【作者】 张然；

【导师】 王黎明；

【作者基本信息】 青岛大学 ， 妇产科学， 2018， 硕士

【摘要】 目的:探讨microRNA-145、SOX2和OCT4在原发性宫内膜样子宫内膜和卵巢双癌组织中的表达和意义,进而寻找更多支持原发性子宫内膜和卵巢双癌两部位癌组织同源性的证据。方法:1.收集20例原发性子宫内膜和卵巢双癌组织、10例转移性子宫内膜和卵巢双癌两部位癌组织中以及10例正常子宫内膜组织、10例正常卵巢组织标本,所有标本均为石蜡组织标本。2.采用荧光定量PCR法检测上述不同石蜡组织标本中的microRNA-145的表达情况,分析microRNA-145在双癌患者两部位癌组织中表达差异和相关性,分析microRNA-145的表达在原发性和转移性双癌不同组织中的表达的区别和联系。3.免疫组织化学二步法检测SOX2和OCT4蛋白在上述原发性子宫内膜和卵巢双癌组织、转移性子宫内膜和卵巢双癌组织中以及正常子宫内膜组织、正常卵巢组织中的表达情况,并将阳性表达的标本的免疫组化图片随机选取3幅用Image Pro-Plus软件进行定量分析。4.分析microRNA-145、SOX2和OCT4在不同组织的中表达的差异性及同一组织不同分子之间表达的相关性。结果:分别与相应的正常子宫内膜组织、正常卵巢组织作对比分析,microRNA-145、SOX2和OCT4在原发性、转移性宫内膜样子宫内膜和卵巢双癌两部位癌组织中均有异常表达。其中microRNA-145在子宫部位癌组织处(与正常子宫内膜组织相比)均呈低表达;在卵巢癌组织处(与正常卵巢组织相比)均呈高表达;SOX2和OCT4在双癌两部位癌组织中均呈阳性表达,正常子宫内膜组织、正常卵巢组织处均为阴性表达(其中仅一例正常卵巢组织中有小于5%的卵巢细胞呈浅棕色阳性表达,根据评分准则记为阴性表达)。统计学分析显示microRNA-145、SOX2和OCT4在原发性双癌两部位癌组织中的表达均无差异性(P均>0.05),且两处癌组织中SOX2和OCT4的表达均呈相关性(子宫内膜癌组织处统计值为:r=0.449,p=0.047,卵巢癌癌组织处统计值为r=0.449,P=0.047)。但是统计学分析显示原发性双癌两部位癌组织中的microRNA-145的表达与SOX2、OCT4均无明显的相关性(P均大于0.05)。转移性子宫内膜和卵巢双癌中两部位癌组织中上述三个分子的表达也均无差异性(P均>0.05),且在两处癌组织中也均发现了SOX2和OCT4的相关性表达,统计学值分别为子宫内膜癌组织:r=0.770,P=0.009,卵巢癌组织:r=0.716,P=0.020。同时通过原发性双癌和转移性双癌对比研究我们还发现原发性双癌组组织中的SOX2和OCT4的表达要明显低于转移性双癌(P<0.05),microRNA-145的表达则没有明显的差异性(P>0.05)。结论:研究结果提示microRNA-145、SOX2和OCT4可能均参与原发性、转移性宫内膜样子宫内膜和卵巢双癌两部位组织肿瘤的发生,且作用可能具有相似性,该结果与既往研究者研究结果一致,支持原发性子宫内膜和卵巢双癌的同源性,同时我们的研究结果还提示SOX2和OCT4可能在辅助正确区分原发性双癌和转移性双癌也有一定的指导意义。更多还原

【Abstract】 Objective: The aim of this study was to assess the involvements of SOX2,OCT4,and microRNA-145 in the tumorigenesis of double primary endometrioid endometrial and ovarian carcinomas(DPEEOCs).We also tried to find more clonal evidences for DPEEOCs.Methods: In our study,twenty DPEEOC patients were chosen.Ten metastatic double endometrioid endometrial and ovarian carcinomas(MDEEOCs)were also chosen.Ten normal endometrial tissues and ten normal ovarian tissues were also included.All samples were paraffin tissue specimens.The expression of microRNA-145 was detected by real-time quantitive PCR.We also analyzed the correlation and differences of microRNA-145 in the double two cancer sites of DPEEOC.Differences of microRNA-145 expression in DPEEOC and MDEEOC were also analyzed.Immunohistochemical staining(IHC)was used to measure the expression of OCT4 and SOX2 in DPEEOC tissues,MDEEOC tissues,and all normal tissues.As for those positively expressed tissues,three IHC pictures were chosen and quantitatively analyzed with Image Pro Plus.We also analyzed the differences of microRNA-145,SOX2 and OCT4 in different tissues.The correlation of microRNA-145,SOX2 and OCT4 in the same tissues were also analyzed.Results: The results showed that microRNA-145 was abnormal expressed in all DPEEOC and MDEEOC tissues.It was lower expressed in DPEEOC endometrial tissues while higher expressed in DPEEOC ovarian tissues compared to corresponding normal tissues.Both SOX2 and OCT4 were positively expressed in all cancer tissues,While they were negatively expressed in normal endometrial and ovarian tissues(one NOT tissue with faint yellow staining in less than 5% ovarian cells,while it was negtively expressed according to the standard).MicroRNA-145,SOX2,and OCT4 were expressed at similar levels in two cancer sites of a given DPEEOC or MDEEOC samples(all P > 0.05).We also found a positively correlation between the expression of SOX2 and OCT4 in both two cancer sites of DPEEOC samples;the statistical results in DPEEOC samples were as follows:(endometrial cancer tissues:r=0.449,p=0.047;ovarian cancer tissues: r = 0.449,P = 0.047).However,we did not found a correlation between the expression of microRNA-145 and SOX2 or OCT4(both P >0.05).SOX2 and OCT4 were also expressed in MDEEOC lesions;the statistical were as follows: endometrial cancer tissues,r = 0.770,P = 0.009;ovarian cancer tissues :r = 0.716,P = 0.020.Besides,MDEEOC sections expressed a higher level of SOX2 and OCT4 compared to corresponding DPEEOC tissues(P<0.05).However,no difference of microRNA-145 was found betweencorrespond cancer tissues of DPEEOC and metastic DEEOC(P > 0.05).Conclusion: We thought microRNA-145,SOX2,and OCT4 might play similar roles in the tumorigenesis of the two cancer sites in DPEEOCs.Our results were in accordance with other researchers’ reports about the clonality of DPEEOCs.Besides,we also found that SOX2 and OCT4 may have some implication in DPEEOC’ and MDEEOC’ diagnosis.更多还原