索拉非尼对裸鼠高转移潜能肝癌细胞移植瘤microRNAs表达的影响及其意义
发布时间：2018-08-10 18:48
【摘要】：目的:观察索拉非尼对裸鼠移植瘤生长情况的影响并检测肿瘤组织mi RNA的表达变化,分析讨论mi RNA在索拉非尼抑制癌细胞增殖、抗血管形成及促上皮间质转化(epithelial to mesenchymal transitnio,EMT)发生等过程中的调控作用。方法:1建立裸鼠腹腔移植瘤模型:选择高转移潜能人肝癌细胞株MHCC-97H建立裸鼠皮下荷瘤模型,待荷瘤长至约1.5cm3,处死荷瘤裸鼠,取出肿瘤组织,将瘤块剪成3mm直径小块供实验用。实验裸鼠分A组(实验组)和B组(对照组),每组6只,将预先处理好的实验用瘤组织各1块埋入裸鼠腹腔建立裸鼠腹腔移植瘤模型。术后A组裸鼠每日索拉非尼30mg/kg(0.2ml)灌胃,日1次,B组给予等体积的溶剂,35天后处死所有裸鼠,取材。2对比索拉非尼治疗组及对照组肿瘤的大小,免疫组织化学染色法检测两组肿瘤组织微血管密度(microvessel density,MVD)、内源性缺氧标记物HIF-1α(Hypoxia-inducible factor-1α)、E-钙粘素(E-cadherin)及波形蛋白(Vimentin)的表达变化。3利用基因芯片技术检测实验组及对照组mi RNA表达情况,并对其结果进行生物信息学分析。Gene Ontology(GO)用来分析靶基因的分子功能。靶基因的显著性信号通路利用Kyoto Encyclopedia of Genes and Genomes(KEGG)进行分析。4通过生物信息学结果,分析并讨论mi RNA在索拉非尼抑制癌细胞增殖、抗血管形成及促使EMT发生等过程所起的调控作用。结果:1 A、B两组裸鼠腹腔成瘤率均为100%,实验期间两组裸鼠均无死亡,未见明显肉眼转移。2实验组裸鼠肿瘤体积明显小于对照组,MVD表达量明显低于对照组,上述差异均有统计学差异(P0.05)。实验组HIF-1ɑ表达量高于对照组,两组比较具备统计学差异(P0.05);与对照组相比,实验组E-cadherin的表达明显下调,而Vimentin表达上调,两组比较均有统计学差异(P0.05),提示索拉非尼促进上皮间质转化的发生。3基因芯片分析mi RNA表达谱的差异发现28个mi RNAs在实验组及对照组中差异表达,上调的mi RNAs有20个,下调的mi RNAs有8个。通过生物信息学分析结果判断:mi R-762、mi R-150-5p明显上调,参与调控索拉非尼抑制肝癌细胞增殖;mi R-142a-3p虽然上调不明显,但参与调控多个与肿瘤血管形成相关的生物学过程;上调的mi R-155-5p及下调的mi R-122-5p参与EMT的发生。结论:1索拉非尼具有抑制肿瘤生长及抗血管形成的双重作用;并致肿瘤组织缺氧,诱导肿瘤上皮间质转化的发生。2 mi R-762、mi R-150-5p参与调控索拉非尼抑制肝癌细胞增殖的过程,mi R-142a-3p参与调控抗肿瘤血管形成过程。3 mi R-155-5p及mi R-122-5p参与调控索拉非尼诱导EMT的发生过程。
[Abstract]:Aim: to observe the effect of sorafenib on the growth of xenografts in nude mice, and to detect the expression of mi RNA in tumor tissue, and to analyze and discuss the inhibitory effect of mi RNA on the proliferation of cancer cells in Solafenib. Regulation of anti-angiogenesis and (epithelial to mesenchymal translocation in epithelial stroma. Methods the tumor model was established in nude mice. The tumor model was established in nude mice with high metastatic potential human hepatoma cell line MHCC-97H. When the tumor was up to about 1.5 cm ~ 3, the nude mice were killed, the tumor tissue was removed, and the tumor was cut into a small 3mm diameter block for the experiment. Nude mice were divided into group A (experimental group) and group B (control group) with 6 rats in each group. Group A nude mice were given intragastric administration of 30mg/kg (0.2ml) once a day. After 35 days, all nude mice were killed with the same volume of solvent. The tumor size of group A was compared with that of control group. The expression of microvessel density (microvessel), endogenous hypoxia marker HIF-1 伪 (Hypoxia-inducible factor-1 伪), E-cadherin (E-cadherin) and vimentin (Vimentin) were detected by immunohistochemical staining. The results were analyzed by bioinformatics. Gene Ontology (GO) was used to analyze the molecular function of target gene. The significant signaling pathway of target gene was analyzed by Kyoto Encyclopedia of Genes and Genomes (KEGG). 4 through the bioinformatics results, we analyzed and discussed the role of mi RNA in the inhibition of cancer cell proliferation, anti-angiogenesis and the promotion of EMT by Solafenil. Results the tumorigenic rate of the nude mice in the two groups was 100. During the experiment, there was no death in the nude mice. The tumor volume of the nude mice in the experimental group was significantly smaller than that in the control group, and the expression of MVD in the experimental group was significantly lower than that in the control group. All the above differences were statistically significant (P0.05). The expression of HIF-1 in the experimental group was higher than that in the control group (P0.05). Compared with the control group, the expression of E-cadherin in the experimental group was down-regulated, while the expression of Vimentin was up-regulated. There was significant difference between the two groups (P0.05), which suggested that the difference of RNA expression profile between the two groups was found in 28 RNAs and 20 up-regulated mi RNAs in the experimental group and the control group, respectively. There are 8 down-regulated mi RNAs. The results of bioinformatics analysis showed that: mi R-762 / mi R-150-5p was significantly up-regulated, and Sorafenil was involved in regulating the biological processes related to tumor angiogenesis, although the up-regulation of sorafenil on the proliferation of hepatoma cells was not obvious. The up-regulated and down-regulated mi R-155-5p and down-regulated mi R-122-5p were involved in the pathogenesis of EMT. Conclusion: 1 Sorafenil has the dual effects of inhibiting tumor growth and anti-angiogenesis, and causing anoxia in tumor tissue. Induction of mesenchymal transformation in tumor cells. 2mi R-762mmi R-150-5p was involved in the regulation of the proliferation of liver cancer cells inhibited by Solafenil. Mi R-142a-3p was involved in the regulation of anti-tumor angiogenesis. 3 mi R-155-5p and mi R-122-5p were involved in the regulation of EMT induced by Solafenib.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R735.7
