姜黄素联合紫杉醇对人口腔鳞癌细胞的增殖抑制和凋亡诱导作用

发布时间：2018-04-13 06:18

本文选题：口腔鳞癌 + 姜黄素　；参考：《山东大学》2017年硕士论文

【摘要】：姜黄素(curcumin)是从姜科、天南星科植物根茎中提取的一种天然色素,最早作为食品添加剂用于食品加工行业。近年研究发现,它具备抗癌、抗自由基、消炎、医治耐药性结核病的作用,其防癌抗癌的作用已经引起医学界的广泛兴趣,有研究表明它能参与抑制多种肿瘤细胞的生长,并且具有明显的诱导肿瘤细胞凋亡的作用。紫杉醇(paclitaxel)目前主要用于乳腺癌、卵巢癌的治疗,关于聚合微管蛋白,紫杉醇有强化作用,可以减弱肿瘤细胞的过度增殖,还可以增强肿瘤的放射敏感度,。越来越多的学者正在广泛研究其对机体多部位癌变的增殖抑制作用。本研究拟探讨姜黄素、紫杉醇以及姜黄素联合紫杉醇对离体人口腔鳞癌细胞增殖的抑制作用及细胞调亡作用,探索联合用药对人口腔鳞状细胞癌的叠加治疗效应,从而为姜黄素联合紫杉醇治疗人口腔鳞状细胞癌提供实验依据。研究目的探索姜黄素联合紫杉醇在抑制人口腔鳞癌细胞增殖和促进人口腔鳞癌细胞凋亡中的叠加效应。研究方法1.培养CAL27细胞系:复苏人口腔鳞状细胞癌CAL27细胞,进行体外胰酶的消化传代,察看培养后CAL27细胞系的状态和活性,选择处于对数生长期的CAL27细胞作为实验对象。2.观察不同浓度姜黄素或紫杉醇或二者联合用药对CAL27细胞增殖的影响。利用有差异浓度和种类的姜黄素、紫杉醇及二者联合用药作用于CAL27细胞,MTT法检测姜黄素、紫杉醇及联合用药对CAL27细胞增殖活性的影响。3.观察不同浓度姜黄素、紫杉醇和二者联合用药对CAL27细胞调亡的影响。利用有差异浓度和种类的姜黄素、紫杉醇、及二者联结用药作用于CAL27细胞,TUNEL法检测不同组CAL27细胞调亡情况的差异。4.通过Western-blot实验探讨姜黄素、紫杉醇及联合用药对凋亡蛋白分子Bcl-2、Bax和活性caspase-3的表达的影响。研究结果1.人口腔鳞癌细胞培育增殖稳定,呈贴壁生长,形态正常,肿瘤细胞较快增殖,约在培养第2天时达到对数生长期。姜黄素、紫杉醇单用于CAL27细胞时,药物浓度的提高能够强化对鳞癌细胞的增殖抑制作用,表现为浓度依赖性。2.姜黄素或紫杉醇单独作用于CAL27细胞时,细胞的增殖抑制作用随着时间增加而渐进加强,具有一定的时间依赖性。3.MTT实验表明在相同作用时间内,同浓度姜黄素联合紫杉醇相比单独用药能显著抑制CAL27细胞的增殖。4.TUNEL实验表明在相同作用时间内,同浓度姜黄素联合紫杉醇相比单独用药能显著增加CAL27细胞的凋亡数目。5.Western-blot实验表明在相同作用时间内,同浓度姜黄素联合紫杉醇相比单独用药使,Bcl-2蛋白表达明显下调、Bax2蛋白及活性Caspase-3蛋白表达明显上调。结论1.姜黄素或紫杉醇都有抑制CAL27细胞增殖的效应,联合用药具有叠加效应。2.姜黄素或紫杉醇均能诱导CAL27细胞细胞凋亡联合用药具有叠加效应。3.姜黄素或紫杉醇均能下调Bcl-2蛋白的表达,上调Bax蛋白及活性Caspase-3蛋白的表达,联合用药作用更加显著,并可以降低蛋白Bcl-2/Bax的比率。
[Abstract]:Curcumin (curcumin) from Zingiberaceae, a natural pigment extracted from Araceae plant roots, the earliest as a food additive used in food processing industry. Recent studies have found that it has anti-cancer, anti-inflammatory, anti free radical, treat drug resistant tuberculosis, its anti-cancer effect has aroused widespread interest in the medical profession. Research has shown that it can participate in the inhibition of tumor cell growth, and significantly induce apoptosis of tumor cells. Paclitaxel (paclitaxel) is mainly used for breast cancer, ovarian cancer, on the polymerization of tubulin, taxol has strengthening effect, can reduce the excessive proliferation of tumor cells, but also can enhance the tumor radiation sensitivity. More and more scholars are widely studied for many parts of the body malignant proliferation. This study intends to explore the effect of curcumin, curcumin and paclitaxel Effect of paclitaxel on apoptosis inhibition and cell proliferation of in vitro human oral squamous cell carcinoma, to explore the therapeutic effect of combined use of superposition of human oral squamous cell carcinoma, so as to provide experimental basis for curcumin combined with paclitaxel in the treatment of oral squamous cell carcinoma. The purpose of the study is to explore the effect of curcumin and paclitaxel in inhibiting and promoting effect of superposition apoptosis of human oral squamous cell carcinoma in the proliferation of human oral squamous cell carcinoma. Methods: 1. CAL27 cells: