MAPK信号通路及SPAG9在孕裸鼠宫颈癌皮下移植瘤的表达水平的研究

发布时间：2018-04-02 16:33

本文选题：孕裸鼠　切入点：宫颈癌　出处：《福建医科大学》2014年硕士论文

【摘要】：目的建立孕裸鼠宫颈癌皮下移植瘤模型，探讨孕裸鼠宫颈癌移植瘤组织中MAPK信号通路活化水平及SPAG9（抗精子抗体）的表达变化及其相关性。方法选择健康雌性裸鼠，交配成功后，随机将孕裸鼠分成2组，实验组10只，对照组10只，采用右腋皮下接种法。实验组接种人宫颈癌HeLa细胞，对照组接种同体积生理盐水，观察孕裸鼠生长状态及肿瘤的生长情况。接种14天后处死裸鼠，完整剥离肿瘤，HE染色观察肿瘤细胞形态，采用蛋白免疫印迹（Western blot）技术检测实验组和对照组接种组织中SPAG9蛋白和MAPK信号转导通路中ERK1/2、JNK1/2和P38磷酸化水平。采用实时荧光定量聚合酶链反应（Real-time fluorescencequantitative PCR，，RQ-PCR）检测实验组和对照组接种组织中SPAG9mRNA的表达水平。结果 1.孕裸鼠人宫颈癌HeLa细胞株单细胞悬液（2×106个/只）右腋皮下接种成瘤率100％。 2.实验组皮下移植瘤组织中p-ERK1/2、p-JNK1/2、p-P38表达水平均高于对照组，差异均有统计学意义（P＜0.05）。 3.实验组皮下移植瘤组织中p-ERK1/2与p-JNK1/2表达水平呈正相关（r=0.674，P＜0.05），均与p-P38表达无相关性（r分别为0.577、0.270，P＞0.05）。 4.实验组皮下移植瘤组织中SPAG9mRNA相对表达量及SPAG9蛋白表达量均高于对照组，差异均有统计学意义（P＜0.05）。 5.实验组皮下移植瘤组织中SPAG9mRNA相对表达量与p-ERK1/2、p-JNK1/2表达水平呈正相关（r分别为0.649、0.986，P＜0.05），与p-P38表达水平无相关性（r=0.312，P＞0.05）。实验组皮下移植瘤组织中SPAG9蛋白表达量与p-ERK1/2、p-JNK1/2表达水平呈正相关（r分别为0.798、0.883，P＜0.05），与p-P38表达水平无相关性（r=0.317，P＞0.05）。结论 1.孕裸鼠HeLa细胞皮下注射可成功建立孕裸鼠宫颈癌皮下移植瘤模型。 2.SPAG9基因可能主要通过参与JNK通路参与MAPK信号通路的调节。 3.SPAG9基因表达上调可能激活MAPK信号通路，从而参与孕裸鼠宫颈癌的发生发展。
[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.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33

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