EGb761逆转NF-κB介导的胃癌细胞耐药及其机制研究

发布时间：2018-03-21 06:04

本文选题：银杏叶提取物　切入点：胃癌　出处：《广西医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:探讨银杏叶提取物(ginkgo biloba extract,EGb761)对顺铂(cis-Diaminedichloroplatinum)诱导的人胃癌细胞株SGC-7901及耐药细胞株SGC-7901/CDDP增殖和凋亡的影响及其对胃癌细胞化疗耐药的影响及其机制。方法:将实验分为SGC-7901及SGC-7901/CDDP两大组,采用EGb761、顺铂以及EGb761+顺铂联合应用处理两组细胞,采用四甲基偶氮唑蓝(methyl thiazolyl tetrazolium,MTT)法分别检测两种细胞的增殖活性,流式细胞仪Annexin V-PE/7AAD双染法检测两组细胞的凋亡,采用实时荧光定量PCR(RT-PCR)方法检测NIBP、NF-κB P65 m RNA的表达水平,采用免疫印迹法(Western blot)检测NIBP、NF-κB P65蛋白的表达水平。结果:EGb761和顺铂均对两种胃癌细胞的增殖具有显著的抑制作用,并呈剂量依赖性,但SGC-7901/CDDP细胞对顺铂的敏感性较SGC-7901细胞差。EGb761与顺铂联合应用时可明显增强SGC-7901和SGC-7901/CDDP细胞株对顺铂的敏感性并促进细胞的凋亡。实时荧光定量PCR和免疫印迹结果均显示SGC-7901/CDDP细胞株中NIBP和NF-κB p65的表达量比SGC-7901细胞株高,EGb761能明显抑制由顺铂诱导的NIBP和NF-κB p65的表达。实时荧光定量PCR结果显示,SGC-7901细胞株vs SGC-7901/CDDP细胞株,各组NIBP m RNA相对表达量(SGC-7901组:空白对照组(1.065±0.039),EGb761组(0.899±0.036),顺铂组(1.444±0.058),EGb761+顺铂组(1.107±0.040);SGC-7901/CDDP组:空白对照组(1.606±0.065),EGb761组(1.363±0.067),顺铂组(2.356±0.092),EGb761+顺铂组(1.780±0.076)),NF-κB p65 m RNA相对表达量(SGC-7901组:空白对照组(1.115±0.036),EGb761组(0.857±0.087),顺铂组(1.480±0.148),EGb761+顺铂组(1.148±0.056);SGC-7901/CDDP组:空白对照组(1.442±0.025),EGb761组(1.206±0.071),顺铂组(2.619±0.215),EGb761+顺铂组(1.634±0.072)。免疫印迹结果显示,SGC-7901细胞株vs SGC-7901/CDDP细胞株,NIBP蛋白表达量:0.324±0.021 vs 0.707±0.037,NF-κB P65蛋白表达量:0.783±0.029 vs 1.540±0.038);各组NIBP的蛋白表达(SGC-7901组:空白对照组(0.324±0.021),EGb761组(0.233±0.023),顺铂组(0.590±0.023),EGb761+顺铂组(0.328±0.022);SGC-7901/CDDP组:空白对照组(0.707±0.037),EGb761组(0.591±0.037),顺铂组(0.990±0.037),EGb761+顺铂组(0.725±0.037));NF-κB p65的蛋白表达(SGC-7901组:空白对照组(0.783±0.029),EGb761组(0.628±0.030),顺铂组(1.138±0.029),EGb761+顺铂组(0.770±0.028);SGC-7901/CDDP组:空白对照组(1.540±0.038),EGb761组(0.865±0.031),顺铂组(1.981±0.030),EGb761+顺铂组(1.508±0.016))。结论:EGb761具有显著的化疗增敏、逆转胃癌细胞耐药的作用,可增强顺铂对胃癌细胞生长的抑制作用并促进细胞凋亡。其逆转肿瘤细胞耐药的作用机制可能是通过抑制NF-κB通路的活性,减少NIBP和NF-κB p65的表达而实现。
[Abstract]:Objective: to investigate the effects of ginkgo biloba extract (EGb761) on the proliferation and apoptosis of human gastric cancer cell line SGC-7901 and drug-resistant cell line SGC-7901/CDDP induced by cisplatin cis-Diaminedichloroplatinum.Methods: the experiment was divided into two groups, SGC-7901 and SGC-7901/CDDP. EGb761, cisplatin and EGb761 cisplatin were used to treat the two groups of cells. The proliferative activity of the two cells was detected by methyl thiazolyl tetrazolium MTT, and the apoptosis of the two groups was detected by Annexin V-PE-7AAD double staining. The expression of NF- 魏 B p65 m RNA was detected by real-time quantitative PCR- PCR and the expression of NF- 魏 B p65 protein was detected by Western blot. And in a dose-dependent manner, However, the sensitivity of SGC-7901/CDDP cells to cisplatin was worse than that of SGC-7901 cells. EGb761 combined with cisplatin could significantly enhance the sensitivity of SGC-7901 and SGC-7901/CDDP cell lines to