【摘要】：第一章大黃素衍生物A對(duì)SKOV3和SKOV3-pm4細(xì)胞遷移侵襲能力影響目的研究大黃素衍生物A對(duì)具有不同淋巴結(jié)轉(zhuǎn)移潛能的卵巢癌細(xì)胞的遷移和侵襲能力的抑制作用。方法四甲基偶氮唑藍(lán)(MTT)法檢測(cè)不同濃度大黃素衍生物A處理人卵巢漿液性乳頭狀腺癌細(xì)胞(SKO V3)和具有淋巴結(jié)定向高轉(zhuǎn)移能力的亞克隆細(xì)胞(SKOV3-pm4) 24h后細(xì)胞的增殖作用。用無細(xì)胞毒性濃度的藥物作用兩種細(xì)胞后,用細(xì)胞劃痕實(shí)驗(yàn)檢測(cè)兩種細(xì)胞的遷移能力,用Transwell小室侵襲實(shí)驗(yàn)測(cè)定細(xì)胞的侵襲能力。結(jié)果在一定濃度范圍內(nèi)大黃素衍生物A對(duì)SKOV3和SKOV3-pm4細(xì)胞的增殖均有抑制作用,24h的半數(shù)抑制濃度(IC50)分別為23.21mg/L、34.46mg/L細(xì)胞劃痕實(shí)驗(yàn)結(jié)果顯示大黃素衍生物A處理后兩種細(xì)胞的遷移能力減弱。Transwell小室侵襲實(shí)驗(yàn)結(jié)果顯示,大黃素衍生物A處理后兩種細(xì)胞的侵襲能力均受到抑制。結(jié)論大黃素衍生物A能抑制具有高淋巴結(jié)轉(zhuǎn)移潛能的卵巢癌細(xì)胞的遷移和侵襲能力。第二章細(xì)胞中Rac1/Cdc42蛋白的分布以及大黃素衍生物A對(duì)Rac1/Cdc42基因和蛋白表達(dá)的影響目的探討大黃素衍生物A抑制具有不同淋巴結(jié)轉(zhuǎn)移潛能的卵巢癌細(xì)胞的遷移和侵襲能力的可能的作用機(jī)制。方法免疫熒光法觀察兩種細(xì)胞中Racl和Cdc42蛋白的定位和分布情況。用無細(xì)胞毒性濃度的藥物作用兩種細(xì)胞后,采用實(shí)時(shí)熒光定量PCR (RT-PC R)檢測(cè)Rac1、Cdc42基因的表達(dá)情況,用蛋白免疫印跡法(Western Blot)檢測(cè)Rac1、Cdc42蛋白的表達(dá)水平。結(jié)果免疫熒光法觀察到兩種細(xì)胞中Rac1和Cdc42蛋白均廣泛分布于細(xì)胞質(zhì)中,Rac1蛋白在細(xì)胞核周圍比較密集。RT-PCR和Western Blot檢測(cè)結(jié)果顯示隨著藥物濃度2 mg/L、4mg/L、8 mg/L的梯度增加,Racl基因和蛋白的表達(dá)水平均降低,與空白對(duì)照組相比,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。而Cdc42基因和蛋白的表達(dá)變化不明顯。結(jié)論大黃素衍生物A抑制具有高淋巴結(jié)轉(zhuǎn)移潛能的卵巢癌細(xì)胞的遷移和侵襲能力的作用機(jī)制可能與下調(diào)Rac1蛋白的表達(dá)有關(guān)。
[Abstract]:Chapter 1 effects of emodin derivative A on migration and invasion of SKOV3 and SKOV3-pm4 cells objective to study the inhibitory effect of emodin derivative A on migration and invasion of ovarian cancer cells with different lymph node metastasis potential. Methods Human ovarian serous papillary adenocarcinoma cells (SKO V3) and subclone cells (SKOV3-pm4) with high lymph node metastasis were detected by (MTT) assay with different concentrations of emodin derivative A. The proliferation of posterior cells. After two kinds of cells were treated with drugs without cytotoxic concentration, the migration ability of two kinds of cells was detected by cell scratch assay, and the invasion ability of two kinds of cells was measured by Transwell chamber invasion test. Results in a certain concentration range, emodin derivative A inhibited the proliferation of SKOV3 and SKOV3-pm4 cells, and the half inhibitory concentration (IC50) at 24 h was 23.21 mg / L, respectively. The results of 34.46mg/L cell scratch test showed that the migration ability of the two kinds of cells was weakened after treatment with emodin derivative A, and the invasion ability of the two kinds of cells was inhibited by Transwell chamber invasion assay. Conclusion emodin derivative A can inhibit the migration and invasion of ovarian cancer cells with high lymph node metastasis potential. The distribution of Rac1/Cdc42 protein in cells and the effect of emodin derivative A on the expression of Rac1/Cdc42 gene and protein objective to investigate the inhibitory effect of emodin derivative A on migration of ovarian cancer cells with different lymph node metastasis potential And the possible mechanism of invasion ability. Methods the localization and distribution of Racl and Cdc42 proteins were observed by immunofluorescence. The expression of Rac1,Cdc42 gene was detected by real-time fluorescence quantitative PCR (RT-PC R), and the expression of Rac1,Cdc42 protein was detected by Western blot (Western Blot). Results the Rac1 and Cdc42 proteins were widely distributed in the cytoplasm of the two cells by immunofluorescence assay, and the Rac1 protein was dense around the nucleus. The results of RT-PCR and Western Blot detection showed that with the concentration of 2 mg/L,4mg/L,, 8 the gradient of mg/L increased and the expression of Racl gene and protein decreased, compared with the control group, the difference was statistically significant (P0.05). However, the expression of Cdc42 gene and protein did not change significantly. Conclusion the mechanism of emodin derivative A in inhibiting the migration and invasion of ovarian cancer cells with high lymph node metastasis potential may be related to the down-regulation of Rac1 protein expression.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R737.31

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