蜂毒肽联合吉西他滨对人胰腺癌细胞Panc-1作用的研究

发布时间：2018-02-27 02:27

本文关键词： 胰腺肿瘤蜂毒肽吉西他滨　出处：《山西医科大学》2016年硕士论文　论文类型：学位论文

【摘要】：目的:研究蜂毒肽单独应用及与吉西他滨联用对人胰腺癌细胞的作用并初步探讨蜂毒肽对人胰腺癌细胞作用的可能机制。方法:实验分为蜂毒肽单药组(16μg/m L、8μg/m L、4μg/m L、2μg/m L)、吉西他滨单药组(8μg/m L、4μg/m L、2μg/m L、1μg/m L)、两药联合组(16μg/m L蜂毒肽与8μg/m L吉西他滨联合、8μg/m L蜂毒肽与4μg/m L吉西他滨联合、4μg/m L蜂毒肽与2μg/m L吉西他滨联合、2μg/m L蜂毒肽与1μg/m L吉西他滨联合)和对照组(无药物干预,只加等量培养液)。cck-8检测法检测不同组别胰腺癌细胞Panc-1的生长增殖情况,之后使用中效原理研究两种药物间的联合作用效应;流式细胞技术(Flow Cyto Metry)检查Panc-1细胞凋亡百分比;Western-blot法检测不同药物处理情况下caspase-3、caspase-8、caspase-9、STAT3及p STAT3的表达情况。结果:蜂毒肽和吉西他滨单药即可使胰腺癌Panc-1细胞的生长受到抑制,并表现出一定的浓度依赖性,联合用药要比两药单独应用抑制作用更强,差异具有显著性(p0.05)。由中效原理计算结果显示蜂毒肽与吉西他滨联合使用表现出协同抑制效应(CI1)。吉西他滨和蜂毒肽单独使用及联合使用均可以促使Panc-1出现凋亡,和对照组的情况相比较,各用药组的凋亡百分比显著升高,而两药联合较各单药更能诱导Panc-1细胞出现凋亡,差异具有显著性(p0.05)。各药物干预组STAT3蛋白表达量与非药物干预组比较无显著差异(P0.05)。吉西他滨组和对照组中p STAT3蛋白表达量比较差异无显著性(P0.05),蜂毒肽组和联合组中p STAT3蛋白较对照组下降,以联合组中下降更为显著。和对照组相比,各用药组的Caspase-3表达量明显增加,以联合组中表达量增加最为显著。吉西他滨与对照组中Caspase-8表达量比较差异无显著性(P0.05),但蜂毒肽和联合组中Caspase-8表达显著增加。各药物干预组Caspase-9表达量和对照组Caspase-9表达相比较明显增加,以联合组能够促进Caspase-9表达最为显著。结论:我们研究表明不仅蜂毒肽单药可以抑制胰腺癌Panc-1细胞的增殖,促进其凋亡,与吉西他滨联用后两药可以协同抑制Panc-1细胞增殖并促进其凋亡,其机制与减少p STAT3的生成,激活内、外源性凋亡途径促进Caspase-3、Caspase-8、Caspase-9蛋白表达有关。
[Abstract]:Objective: to study the effect of propolis on human pancreatic cancer cells and to explore the possible mechanism of the effect of propolis peptide on human pancreatic cancer cells. Methods: the experiment was divided into 16 渭 g / m L ~ (-1) and 8 渭 g / m ~ (-1) (4 渭 g / m ~ (L)) ~ (2) 渭 g / m ~ (2 渭 g / m) of propolis peptide single drug group (n = 8 渭 g / m ~ (-1)). The combination of the two drugs: 16 渭 g / mL propolis and 8 渭 g / mL gemcitabine combined with 8 渭 g / mL and 4 渭 g / mL gemcitabine combined with 4 渭 g / L and 2 渭 g / m / L gemcitabine combined with 2 渭 g / L / L / 2 渭 g / L gemcitabine and 2 渭 g / mL / L / 2 渭 g / L / L / L and 2 渭 g / m / m / m / 1 渭 g / m / ml of gemcitabine and 8 渭 g / m / m / L gemcitabine and 4 渭 g / mL / L of gemcitabine and 2 渭 g / m / m / L gemcitabine combined with 2 渭 g / L / L of propolis and 1 渭 g / m / m / L gemcitabine. Tamabine combination) and control group (no drug intervention, The growth and proliferation of different groups of pancreatic cancer cell line Panc-1 were detected by adding the same amount of culture medium, and then the combined effect of the two drugs was studied by the medium effect principle. The expression of caspase-3, caspase-8, caspase-9, STAT3 and p STAT3 in pancreatic cancer Panc-1 cells was detected by flow Cyto test and Western-blot. Results: propolis and gemcitabine could inhibit the growth of Panc-1 cells. And showed a certain concentration dependence, the combined use of two drugs alone than the inhibitory effect is stronger, The results showed that the combined use of bee venin and gemcitabine showed synergistic inhibitory effect. The combination of gemcitabine and gemcitabine could induce the apoptosis of Panc-1, both alone and in combination with the combination of gemcitabine and gemcitabine, and the results showed that the combination of gemcitabine and gemcitabine could induce the apoptosis of Panc-1. Compared with the control group, the percentage of apoptosis in each drug group was significantly higher, and the combination of two drugs could induce apoptosis of Panc-1 cells more than the single drug. There was no significant difference in the expression of STAT3 protein between the drug intervention group and the non-drug intervention group. There was no significant difference in the expression of p STAT3 protein between the gemcitabine group and the control group, and between the propolis group and the combined group. P STAT3 protein decreased compared with the control group. Compared with the control group, the expression of Caspase-3 in the combined group was significantly increased, and the expression of Caspase-3 in the combined group was significantly higher than that in the control group. There was no significant difference in the expression of Caspase-8 between gemcitabine group and control group, but the expression of propolis peptide and Caspase-8 in combination group was significantly increased. The expression of Caspase-9 in each drug intervention group was significantly higher than that in control group. Compared with other countries, Conclusion: our study shows that not only propolis peptide can inhibit the proliferation of pancreatic cancer Panc-1 cells, but also promote its apoptosis. In combination with gemcitabine, the two drugs can synergistically inhibit the proliferation and promote the apoptosis of Panc-1 cells. The mechanism is related to the reduction of p STAT3 production, activation of endogenous and exogenous apoptotic pathway to promote the expression of Caspase-3, Caspase-8 and Caspase-9 protein.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R735.9

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