【摘要】：[目的]：本課題旨在研究高良姜對腎癌786-0和caki-1細(xì)胞的體外抗癌作用,同時(shí)對高良姜的抗癌作用機(jī)制進(jìn)行初步研究,為高良姜應(yīng)用于臨床治療腎癌提供一定的實(shí)驗(yàn)及理論依據(jù)。[方法]：采用CCK-8法分析高良姜對腎癌786-0和caki-1細(xì)胞的體外生長抑制作用。采用流式細(xì)胞儀分析高良姜誘導(dǎo)腎癌786-0和caki-1細(xì)胞凋亡作用。采用相關(guān)試劑盒檢測腎癌細(xì)胞凋亡過程中細(xì)胞內(nèi)SOD(Superoxide dismutase,超氧化物歧化酶)、T-AOC(Total antioxidation,總抗氧化能力)和MDA(Methane Dicarboxylic Aldehyde,丙二醛)的變化,探討高良姜抗腎癌的ROS(reactive oxygen species,活性氧)相關(guān)作用機(jī)制。采用劃痕實(shí)驗(yàn)和Transwell侵襲實(shí)驗(yàn)研究高良姜抑制腎癌786-0和caki-1細(xì)胞遷移和侵襲的作用。采用western blot(免疫印跡試驗(yàn))方法研究EMT (Epithelial-Mesenchymal Transition,上皮間質(zhì)轉(zhuǎn)化)相關(guān)蛋白E-cadherin、N-cadherin和vimentin變化,探討高良姜抑制腎癌細(xì)胞侵襲的作用機(jī)制。[結(jié)果]：1.CCK-8實(shí)驗(yàn)結(jié)果表明,高良姜對腎癌786-0和caki-1細(xì)胞均具有一定的增殖抑制作用,抑制作用隨高良姜濃度的增加而增強(qiáng),高良姜對于786-0和caki-1細(xì)胞增殖抑制作用呈濃度依賴性。兩種腎癌細(xì)胞增殖實(shí)驗(yàn)結(jié)果均具有顯著統(tǒng)計(jì)學(xué)差異。2.流式細(xì)胞儀檢測結(jié)果表明,高良姜促進(jìn)腎癌786-0和caki-1細(xì)胞的凋亡。與對照組相比,兩種腎癌細(xì)胞的凋亡均顯著升高,凋亡率隨高良姜濃度增加呈濃度依賴性增長。實(shí)驗(yàn)結(jié)果表明,與對照組相比,腎癌786-0和caki-1細(xì)胞內(nèi)抗氧化酶SOD活性隨高良姜濃度的增加而顯著下降。細(xì)胞內(nèi)T-AOC的活性顯著降低,MDA活性顯著增加。3.細(xì)胞劃痕實(shí)驗(yàn)和Transwell侵襲實(shí)驗(yàn)結(jié)果表明,高良姜對于腎癌786-0和caki-1細(xì)胞的遷移和侵襲具有抑制作用。劃痕試驗(yàn)表明,與對照組相比,100μM高良姜處理組可以顯著抑制786-0和caki-1細(xì)胞向劃痕區(qū)域遷移的速度。侵襲實(shí)驗(yàn)表明,l00gM高良姜處理組可以顯著抑制786-0和caki-1細(xì)胞侵襲能力。Western blot實(shí)驗(yàn)結(jié)果表明,高良姜濃度與細(xì)胞內(nèi)N-cadherin和vimentin蛋白質(zhì)表達(dá)成負(fù)相關(guān),與E-cadherin表達(dá)成正相關(guān)。隨高良姜濃度的增加,N-cadherin和vimentin逐漸下降,而E-cadherin逐漸升高。[結(jié)論]：1.高良姜具有抑制腎癌786-0和caki-1細(xì)胞增殖的作用,抑制作用有明顯的濃度依賴性,隨濃度上升抑制作用顯著增強(qiáng)。2.高良姜具有誘導(dǎo)腎癌786-0和caki-1細(xì)胞凋亡的作用。抑制作用有明顯的濃度依賴性,隨濃度上升抑制作用顯著增強(qiáng)。3.高良姜具有抑制腎癌786-0和caki-1細(xì)胞遷移和侵襲的作用。4.高良姜通過產(chǎn)生細(xì)胞內(nèi)活性氧誘導(dǎo)腎癌細(xì)胞凋亡,并通過抑制細(xì)胞間質(zhì)化抑制腎癌細(xì)胞遷移和侵襲。
[Abstract]:[objective]: the aim of this study was to study the anticancer effect of alpinia officinalis on renal cell line 786-0 and caki-1 cells in vitro, and to study the mechanism of anti-cancer effect of alpinia officinalis. To provide a certain experimental and theoretical basis for the clinical treatment of renal cell carcinoma. Methods: CCK-8 assay was used to analyze the growth inhibitory effect of alpinia officinalis on RCC 786-0 and caki-1 cells in vitro. Apoptosis of RCC 786-0 and caki-1 cells was analyzed by flow cytometry. The changes of SOD (Superoxide dismutase, superoxide dismutase (SOD (Superoxide dismutase,), total antioxidant capacity of T-AOC (Total antioxidation, and MDA (Methane Dicarboxylic Aldehyde, malondialdehyde (MDA (Methane Dicarboxylic Aldehyde,) during apoptosis of renal cell carcinoma were detected by relative kit. Objective: to investigate the mechanism of ROS (reactive oxygen species, reactive oxygen species (Ros) related to RCC. The effects of alpinia officinalis on the migration and invasion of renal carcinoma 786-0 and caki-1 cells were studied by scratch test and Transwell invasion assay. The changes of E-cadherin- N-cadherin and vimentin