HE4在妇科恶性肿瘤早期诊断的价值及其表达缺失对肿瘤细胞增殖的影响

发布时间：2018-02-06 05:54

本文关键词： 卵巢癌子宫内膜癌人附睾蛋白4(HE4)　出处：《中国人民解放军医学院》2016年博士论文　论文类型：学位论文

【摘要】：研究背景：卵巢癌和子宫内膜癌是妇科常见恶性肿瘤,早期诊断、早期治疗、规范随访是改善预后的重要手段。CA125是目前应用最广泛的肿瘤标志物,但它在许多妇科良性疾病和内科疾病中也升高,用于诊断癌症的特异性不佳。HE4是由Kirchhoff于1991年从人的附睾远端上皮细胞中发现的,因其在恶性肿瘤组织中的特异性高表达正受到越来越多的关注。HE4作为卵巢癌血清标记物被认为要优于CA125,在子宫内膜癌早期诊断及预后判断方面也发挥重要的作用。目的：观察卵巢良性肿瘤、卵巢上皮性癌、子宫内膜癌患者血清和组织中CA1 25及HE4水平,并分析其在各组病例中的表达情况以及临床意义。通过RNA干扰技术观察HE4基因沉默对肿瘤细胞增殖、转移及侵蚀能力的影响。建立卵巢细胞系CaoV3裸鼠皮下移植瘤模型,体内观察HE4低表达对肿瘤生长的影响,以初步探求HE4与卵巢癌及子宫内膜癌发生的关系材料与方法：1、收集卵巢上皮性癌、良性上皮性卵巢肿瘤患者和子宫内膜癌患者术前血清,检测血清CA 125及HE4蛋白的表达水平,并分析其在肿瘤诊断中的灵敏度及特异度。2、收集卵巢上皮性癌、良性上皮性卵巢肿瘤患者和子宫内膜癌患者组织,检测组织中HE4蛋白的表达情况,分析其与肿瘤临床病理特征的相关性。3、构建shRNA-HE4慢病毒载体,转染到CaoV3细胞,观察HE4基因沉默对CaoV3增殖、转移及侵蚀能力的影响。4、建立CaoV3裸鼠皮下移植瘤模型,体内观察shRNA-HE4对卵巢癌肿瘤生长的影响。结果：1、血清HE4在卵巢癌高于健康对照组及良性肿瘤组,HE4诊断卵巢癌的特异度高于CA125。HE4水平与卵巢癌期别及淋巴转移相关。子宫内膜癌组HE4水平高于健康对照组及子宫内膜增生组,但与肿瘤分期等病理特征无相关性。2、组织中卵巢癌HE4表达阳性率高于卵巢良性肿瘤组,与卵巢癌期别及淋巴结转移相关。子宫内膜癌组HE4表达阳性率高于健康对照组及子宫内膜增生组,与淋巴结转移有关。3、成功构建HE4 shRNA慢病毒,并获得稳定表达的细胞株。敲低HE4基因后,细胞株增殖显著减慢,细胞周期出现G0/G1阻滞。4、HE4基因沉默能抑制肿瘤细胞在裸鼠体内的生长,抑制裸鼠肿瘤的体积,与对照组有显著性差异。结论：1、与CA125相比,HE4对于卵巢上皮性癌及子宫内膜癌有更好的诊断价值。2、HE4基因参与促进癌细胞的增殖、迁移、侵袭等行为。3、HE4基因沉默显著抑制卵巢癌细胞株在裸鼠体内的生长。
[Abstract]:Background: ovarian cancer and endometrial carcinoma are common malignant tumors in gynecology. Early diagnosis, early treatment and standardized follow-up are important means to improve prognosis. CA125 is the most widely used tumor marker. However, it is also elevated in many benign gynecological and internal diseases, and the poor specificity for the diagnosis of cancer. HE4 was found in human epididymal distal epithelial cells in 1991 by Kirchhoff. Because of its specific high expression in malignant tumor tissues, more and more attention has been paid to HE4 as a serum marker of ovarian cancer, which is considered to be superior to CA125. It also plays an important role in early diagnosis and prognosis of endometrial carcinoma. Objective: to observe benign ovarian tumor and epithelial ovarian carcinoma. The levels of CA1 25 and HE4 in serum and tissue of patients with endometrial carcinoma. The expression and clinical significance of HE4 gene silencing in each group were analyzed. The effect of HE4 gene silencing on tumor cell proliferation was observed by RNA interference technique. Establishment of ovarian cell line CaoV3 nude mice subcutaneous transplanted tumor model in vivo to observe the effect of low expression of HE4 on tumor growth. Methods: to investigate the relationship between HE4 and ovarian cancer and endometrial carcinoma. The serum samples were collected from patients with ovarian epithelial carcinoma, benign epithelial ovarian tumor and endometrial carcinoma before operation. The expression of CA125 and HE4 protein in serum was detected, and the sensitivity and specificity of CA125 and HE4 protein in the diagnosis of ovarian epithelial carcinoma were analyzed. The expression of HE4 protein in benign epithelial ovarian tumor and endometrial carcinoma was detected, and the correlation between the expression of HE4 protein and clinicopathological features of the tumor was analyzed. ShRNA-HE4 lentivirus vector was constructed and transfected into CaoV3 cells. The effect of HE4 gene silencing on the proliferation, metastasis and erosion of CaoV3 was observed. The effect of shRNA-HE4 on the growth of ovarian cancer was observed in vivo. Results the serum HE4 in ovarian cancer was higher than that in healthy control group and benign tumor group. The specificity of HE4 in diagnosis of ovarian cancer was higher than that of CA125.HE4, and the level of HE4 in endometrial carcinoma group was higher than that in healthy control group and endometrial hyperplasia group. The positive rate of HE4 expression in ovarian cancer was higher than that in benign ovarian tumors. The positive rate of HE4 expression in endometrial carcinoma group was higher than that in healthy control group and endometrial hyperplasia group, and was related to lymph node metastasis. HE4 shRNA lentivirus was successfully constructed and stably expressed cell lines were obtained. After knockout of HE4 gene, cell proliferation was significantly decreased and cell cycle was arrested by G _ 0 / G _ 1 arrest. HE4 gene silencing could inhibit the growth of tumor cells and tumor volume in nude mice, which was significantly different from that in control group. Conclusion: 1, compared with CA125, the silencing of HE4 gene can inhibit the growth of tumor cells in nude mice. HE4 has better diagnostic value for epithelial ovarian carcinoma and endometrial carcinoma. 2HE4 gene plays an important role in promoting the proliferation, migration and invasion of cancer cells. HE4 gene silencing significantly inhibited the growth of ovarian cancer cell line in nude mice.
【学位授予单位】：中国人民解放军医学院
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R737.3

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