【摘要】：目的:觀察索拉非尼對裸鼠移植瘤生長情況的影響并檢測腫瘤組織mi RNA的表達(dá)變化,分析討論mi RNA在索拉非尼抑制癌細(xì)胞增殖、抗血管形成及促上皮間質(zhì)轉(zhuǎn)化(epithelial to mesenchymal transitnio,EMT)發(fā)生等過程中的調(diào)控作用。方法:1建立裸鼠腹腔移植瘤模型:選擇高轉(zhuǎn)移潛能人肝癌細(xì)胞株MHCC-97H建立裸鼠皮下荷瘤模型,待荷瘤長至約1.5cm3,處死荷瘤裸鼠,取出腫瘤組織,將瘤塊剪成3mm直徑小塊供實(shí)驗(yàn)用。實(shí)驗(yàn)裸鼠分A組(實(shí)驗(yàn)組)和B組(對照組),每組6只,將預(yù)先處理好的實(shí)驗(yàn)用瘤組織各1塊埋入裸鼠腹腔建立裸鼠腹腔移植瘤模型。術(shù)后A組裸鼠每日索拉非尼30mg/kg(0.2ml)灌胃,日1次,B組給予等體積的溶劑,35天后處死所有裸鼠,取材。2對比索拉非尼治療組及對照組腫瘤的大小,免疫組織化學(xué)染色法檢測兩組腫瘤組織微血管密度(microvessel density,MVD)、內(nèi)源性缺氧標(biāo)記物HIF-1α(Hypoxia-inducible factor-1α)、E-鈣粘素(E-cadherin)及波形蛋白(Vimentin)的表達(dá)變化。3利用基因芯片技術(shù)檢測實(shí)驗(yàn)組及對照組mi RNA表達(dá)情況,并對其結(jié)果進(jìn)行生物信息學(xué)分析。Gene Ontology(GO)用來分析靶基因的分子功能。靶基因的顯著性信號通路利用Kyoto Encyclopedia of Genes and Genomes(KEGG)進(jìn)行分析。4通過生物信息學(xué)結(jié)果,分析并討論mi RNA在索拉非尼抑制癌細(xì)胞增殖、抗血管形成及促使EMT發(fā)生等過程所起的調(diào)控作用。結(jié)果:1 A、B兩組裸鼠腹腔成瘤率均為100%,實(shí)驗(yàn)期間兩組裸鼠均無死亡,未見明顯肉眼轉(zhuǎn)移。2實(shí)驗(yàn)組裸鼠腫瘤體積明顯小于對照組,MVD表達(dá)量明顯低于對照組,上述差異均有統(tǒng)計(jì)學(xué)差異(P0.05)。實(shí)驗(yàn)組HIF-1ɑ表達(dá)量高于對照組,兩組比較具備統(tǒng)計(jì)學(xué)差異(P0.05);與對照組相比,實(shí)驗(yàn)組E-cadherin的表達(dá)明顯下調(diào),而Vimentin表達(dá)上調(diào),兩組比較均有統(tǒng)計(jì)學(xué)差異(P0.05),提示索拉非尼促進(jìn)上皮間質(zhì)轉(zhuǎn)化的發(fā)生。3基因芯片分析mi RNA表達(dá)譜的差異發(fā)現(xiàn)28個mi RNAs在實(shí)驗(yàn)組及對照組中差異表達(dá),上調(diào)的mi RNAs有20個,下調(diào)的mi RNAs有8個。通過生物信息學(xué)分析結(jié)果判斷:mi R-762、mi R-150-5p明顯上調(diào),參與調(diào)控索拉非尼抑制肝癌細(xì)胞增殖;mi R-142a-3p雖然上調(diào)不明顯,但參與調(diào)控多個與腫瘤血管形成相關(guān)的生物學(xué)過程;上調(diào)的mi R-155-5p及下調(diào)的mi R-122-5p參與EMT的發(fā)生。結(jié)論:1索拉非尼具有抑制腫瘤生長及抗血管形成的雙重作用;并致腫瘤組織缺氧,誘導(dǎo)腫瘤上皮間質(zhì)轉(zhuǎn)化的發(fā)生。2 mi R-762、mi R-150-5p參與調(diào)控索拉非尼抑制肝癌細(xì)胞增殖的過程,mi R-142a-3p參與調(diào)控抗腫瘤血管形成過程。3 mi R-155-5p及mi R-122-5p參與調(diào)控索拉非尼誘導(dǎo)EMT的發(fā)生過程。
[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 偽 (Hypoxia-inducible factor-1 偽), 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.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R735.7

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1 宋曉;蔡振旭;;MicroRNAs的失調(diào)在高轉(zhuǎn)移肝細(xì)胞癌中的作用[J];中華臨床醫(yī)師雜志(電子版);2013年22期

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