【作者】 张燕；

【导师】 杨菁；

【作者基本信息】 武汉大学 ， 妇产科（专业学位）， 2015， 博士

【摘要】 目的：卵巢癌是一种临床发生率较高的妇科恶性肿瘤疾病,流行病学调查结果显示此疾病所造成的患者死亡率已位居各种妇科肿瘤疾病的首位,对妇女的身体健康及生命安全均造成十分严重的威胁。上皮细胞-间充质细胞转换(epithelial-mesenchymal transition, EMT)是一种细胞特殊分化过程,其在人体胚胎组织的正常发育过程中发挥着重要的生理作用,但同时也会诱发各种病理改变,近些年已逐渐引起较多研究人员的广泛关注。EMT在消化道肿瘤、皮肤癌、乳腺癌、卵巢癌、原发性肝细胞癌、胰腺癌和肺腺癌等多种类型癌症疾病的发生、发展过程中均发挥一定的生理作用,尤其可明显促进肿瘤组织的侵袭、远处转移等生理功能。EMT发生在肿瘤组织的微环境状态中,受到缺氧诱导因子-1α(HIF-1α)、转录因子Snail及黏附分子E-cadherin等较多细胞炎性因子的调节和控制,也是肿瘤组织和细胞发展和远处脏器组织转移的重要原因。本研究拟探讨EMT相关因子(HIF-1α、Snail、E-cadherin)在人粘液性卵巢癌细胞株3AO、人浆液性卵巢癌细胞株SKOV3、人透明细胞卵巢癌细胞株ES-2、人未分化卵巢癌细胞株TYK及卵巢癌组织中的表达及临床意义。方法：采用Boyden小室模型方法检测3AO、SKOV3、ES-2及TYK卵巢癌细胞的侵袭能力；采用western-blot和RT-PCR方法检测HIF-1α、Snail和E-cadherin在人卵巢癌细胞株3AO、SKOV3、ES-2和TYK中的表达情况；采用免疫组化检测HIF-α1、Snail和E-cadherin在卵巢癌组织中的表达情况：分析三种蛋白表达之间的相关性及与年龄、组织学分级、 FIGO分期、组织类型、肌层浸润、淋巴结转移等临床病理因素的关系。结果：1.人浆液性卵巢癌细胞株SKOV3、人透明细胞卵巢癌细胞株ES-2的侵袭细胞数分别为每高倍视野(132.6±6.4)个和(130.4±5.6)个,明显高于人未分化卵巢癌细胞株TYK(102.3±4.7)个和人粘液性卵巢癌细胞株3AO(104.1±5.2)个(P<0.01)；2. HIF-1α基因表达水平：SKOV3>ES-2>3AO>TYK; Snail基因表达水平：SKOV3=ES-2>3AO>TYK; E-cadherin基因表达水平：TYK>3AO>SKOV3= ES-2(P<0.05);3. H1F-1蛋白表达水平：SKOV3>ES-2>3AO>TYK; Snail蛋白表达水平：ES-2>SKOV3>3AO>TYK; E-cadherin蛋白表达水平；TYK=3AO>ES-2>SKOV3 (P<0.05);4.卵巢癌中HIF-1α阳性表达与组织学分级、组织类型和淋巴结转移明显相关(P<0.05); Snail阳性表达与手术病理分期、组织类型和淋巴结转移明显相关(P<0.05); E-cadherin阳性表达与组织类型、淋巴结转移明显相关(P<0.01)；5.卵巢癌中HIF-1α与Snail的表达呈正相关关系(r=0.231,P=0.021), Snail与E-cadherin的表达呈负相关关系(r=-0.225,P＝0.028)。结论：HIF-1α可能通过上调Snail的表达来抑制E-cadherin的表达,在卵巢癌的侵袭和转移中起重要作用。更多还原

【Abstract】 Objective:Ovarian cancer is a common malignant tumor of a female reproductive system with a high clinical incidence, and its mortality rate places in the top of gynecological malignancies causing a serious threat to women’s health and lives. Epithelial-mesenchymal conversion (epithelial-mesenchymal transition, EMT) is a special kind of cell differentiation, which plays an important physiological role in the normal development of human embryonic tissue, but it will also induce a variety of pathological changes and has in recent years gradually attracted more attention of researchers. EMT plays a role in many stages of development of a various cancers such as gastrointestinal cancer, skin cancer, breast cancer, ovarian cancer, hepatocellular carcinoma, pancreatic cancer and lung adenocarcinoma, in particular, EMT can promote invasion and metastasis of tumor tissue. EMT occurs in the tumor microenvironment being regulated and controlled by hypoxia-inducible factor-1α (HIF-1α), transcription factor Snail, E-cadherin adhesion and other inflammatory factors which are also the important reasons for development and distant organs metastasis of tumor tissues. This study aimed to investigate the expression and clinical significance of HIF-1α, Snail and E-cadherin in human mucinous ovarian cancer cell line 3AO, serous ovarian cancer cell line SKOV3, human clear cell ovarian cancer cell line ES-2 and human undifferentiated ovarian cancer cells strains the TYK and ovarian cancer tissues.Methods:Boyden chamber model was used to detect the invasion of 3AO, SKOV3, ES-2 and TYK ovarian cancer cells; western-blot and RT-PCR method were applied to detect the expression of HIF-1α, Snail and E-cadherin in human ovarian cancer cell line 3AO, SKOV3, ES-2 and TYK; immunohistochemistry was also used to analyze the expression of HIF-1α, Snail and E-cadherin in ovarian cancer tissue; correlation with the three proteins was analyzed as well as their relationship among age, histological grade, FIGO stage, histological type, myometrial invasion, lymph node metastasis and other clinicopathological factors.The results:1. The cell invasion numbers of human serous ovarian cancer cell line SKOV3, human clear cell ovarian cancer ES-2 were (132.6±6.4) and (130.4±5.6) per high-power field respectively, which were significantly higher than those of human undifferentiated ovarian cancer cell line TYK (102.3 ± 4.7) and human mucinous ovarian cancer cell line 3AO (104.1± 5.2) (P<0.01) per high-power field.2. Gene expression levels of HIF-1α:SKOV3> ES-2> 3AO> TYK; gene expression levels of Snail:SKOV3= ES-2> 3AO TYK; gene expression levels of E-cadherin: TYK> 3AO> SKOV3= ES-2 (P<0.05);3. Expression levels of HIF-la protein:SKOV3> ES-2> 3AO> TYK; expression levels of Snail protein:ES-2> SKOV3> 3AO> TYK; expression levels of E-cadherin protein; TYK= 3AO> ES-2> SKOV3 (P<0.05);4. The positive expression of HIF-la in ovarian cancer was significantly associated with histological grading, histological type and lymph node metastasis (P<0.05); Snail expression was significantly associated with surgical pathological staging, histological type and lymph node metastasis (P<0.05); E-cadherin positive expression was significantly associated with histological type and lymph node metastasis (P O.01);5. Expression of Snail was positively correlated with HIF-la in ovarian cancer (r= 0.231, P= 0.021), while Snail was negatively correlated with E-cadherin expression (r =-0.225, P= 0.028).In conclusion:HIF-la meybe inhibit the expression of E-cadherin by upregulating the expression of Snail, playing an important role in ovarian cancer invasion and metastasis.更多还原