本文選題：小豆蔻明 + 炎癥��； 參考：《福建醫(yī)科大學》2014年碩士論文

【摘要】：目的 炎癥與卵巢癌的生長、侵襲、血管生成等密切相關，靶向抗炎進而抑制腫瘤細胞增殖有望成為卵巢癌治療的新途徑。小豆蔻明（Cardamonin，CAR）具有抗炎抗腫瘤及抑制哺乳動物雷帕霉素靶蛋白（mammalian target of rapamycin，mTOR）活性的作用，但mTOR是否介導了CAR的抗炎作用，從而抑制卵巢癌細胞的增殖有待證實。本研究旨在明確CAR調(diào)節(jié)炎癥抗腫瘤的作用及其相關靶標，為此類藥物的研發(fā)提供實驗依據(jù)。 方法 1細胞系 人卵巢癌細胞株SKOV3 2分組與給藥 采用脂多糖（Lipopolysaccharide，LPS）誘導SKOV3卵巢癌細胞產(chǎn)生炎癥，吡咯烷二硫代氨基甲酸酯（pyrrolidine dithiocarbamate，PDTC）阻斷核轉(zhuǎn)錄因子-κB（nuclear factor-kappaB，NF-κB）介導的炎癥途徑。設立細胞對照組、LPS組、PDTC+LPS組，分別用RAP、不同劑量CAR處理，，各藥物濃度為LPS（1μg/mL）、PDTC（100μM）、RAP（0.1μM）、CAR（100μM、30μM、10μM、3μM、1μM、0.1μM）。 3測定指標及方法 3.1顯微鏡觀察細胞形態(tài)和密度 3.2MTT法測定細胞增殖 3.3PCR法檢測炎癥因子IL-6的mRNA表達 3.5Western Blot法檢測NF-κB p65、mTOR、p-mTOR、S6K1及p-S6K1蛋白的表達 結(jié)果 1CAR可劑量依賴性抑制正常及LPS誘導的SKOV3細胞增殖； 2CAR可抑制正常及LPS誘導的SKOV3細胞IL-6mRNA和細胞核NF-κB p65蛋白表達；在正常SKOV3細胞中，低劑量CAR對IL-6mRNA表達抑制作用強于RAP和高劑量CAR（P0.01）；而在LPS預處理細胞中，RAP和高劑量CAR對IL-6的抑制作用強于低劑量CAR（P0.01）； 3LPS可激活mTOR和S6K1，顯著增加p-mTOR、p-S6K1蛋白的表達（P0.01），在正常和LPS預處理細胞中，CAR均可抑制p-mTOR、p-S6K1的蛋白表達(P0.01)，CAR對二者總蛋白的表達無顯著性影響； 4PDTC預作用可顯著抑制LPS誘導的細胞核NF-κB p65蛋白表達（P0.01），而對LPS誘導的p-mTOR、p-S6K1無明顯影響；同LPS預作用組比較，PDTC+LPS預作用后RAP和高劑量CAR對p-mTOR、p-S6K1蛋白表達的抑制作用減弱；同PDTC+LPS組比較，RAP和高劑量CAR對核NF-κB p65表達無明顯抑制作用。 結(jié)論 CAR對正常及LPS誘導的SKOV3細胞增殖均有抑制作用，機制可能與CAR降低mTOR及下游S6K1的磷酸化從而影響NF-κB與IL-6的表達有關。
[Abstract]:Purpose Inflammation is closely related to the growth, invasion and angiogenesis of ovarian cancer. Cardamonine Carr) has anti-inflammatory and anti-tumor effects and inhibits the activity of mammalian rapamycin target protein target of rapamycin. However, whether mTOR mediates the anti-inflammatory effect of CAR and thus inhibits the proliferation of ovarian cancer cells remains to be confirmed. The aim of this study was to investigate the role of CAR in the regulation of inflammation and tumor and its related targets, and to provide experimental evidence for the development of these drugs. Method 1 cell line Human ovarian cancer cell line SKOV3 2 grouping and administration Lipopolysaccharide (LPS) was used to induce inflammation in SKOV3 ovarian cancer cells. Pyrrolidine dithiocarbamate- (PDTCc) blocked the inflammatory pathway mediated by nuclear transcription factor 魏 B(nuclear factor-kappa BNF- 魏 B. The control group was treated with CAR with different doses. The concentration of each drug was LPS(1 渭 g 路mL ~ (-1) ~ 100 渭 m ~ (-1) ~ (-1) ~ (-1) 渭 M ~ (-1) ~ (-1) 渭 m ~ (-1) ~ (-1) 渭 M ~ (-1) ~ (10) 渭 m ~ (-1) ~ (-1) 渭 M ~ (-1) ~ 10 渭 M ~ (-1) ~ (10 渭 m) ~ (-1) 渭 M ~ (-1) ~ (-1) 渭 M ~ (-1). 3 determination index and method 3.1 microscopic observation of cell morphology and density Determination of cell proliferation by 3.2MTT Detection of mRNA expression of inflammatory factor IL-6 by 3.3PCR Detection of the expression of p-mTORS6K1 and p-S6K1 protein in p65 mTORA of NF- 魏 B by 3.5Western Blot Result 1CAR inhibited the proliferation of normal and LPS induced SKOV3 cells in a dose-dependent manner. 2CAR inhibited the expression of IL-6mRNA and nuclear NF- 魏 B p65 protein in normal and LPS induced SKOV3 cells, and the expression of NF- 魏 B p65 protein in normal SKOV3 cells. The inhibitory effect of low dose CAR on IL-6mRNA expression was stronger than that of RAP and high dose carotene P0.01G, but the inhibitory effect of LPS pretreated cells on IL-6 was stronger than that of low dose carotene P0.01. 3LPS activated mTOR and S6K1, significantly increased the expression of p-mTOR-p-S6K1 protein. In normal and LPS pretreated cells, car could inhibit the expression of p-mTOR-p-S6K1 protein. The expression of p0. 01 protein in nuclear NF- 魏 B p65 induced by LPS was significantly inhibited by 4PDTC preconditioning, but the expression of p-S6K1 in p-mTORN induced by LPS was not significantly affected, and the inhibitory effect of RAP and high dose CAR on the expression of p-S6K1 protein in p-mTORB p65 protein was decreased compared with that in LPS pretreated group. Compared with PDTC LPS group, rap and high dose CAR had no obvious inhibitory effect on nuclear NF- 魏 B p65 expression. Conclusion Both normal and LPS induced proliferation of SKOV3 cells were inhibited by CAR. The mechanism may be related to the decrease of phosphorylation of mTOR and downstream S6K1 by CAR, which may affect the expression of NF- 魏 B and IL-6.
【學位授予單位】：福建醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R737.31


本文編號：1776202