第一天的Sprague-Dawley母鼠，餵食葵花油（sunflower oil）之自製飼料下，或在胚胎泌乳期、青春期與成年期分別管餵魚油42天。出生後第55天，給予7,12-Dimethylbenzanthracene (DMBA)誘發荷爾蒙依賴型乳癌，並收集親代母鼠懷孕泌乳時期與子代雌鼠乳癌生成前後尿液，以高效能液相層析儀分析其中雌激素及其代謝產物。動物195天大時以Morris water maze檢測空間記憶學習能力，223天大時犧牲。
懷孕泌乳時期餵食葵花油之飼料，會增加子代乳癌險，而在胚胎泌乳時期給予魚油補充確實能降低子代乳癌生成，然而青春期補充魚油雖無法有效降低但卻具有延緩乳癌生成之趨勢。補充魚油者，其在空間記憶學習之表現較未補充者佳。
近年國人飲食型態改變，過高的油脂攝取量，增加罹患許多疾病的風險如乳乳癌與神經退化性疾病，故婦女於懷孕或泌乳期間，若能進行魚油的補充，將能有效降低子代日後罹患乳癌與記憶損傷的機率。The aim of this study is to evaluate the period of fish oil supplement could prevent the breast cancer incidence studied in DMBA (7,12-dimethylbenzanthracene) induced Sprague-Dawley rat. Maternal diet was given with linoleic acid-rich sunflower oil at stage of fetus to lactation without or with fish oil supplement at stage of fetus to lactation, at age of puberty, or at age of adult for 42 days. Spatial memory performance was examined by Morris water maze at age of 195 days. Animals were scarified at the age of 223 days old.
It was found that maternal dietary sunflower oil intake did increase the % of cumulative breast cancer incidence in their female offspring. While, the breast cancer incidence was reduced 50% in their female offspring via maternal fish oil supplementation during pregnancy and lactation. The fish oil supplementation during puberty did delay but not inhibit the breast cancer promotion. It is interesting to know that fish oil may enhance tumor growing, since the tumor size was larger in animals with fish oil supplementation. The spatial memory performance was improved in the animals with fish oil supplementation.
It was concluded that maternal fish oil supplementation during pregnancy and lactation may be the critical period for the breast cancer prevention on their female offspring, while, fish oil supplementation during puberty may delay the breast cancer happening.中文摘要………………………………………………………………………………1
英文摘要………………………………………………………………………………2
圖目錄 ………………………………………………………………………………Ⅰ
表目錄 ………………………………………………………………………………Ⅱ
第一章 緒論………………………………………………………………………3
第二章 文獻回顧
一、乳癌影響因子………………………………………………………………4
二、飲食與乳癌…………………………………………………………………4
三、脂肪種類對乳癌發生之影響………………………………………………5
四、多元不飽和脂肪酸代謝與乳癌生成………………………………………5
五、雌激素與乳癌………………………………………………………………6
六、雌激素代謝產物對乳癌發生之促進作用…………………………………7
七、懷孕時期母體環境與飲食對子代乳癌發生之影響………………………8
八、DMBA誘發乳癌之動物模式…………………………………………………9
1. DMBA之簡介 ……………………………………………………………9
2. DMBA誘發乳癌之條件與特性…………………………………………10
(1) 品系 …………………………………………………………………10
(2) 施打時間 ……………………………………………………………10
(3) 劑量 …………………………………………………………………10
(4) 特性 …………………………………………………………………10
九、DHA對空間記憶之影響 ……………………………………………………11
十、雌激素與空間記憶…………………………………………………………11
十一、空間記憶學習能力之行為測試…………………………………………12
第三章 實驗目的與假設………………………………………………13第四章 實驗材料與方法
一、實驗動物飼料配製 ………………………………………………………14
二、實驗動物分組 ……………………………………………………………16
三、陰道開孔之檢測……………………………………………………………18
四、DMBA專一性誘發乳癌 ……………………………………………………18
五、乳癌檢測……………………………………………………………………18
六、空間記憶測試………………………………………………………………19
1. 動物分組………………………………………………………………19
2. 實驗設備………………………………………………………………19
3. 實驗流程………………………………………………………………19
(1)水迷宮…………………………………………………………………20
(2)視覺測試………………………………………………………………20
(3)記憶保存評量…………………………………………………………20
七、樣品收集……………………………………………………………………20
1. 尿液……………………………………………………………………20
(1)收集對象………………………………………………………………20
(2)收集程序………………………………………………………………21
2. 血液……………………………………………………………………21
3. 乳腺組織與腫瘤………………………………………………………21
八、分析項目與方法……………………………………………………………22
1. 脂肪酸分析……………………………………………………………22
(1)分析項目………………………………………………………………22
(2)分析儀器………………………………………………………………22
(3)分析流程………………………………………………………………23
(4)脂肪酸萃取……………………………………………………………23
(5)脂肪酸甲基化…………………………………………………………23
(6)藥品來源………………………………………………………………23
2. 雌激素代謝產物檢測…………………………………………………24
(1)分析項目………………………………………………………………24
(2)分析儀器………………………………………………………………24
(3)分析條件………………………………………………………………24
(4)尿液純化………………………………………………………………26
(5)肌酸酐濃度測量………………………………………………………27
(6)藥品及耗材來源………………………………………………………28
第五章 結果
一、 陰道開孔時間之比較 ……………………………………………………29
二、 乳癌潛伏期與發生率 ……………………………………………………30
三、 乳癌多發性 ………………………………………………………………31
四、 空間記憶表現 ……………………………………………………………32
五、 記憶保存評量 ……………………………………………………………33
六、 總逃避時間 ………………………………………………………………34
七、 乳癌與空間學習記憶表現 ………………………………………………35
第六章 討論
一、 各母鼠對子代乳癌發生率之影響 ………………………………………36
二、 不同生命週期魚油補充對子代乳癌之影響 ……………………………37
1. 乳癌發生率……………………………………………………………37
2. 乳腺腫瘤特性之綜合比較……………………………………………38
三、 陰道開孔與乳癌之相關性 ………………………………………………38
四、 魚油補充與空間記憶表現 ………………………………………………39
五、 雌激素產物檢測 …………………………………………………………40
1. 所遇到的問題…………………………………………………………41
(1)樣品結果歧異度大……………………………………………………41
(2)留滯時間改變…………………………………………………………41
(3)牴觸代謝途徑常態……………………………………………………42
2. 可能原因………………………………………………………………43
(1)尿液樣本濃度過高……………………………………………………43
(2)尿液樣本前處理時間過長……………………………………………43
(3)加酸加熱前處理………………………………………………………44
3. 解決方法………………………………………………………………44
第七章 結論……………………………………………………………45
第八章 參考文獻 ……………………………………………………46
圖目錄
Figure 1 Estrogen metabolism ……………………………………………….7
