【作者】 王娟娟；

【导师】 陈友国；

【作者基本信息】 苏州大学 ， 妇产科学（专业学位）， 2015， 硕士

【摘要】 目的:探讨FOXL2在正常宫颈组织与宫颈癌中的表达及MMP14干扰载体对FOXL2的调控及临床意义。方法:1.采用免疫组化法检测10例正常宫颈组织、18例宫颈癌组织(10例I期,8例II+III期)中FOXL2的表达情况,分析比较其表达与临床特征的关系。2.采用荧光定量PCR方法及western blot技术从基因和蛋白水平检测宫颈癌Hela细胞中MMP14、FOXL2的表达情况。3.MMP14质粒干扰载体转染宫颈癌Hela细胞,48小时后提取转染组及未转染组RNA,然后用q PCR方法检测转染前后FOXL2的m RNA相对表达量的变化。结果:1.FOXL2在正常宫颈组织中表达率为0%,而18例宫颈癌组织FOXL2阳性表达率为72.22%(13/18),其中宫颈癌I期阳性表达率50%,II期及III期阳性表达率为100%。FOXL2表达与宫颈癌临床分期、病理分级相关(P<0.05),而与宫颈癌患者的年龄、淋巴结转移无明显相关关系(P>0.05)。2.无论是m RNA水平,还是蛋白水平,在宫颈癌Hela细胞株中MMP14及FOXL2均表达。3.利用MMP14质粒干扰载体及脂质体2000转染宫颈癌Hela细胞48小时后转染组FOXL2的m RNA表达显著高于未处理组。结论:1.FOXL2与宫颈癌恶性度的相关性,可作为宫颈癌恶性程度的参考指标,对了解宫颈癌生物学行为、评估及预后具有一定的临床意义。2.在宫颈癌Hela细胞中,MMP14、FOXL2在基因水平及蛋白水平均表达,MMP14、FOXL2异常表达都是宫颈癌癌变及发展过程中的重要事件,提示多基因异常表达和相互作用可能是宫颈癌发生发展的根本原因。3.MMP14质粒干扰载体及脂质体2000转染宫颈癌Hela细胞后上调FOXL2的表达,但其具体信号通路及调控机制仍有待进一步研究,有可能为宫颈癌的分子诊断和基因靶向治疗提供新的理论依据,降低宫颈癌的术后复发率,提高其生存效率。更多还原

【Abstract】 Objectives:To explore the differential expression pattern of FOXL2 in normal cervical tissues and cervical cancer tissues and to investigate the clinical significance of effects of MMP14 sh RNA transfection on FOXL2 expression.Methods :1.In this study,we investigated FOXL2 expression in 10 normal cervical tissues and 18 cervical cancer tissues including 10 cases in stage I and 8 cases in stage II+III by immunohistochemical method,and analyzed the relationship between them.2.We estimated the expression of MMP14 and FOXL2 in cervical cancer cell line Hela from levels of gene and protein by fluorogenic quantitative PCR and Western Blot respectively.3.We constructed MMP14 sh RNA interference vector, incubating it with Lipo2000 and then transfected the mixture into Hela cell.48 hours after the interference,we extracted RNA and tested the relative expression of FOXL2 before or after our interference by q PCR.Results :1.We found no cases expressed FOXL2 in 10 normal cervical tissues,whereas there were 72.22%(13/18) cases expressed FOXL2 in cervical cancer tissues,including FIGO stage I cases with a ratio of 50% and FIGO stage II+III cases with a ratio of 100%. The outcome suggested that the expression of FOXL2 may be concerned with the clinical stages and pathological grading of cervical cancer(P<0.05), however,it showed no evident relevance with age and lymphatic metastasis(P>0.05).2.Cervical cancer cell line Hela expresses both MMP14 and FOXL2 from levels of gene and protein.3.48 hours after MMP14 sh RNA interference vector transfection,there was a obvious increase of FOXL2 expression,while the control group showed no evident changes.Conclusions:1.FOXL2 expression was relevant with cervical cancer malignancy, it may act as a indicator which would contribute to our understanding of the biological behavior of cervical cancer and to our assessment of clinical prognosis.2.Cervical cancer cell line Hela expressed both MMP14 and FOXL2 in gene and protein level.Their aberrant expression were distinctly important events during the canceration and progression of cervical cancer,it indicated that multi-gene mutation as well as their interaction with each other may be the fundamental pathogenesis of cervical cancer progression.3.The transfection of MMP14 sh RNA interference vector could enhance FOXL2 expression,but the specific signal pathway and regulatory mechanism need further investigation.FOXL2 may be a potential indicator of molecular diagnosis and target therapy for cervical cancer,it even helps to decrease recurrence ratio of cervical cancer after surgical therapy and increase survival rate.更多还原