本文编号：2175874
[Abstract]:Aim: to observe the effect of sorafenib on the growth of xenografts in nude mice, and to detect the expression of mi RNA in tumor tissue, and to analyze and discuss the inhibitory effect of mi RNA on the proliferation of cancer cells in Solafenib. Regulation of anti-angiogenesis and (epithelial to mesenchymal translocation in epithelial stroma. Methods the tumor model was established in nude mice. The tumor model was established in nude mice with high metastatic potential human hepatoma cell line MHCC-97H. When the tumor was up to about 1.5 cm ~ 3, the nude mice were killed, the tumor tissue was removed, and the tumor was cut into a small 3mm diameter block for the experiment. Nude mice were divided into group A (experimental group) and group B (control group) with 6 rats in each group. Group A nude mice were given intragastric administration of 30mg/kg (0.2ml) once a day. After 35 days, all nude mice were killed with the same volume of solvent. The tumor size of group A was compared with that of control group. The expression of microvessel density (microvessel), endogenous hypoxia marker HIF-1 伪 (Hypoxia-inducible factor-1 伪), E-cadherin (E-cadherin) and vimentin (Vimentin) were detected by immunohistochemical staining. The results were analyzed by bioinformatics. Gene Ontology (GO) was used to analyze the molecular function of target gene. The significant signaling pathway of target gene was analyzed by Kyoto Encyclopedia of Genes and Genomes (KEGG). 4 through the bioinformatics results, we analyzed and discussed the role of mi RNA in the inhibition of cancer cell proliferation, anti-angiogenesis and the promotion of EMT by Solafenil. Results the tumorigenic rate of the nude mice in the two groups was 100. During the experiment, there was no death in the nude mice. The tumor volume of the nude mice in the experimental group was significantly smaller than that in the control group, and the expression of MVD in the experimental group was significantly lower than that in the control group. All the above differences were statistically significant (P0.05). The expression of HIF-1 in the experimental group was higher than that in the control group (P0.05). Compared with the control group, the expression of E-cadherin in the experimental group was down-regulated, while the expression of Vimentin was up-regulated. There was significant difference between the two groups (P0.05), which suggested that the difference of RNA expression profile between the two groups was found in 28 RNAs and 20 up-regulated mi RNAs in the experimental group and the control group, respectively. There are 8 down-regulated mi RNAs. The results of bioinformatics analysis showed that: mi R-762 / mi R-150-5p was significantly up-regulated, and Sorafenil was involved in regulating the biological processes related to tumor angiogenesis, although the up-regulation of sorafenil on the proliferation of hepatoma cells was not obvious. The up-regulated and down-regulated mi R-155-5p and down-regulated mi R-122-5p were involved in the pathogenesis of EMT. Conclusion: 1 Sorafenil has the dual effects of inhibiting tumor growth and anti-angiogenesis, and causing anoxia in tumor tissue. Induction of mesenchymal transformation in tumor cells. 2mi R-762mmi R-150-5p was involved in the regulation of the proliferation of liver cancer cells inhibited by Solafenil. Mi R-142a-3p was involved in the regulation of anti-tumor angiogenesis. 3 mi R-155-5p and mi R-122-5p were involved in the regulation of EMT induced by Solafenib.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R735.7
【参考文献】
相关期刊论文 前1条
1 宋晓;蔡振旭;;MicroRNAs的失调在高转移肝细胞癌中的作用[J];中华临床医师杂志(电子版);2013年22期
，本文编号：2175874
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