本文選題：孕裸鼠　切入點：宮頸癌　出處：《福建醫(yī)科大學(xué)》2014年碩士論文

【摘要】：目的 建立孕裸鼠宮頸癌皮下移植瘤模型，探討孕裸鼠宮頸癌移植瘤組織中MAPK信號通路活化水平及SPAG9（抗精子抗體）的表達變化及其相關(guān)性。 方法 選擇健康雌性裸鼠，交配成功后，隨機將孕裸鼠分成2組，實驗組10只，對照組10只，采用右腋皮下接種法。實驗組接種人宮頸癌HeLa細(xì)胞，對照組接種同體積生理鹽水，觀察孕裸鼠生長狀態(tài)及腫瘤的生長情況。接種14天后處死裸鼠，完整剝離腫瘤，HE染色觀察腫瘤細(xì)胞形態(tài)，采用蛋白免疫印跡（Western blot）技術(shù)檢測實驗組和對照組接種組織中SPAG9蛋白和MAPK信號轉(zhuǎn)導(dǎo)通路中ERK1/2、JNK1/2和P38磷酸化水平。采用實時熒光定量聚合酶鏈反應(yīng)（Real-time fluorescencequantitative PCR，，RQ-PCR）檢測實驗組和對照組接種組織中SPAG9mRNA的表達水平。 結(jié)果 1.孕裸鼠人宮頸癌HeLa細(xì)胞株單細(xì)胞懸液（2×106個/只）右腋皮下接種成瘤率100％。 2.實驗組皮下移植瘤組織中p-ERK1/2、p-JNK1/2、p-P38表達水平均高于對照組，差異均有統(tǒng)計學(xué)意義（P＜0.05）。 3.實驗組皮下移植瘤組織中p-ERK1/2與p-JNK1/2表達水平呈正相關(guān)（r=0.674，P＜0.05），均與p-P38表達無相關(guān)性（r分別為0.577、0.270，P＞0.05）。 4.實驗組皮下移植瘤組織中SPAG9mRNA相對表達量及SPAG9蛋白表達量均高于對照組，差異均有統(tǒng)計學(xué)意義（P＜0.05）。 5.實驗組皮下移植瘤組織中SPAG9mRNA相對表達量與p-ERK1/2、p-JNK1/2表達水平呈正相關(guān)（r分別為0.649、0.986，P＜0.05），與p-P38表達水平無相關(guān)性（r=0.312，P＞0.05）。實驗組皮下移植瘤組織中SPAG9蛋白表達量與p-ERK1/2、p-JNK1/2表達水平呈正相關(guān)（r分別為0.798、0.883，P＜0.05），與p-P38表達水平無相關(guān)性（r=0.317，P＞0.05）。 結(jié)論 1.孕裸鼠HeLa細(xì)胞皮下注射可成功建立孕裸鼠宮頸癌皮下移植瘤模型。 2.SPAG9基因可能主要通過參與JNK通路參與MAPK信號通路的調(diào)節(jié)。 3.SPAG9基因表達上調(diào)可能激活MAPK信號通路，從而參與孕裸鼠宮頸癌的發(fā)生發(fā)展。
[Abstract]:PurposeTo investigate the activation level of MAPK signaling pathway and the expression of anti-sperm antibody (SPAG9) in the transplanted tumor of cervical carcinoma in nude mice, the subcutaneous transplanted tumor model of cervical cancer in nude mice was established to study the relationship between the activation of MAPK signal pathway and the expression of anti-sperm antibody.MethodHealthy female nude mice were selected. After mating successfully, pregnant nude mice were randomly divided into two groups: the experimental group (n = 10) and the control group (n = 10).Human cervical cancer HeLa cells were inoculated in the experimental group and the control group were inoculated with the same volume of saline to observe the growth state and tumor growth of pregnant nude mice.After 14 days of inoculation, nude mice were killed, and tumor cells were observed by HE staining. The phosphorylation levels of SPAG9 protein and ERK1 / 2JNK1 / 2 and P38 in MAPK signal transduction pathway were detected by Western blot technique.Real-time fluorescencequantitative RQ-PCRR was used to detect the expression of SPAG9mRNA in the inoculated tissues of experimental group and control group.Result1.Single cell suspension of human cervical carcinoma HeLa cell line in pregnant nude mice (2 脳 106 / mouse) was inoculated subcutaneously in the right axilla and the tumorigenic rate was 100%.2.The expression of p-ERK1 / 2pJNK1 / 2p38 in subcutaneous transplanted tumor in the experimental group was higher than that in the control group (P < 0.05).3.There was a positive correlation between the expression of p-ERK1/2 and p-JNK1/2 in the subcutaneous transplanted tumor tissues in the experimental group (P < 0.05), but no correlation between the expression of p-P38 and p-ERK1/2 expression in the experimental group (P > 0.05).4.The relative expression of SPAG9mRNA and the expression of SPAG9 protein in the subcutaneous transplanted tumor in the experimental group were higher than those in the control group (P < 0.05).5.There was a positive correlation between the relative expression of SPAG9mRNA and the expression of p-ERK1 / 2 p-JNK1 / 2 in the subcutaneous transplanted tumor tissue in the experimental group (P < 0.05), and there was no correlation between the relative expression of SPAG9mRNA and the expression of p-ERK1 / 2 pJNK1 / 2 in the subcutaneous transplanted tumor (P < 0.05), but no correlation with the expression level of p-P38 (P > 0.05).Conclusion1.Subcutaneous injection of HeLa cells in pregnant nude mice can successfully establish subcutaneous transplanted tumor model of cervical cancer in pregnant nude mice.2.SPAG9 gene may be involved in the regulation of MAPK signaling pathway by participating in JNK pathway.Upregulation of 3.SPAG9 gene expression may activate MAPK signaling pathway and thus participate in the occurrence and development of cervical cancer in pregnant nude mice.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R737.33

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