Figure 2 Experimental Hypothesis…………………………………………. 13
Figure 3 Study design………..…………………………………………....…17
Figure 4 Latency and Cumulative mammary tumor incidence...……………30
Figure 5 Cumulative mammary tumor multiplicity .……………….………..31
Figure 6 The spatial learning memory performance by Morris water maze ...32
Figure 7 Memory retention…………………………………………..………33
Figure 8 Total escape latency of the spatial learning memory performance…34
Figure 9 The spatial learning memory performance in tumor and non-tumor
carried animals……………………………………………………...35
Figure10 The chromatography of catecholestrogen by HPLC with 8 channel electrochemical detector ………………….………..40
表目錄
Table 1 Composition of experimental diet……………………………………14
Table 2 The energy source in chow diet and experimental diet………………14
Table 3 Fatty acid composition of diet and fish oil ……….…..…...…………15
Table 4 HPLC program……....……………………………….………………25
Table 5 The potential setup in electrochemical detector in HPLC……………25
Table 6 The effect of fish oil supplementation at different stage of life on vaginal opening …………..………………………….……....………29
Table 7 The effect of dam on mammary tumor incidence of their offspring…36
Table 8 Characteristics of mammary tumor..…….…………………………...38
Table 9 The concentration of 4-methoxyestradiol in 24hrs urine collection of pregnant dam…………………..………………………………......41
Table 10 The retention time of catecholestrogen in HPLC-ECD ….…………..42
Table 11 The concentration of 4-hydroxyestradiol and 2-hydroxyestradiol concentration in 24hrs urine collection of pregnant dam.....…………4
Expression and function of osteopontin variants in HCV-related liver disease and hepatocellular carcinoma.
No full textOsteopontin (OPN) is a highly secreted multi-functional sialoprotein that is widely expressed in tissues, blood and urine. It is involved in a number of normal physiological functions, but is also significantly elevated in a number of cancers. While OPN is significantly expressed in hepatocellular carcinoma (HCC) little is known as to its role and if it is expressed in the pre-cancerous hepatitis C virus (HCV) infected liver. In this thesis we show that OPN is expressed in the liver and in HCC as three variants, the full-length protein OPN-A and two splice variants OPN-B and OPN-C. Through production of stable Huh-7 cells expressing the OPN variants, we show for the first time that all variants increase proliferation of a range of cultured hepatoma cell lines in a paracrine manner through
interactions with the cell surface OPN receptor CD44. Similarly, OPN-A (and to a lesser extent OPN-B and –C) accelerated Huh-7 derived tumor growth in a nude mouse model. We also show for the first time expression of all three OPN variants in the non-diseased liver as it was previously thought that splicing was a feature specific for tumor cells. Clinically, OPN is known to be highly expressed in HCC, however, its expression in chronic hepatitis C is not well documented. In this thesis we show that OPN mRNA expression is elevated in the HCV-infected liver with a trend towards increased expression as liver disease progresses. Consistent with an increase in mRNA, serum OPN levels were also increased in the HCV-infected liver although we could find no correlation with degree of liver disease. However, our sample size was small and this section of the thesis needs repeating with a larger HCV-infected patient cohort. Furthermore, we show that elevated OPN expression is not specific to the HCV-infected liver as OPN is also elevated in the HBV-infected and alcoholic liver suggesting that HCV does not drive OPN expression but is more likely as a result of the inflammatory process in the viral infected liver. Interestingly we also show that there is a shift of OPN expression from bile duct epithelial cells in the non-diseased liver to the hepatocyte in the HCV-infected liver which raises the question as to the role of OPN in hepatocyte transformation to facilitate the development of HCC. Our evaluation of serum OPN expression also suggests that OPN has potential as both a diagnostic and potentially prognostic biomarker for not only HCC (arising from HBV and HCV infections and alcohol abuse) but also the earlier stages of HCV-related liver disease. This work for the first time characterises the expression of all OPN variants in the liver including HCC and may be useful for identifying targeted OPN-based therapeutic approaches for HCC and other cancers. Furthermore it also suggests that monitoring OPN in chronic hepatitis C may be
useful in monitoring liver disease progression and early detection of HCC.Thesis (Ph.D.) -- University of Adelaide, School of Molecular and Biomedical Science, 201