recovery of human oral squamous cell carcinoma CAL27 cells in vitro, trypsin digestion and passage, and inspect the state activity of cultured CAL27 cell lines, at the logarithmic growth phase CAL27 cells as experimental object to observe effects of different concentrations of.2. curcumin or paclitaxel or combination of the two drugs on the proliferation of CAL27 cells. The curcumin concentration and the kind of difference, and the two combined with paclitaxel Drugs acting on CAL27 cells, and detected MTT, paclitaxel and combination effects on the proliferation of CAL27 cells were observed by.3. of different concentrations of curcumin, effects of paclitaxel and combination of the two drugs on apoptosis of CAL27 cells by curcumin. There are different, and concentration of paclitaxel, and two drugs acting on CAL27 connection cells, TUNEL method was used to detect the different group of CAL27 cell death of.4. Western-blot by different experimental study of curcumin, Bcl-2 paclitaxel and combination of apoptosis proteins, Bax and activity of Caspase-3 was studied. The results of 1. human oral squamous carcinoma cell proliferation is fostering stable, adherent growth, normal morphology, rapid proliferation of tumor cells and about second days in culture reached the logarithmic growth phase. Curcumin and paclitaxel for CAL27 cells, the increase of drug concentration can strengthen the squamous cell carcinoma cell proliferation inhibition, table The concentration dependence of.2. curcumin or paclitaxel alone in CAL27 cells, cell proliferation inhibition increased with time and gradual strengthening, time-dependent.3.MTT experiments show that at the same time, compared with the same concentration of curcumin combined with paclitaxel on proliferation of.4.TUNEL experimental drug alone can significantly inhibit CAL27 cells showed that in the same time, compared with the same concentration of curcumin combined with paclitaxel alone could significantly increase the apoptosis of.5.Western-blot CAL27 cells showed that the number of experiments at the same time, the same concentration of curcumin combined with paclitaxel monotherapy compared to Bcl-2 protein expression, down-regulation of Bax2 protein and activity of Caspase-3 protein expression was significantly up-regulated. Conclusion curcumin or paclitaxel 1. alcohol has the effect of inhibiting the proliferation of CAL27 cells, in combination with the superimposed effect of curcumin or paclitaxel were.2. The expression can induce the apoptosis of CAL27 cells in combination with.3. additive effect curcumin or paclitaxel can regulate Bcl-2 protein, up-regulated Bax protein expression and activity of Caspase-3 protein, the combination effect is more significant, and can reduce the rate of protein Bcl-2/Bax.

【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.8

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