【作者】 欧阳华强；

【导师】 刘鲁明； 于尔辛； 陈震；

【作者基本信息】 复旦大学 ， 中西医结合临床， 2009， 博士

【摘要】 目的了解清胰化积方对人胰腺癌CFPAC-1裸鼠移植瘤的抑瘤作用,从细胞因子、细胞周期、癌相关基因表达层面探索其抑瘤机理。方法1.清胰化积方对人胰腺癌CFPAC-1裸鼠移植瘤的抑瘤作用研究采用荷CFPAC-1人胰腺癌裸小鼠模型,随机分组后,以不同剂量的清胰化积方及化疗药物进行干预,观察药物的体内抑瘤作用。2.清胰化积中药对人胰腺癌CFPAC-1裸鼠移植瘤的抑瘤机理探讨2.1采用ELISA法检测裸鼠血清细胞因子VEGF、TNF-α、sVCAM-1、TGF-β1、IL-6和IL-8的表达;2.2用流式细胞术检测清胰化积方对CFPAC-1裸鼠移植瘤细胞周期的影响;2.3用普通RT-PCR及qPCR法检测EphB2的表达,并针对目标基因EphB2构建慢病毒干扰载体,进行RNA干扰等深入研究。结果1.清胰化积方对人胰腺癌CFPAC-1裸鼠移植瘤的抑瘤作用研究清胰化积方对人胰腺癌CFPAC-1细胞株体内生长有一定的抑制作用,中药组瘤重低于对照组（p＜0.05）,清胰化积方各剂量组的抑瘤率约32～34%。2.清胰化积中药对人胰腺癌CFPAC-1裸鼠移植瘤的抑瘤机理探讨2.1清胰化积方对人胰腺癌CFPAC-1裸鼠血清VEGF、TNF-α、sVCAM-1、TGF-β1、IL-6和IL-8表达水平的影响:清胰化积方治疗后相应组裸鼠血清TNF-α、IL-6、IL-8水平显著低于对照组（p＜0.05）,且与瘤重呈正相关。各组裸鼠血清VEGF、TGF-β1、和sVCAM-1表达水平之间无显著性差异（P＞0.05）。2.2清胰化积方对人胰腺癌CFPAC-1裸鼠移植瘤细胞周期的影响:清胰化积方治疗后CFPAC-1细胞出现S期阻滞,S期细胞比例明显高于对照组,G₂/M期细胞比例低于对照组（P＜0.01）。各组间的凋亡率则无显著性差异（P＞0.05）。2.3清胰化积方对人胰腺癌CFPAC-1裸鼠移植瘤EphB2表达的影响:清胰化积方组EphB2表达水平与对照组相比较有两倍以上的上调。针对CFPAC-1细胞系进行EphB2 RNA干扰后,流式细胞分析提示EphB2下调可以抑制CFPAC-1细胞的凋亡。结论1.清胰化积方对人胰腺癌CFPAC-1体内生长有明显的抑制作用,主要表现为诱导胰腺癌细胞发生坏死。2.清胰化积方抑瘤作用的机理,可能与减少人胰腺癌CFPAC-1荷瘤裸鼠血清细胞因子含量、诱导肿瘤细胞周期的S期阻滞及上调EphB2基因表达有关。更多还原

【Abstract】 ObjectivesThe aim of this study was to investigate the inhibitory effects of QYHJ decoction on human pancreatic cancer cell line CFPAC-1 in nude mice,and its influence on extracellular factors,cell cycle,cancer related genes and signal transduction.Materials and Methods1.Reaseach on Inhibitory effects of QYHJ decoction on experimental pancreatic cancer cell line CFPAC-1 in nude miceHuman pancreatic cancer cell line CFPAC-1 burdened nude mice were randomized to receive QYHJ decoction at different doses and oral chemotherapeutics.Inhibitory rates of tumor growth in vivo were calculated.2.Research on mechanism of inhibitory effects of QYHJ decoction on experimental pancreatic cancer cell line CFPAC-1 in nude mice2.1 ELISA was used to detect serum extracellular factors such as VEGF、TNF-α、sVCAM-1、TGF-β1、IL-6 and IL-8 in nude mice.2.2 Flow cytometry technique was used to analyze transplanted tumors of nude mice. RT-PCR and quantitative PCR were used to amplify the gene of EphB2. Lentivirus interference vector was constructed to knockdown the targeted gene EphB2 for further study. Results1.Inhibitory effects of QYHJ decoction on experimental pancreatic cancer cell line CFPAC-1 in nude miceQYHJ decoction could inhibit the growth of human pancreatic cancer cell line CFPAC-1 in vivo.Tumor weight in the QYHJ groups was significantly lower than that in control group(P＜0.05).Inhibiting rate of tumor growth varied from 33 to 38%in different dose of QYHJ groups.2.mechanism of inhibitory effects of QYHJ decoction on experimental pancreatic cancer cell line CFPAC-1 in nude mice2.1 The expression of serum extracellular factors such as VEGF、TNF-α、sVCAM-1、TGF-β1、IL-6 and IL-8 in nude mice:The serum level of TNF-α, IL-6 and IL-8 in the QYHJ-treated group was significantly lower than that in the control group(P＜0.05),and these above factors had a negative correlation with the inhibitory rate.There were no significant difference among each groups for the serum level of VEGF、TGF-β1 and sVCAM-1(P＞0.05).2.2 Results of cytometry analyses in each group:S-phase arrest was detected in the tumors of QYHJ-treated groups:The proportion of cells in S phase was higher, and which in G2/M phase was lower accordingly,when compared with the control group(P＜0.05).2.3 The expression of EphB2 in transplanted tumor of each group:The expression level of EphB2 in QYHJ-treated group was 2-fold or more elevated than that in the control group.Cytometry analyses revealed that downregulation of EphB2 may promote apoptosis in the CFPAC-1 cell line. Conclusions1.QYHJ decoction may remarkablely inhibit the growth of human pancreatic cancer cell line CFPAC-1 in vivo,the pattern of manifestation is to induce the pancreatic cancer cells to be necrosis.2.The mechanism of inhibitory effects of QYHJ decoction may correlate with downregulation of the serum level of extracellular factors,S-phase arrest in the cell cycle induced by QYHJ decoction,and upregulation of EphB2.更多还原