associated proteins of EMT (Epithelial-Mesenchymal Transition, stromal transformation) were studied by western blot (Western blotting assay, and the mechanism of Gastrodia officinalis inhibiting the invasion of renal cancer cells was investigated. [results]: the results of 1.CCK-8 assay showed that alpinia officinalis could inhibit the proliferation of 786-0 and caki-1 cells, and the inhibitory effect increased with the increase of Ginger concentration. The inhibitory effect of alpinia officinale on the proliferation of 786-0 and caki-1 cells was in a concentration dependent manner. There was significant difference between the two kinds of renal cell proliferation test results. 2. 2. The results of flow cytometry showed that alpinia officinalis could promote apoptosis of 786-0 and caki-1 cells. Compared with the control group, the apoptosis rate of the two kinds of renal cancer cells increased in a concentration-dependent manner with the increase of the concentration of alpinia officinalis. The results showed that the activities of antioxidant enzyme SOD in 786-0 and caki-1 cells decreased significantly with the increase of ginger concentration. The activity of T-AOC decreased significantly and the activity of MDA increased significantly. The results of cell scratch assay and Transwell invasion assay showed that alpinia officinalis could inhibit the migration and invasion of RCC 786-0 and caki-1 cells. The scratch test showed that the migration of 786-0 and caki-1 cells to the scratch area was significantly inhibited in the 100 渭 M alpinia officinale treatment group compared with the control group. Invasion experiment showed that l00gM could significantly inhibit the invasiveness of 786-0 and caki-1 cells. The results showed that the concentration of Ginger was negatively correlated with the expression of N-cadherin and vimentin protein in the cells. There was a positive correlation with the expression of E-cadherin. With the increase of ginger concentration, N-cadherin and vimentin decreased gradually, while E-cadherin gradually increased. [conclusion]: 1. Alpinia officinalis has the effect of inhibiting the proliferation of 786-0 and caki-1 cells in a dose-dependent manner, and the inhibitory effect is significantly increased with the increase of concentration. 2. Alpinia officinalis can induce apoptosis of 786-0 and caki-1 cells in renal carcinoma. The inhibitory effect was obviously concentration-dependent, and the inhibitory effect was significantly increased with the increase of concentration. Alpinia officinalis can inhibit the migration and invasion of 786-0 and caki-1 cells. 4. Ginger induces apoptosis of renal cell carcinoma cells by producing intracellular reactive oxygen species, and inhibits migration and invasion of renal cancer cells by inhibiting interstitial cells.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：R737.11

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