cisplatin and promote cell apoptosis. Real-time quantitative PCR and Western blotting showed that SGC-7901/CDDP. The expression level of NIBP and NF- 魏 B p65 was significantly higher than that of SGC-7901 cell line, and EGb761 could inhibit the expression of NIBP and NF- 魏 B p65 induced by cisplatin. The results of real-time fluorescence quantitative PCR showed that SGC-7901 cell line vs SGC-7901/CDDP cell line. The relative expression of NIBP m RNA in SGC-7901 group: the blank control group (1.065 卤0.039) EGb761 group (0.899 卤0.036), the cisplatin group (1.444 卤0.058) EGb761 cisplatin group (1.107 卤0.040) SGC-7901 / CDDP group: the blank control group 1.606 卤0.065Eb761 group (1.363 卤0.067), the cisplatin group 2.356 卤0.092tid EGb761 cisplatin group 1.780 卤0.0766NF- 魏 B p65m RNA relative expression and the 0.0SGC-7901 group: the blank control group 1.115 卤366-EGb761 + 0.857 卤0.057; the control group 1.480 卤0.1488b761; the control group 1.480 卤0.148b761; the control group 1.480 卤0.148b761; the control group 1.480 卤0.088b761; the control group 1.480 卤0.148b761; the control group 1.480 卤0.148b761; the control group 1.480 卤0.148b761; the control group 1.480 卤0.148b761; the control group 1.480 卤0.148b761; The expression of NIBP protein in SGC-7901 / CDDP group (1.148 卤0.056): blank control group (1.442 卤0.025), EGb761 group (1.206 卤0.071), cisplatin group (2.619 卤0.215), EGb761 cisplatin group (1.634 卤0.072). Western blot analysis showed that SGC-7901 cell line vs SGC-7901/CDDP cell line expressed NIBP protein: 0.324 卤0.021 vs 0.707 卤0.037NF- 魏 B p65 protein expression volume: + 0.783 卤0.029 vs 1.540 卤0.0381.The expression of NIBP protein in SGC-7901 group was blank. Control group (0.324 卤0.021) EGb761 group (0.233 卤0.023), cisplatin group (0.590 卤0.023) EGb761 group (0.328 卤0.022) EGb7901 / CDDP group: blank control group 0.707 卤0.037 + EGb761 group (0.591 卤0.037), cisplatin group (0.990 卤0.037) EGb761 cisplatin group (0.725 卤0.037) -NFB p65 protein expression group, blank control group (0.783 卤0.029) EGb761 group (0.628 卤0.030), cisplatin group 1.138 卤0.029 EGb761 卤0.070 卤0.070 mg 7901DP group: control group (0.783 卤0.029) EGb761 卤0.038 卤0.031 卤0.031 卤0.031 卤0.031 卤0.031 卤0.038 卤0.031 卤0.038 卤0.031. Conclusion 1. 981 卤0. 030% EGb761 Cisplatin group (1.508 卤0. 016) has significant chemosensitivity, and 1. 981 卤0. 030% EGb761 Cisplatin group has significant chemosensitivity. Reversing the drug resistance of gastric cancer cells can enhance the inhibitory effect of cisplatin on the growth of gastric cancer cells and promote cell apoptosis. The mechanism of reversing drug resistance in gastric cancer cells may be by inhibiting the activity of NF- 魏 B pathway. To reduce the expression of NIBP and NF- 魏 B p65.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.2

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