【作者】 张慧；

【导师】 李巧云；

【作者基本信息】 郑州大学 ， 妇产科， 2015， 硕士

【摘要】 研究背景子宫内膜癌是常见的妇科恶性肿瘤之一,以来源于子宫内膜癌腺体最常见,近年发病率在世界范围内呈上升趋势[3],部分患者因合并症及自身原因不能耐受手术或放疗、化疗治疗,临床医生一直试图寻求一种既能产生疗效又能提高生活质量的治疗方案。人肌母细胞基因疗法(HMGT)作为目前世界医疗水平较前研的治疗手段,可修复退化肌纤维受损基因补充活细胞。柳叶刀在1990年7月14日公布它是世界上第一个人类基因治疗,其可通过修复受损基因补充Duchenne型肌营养不良症的抗肌萎缩蛋白[13]。据来自美国食品药品管理局批准的II/III期临床试验中的数据表明,肌母细胞移植治疗肌营养不良症有明显的的疗效和安全性的优点;由FDA和EMA批准的II/III期缺血性心肌病临床试验中表现出显著的的疗效和较高的安全性[24];研究表明在II型糖尿病中肌母细胞参与了胰岛素信号通路和线粒体生物合成的功能[66],可以调节基因的表达水平;近年来,肌母细胞对胃癌动物模型和前列腺癌根治术后尿失禁也有一定的治疗作用。实验研究显示,肌母细胞移植入宿主细胞后可以导致肿瘤细胞的周期停滞与诱导凋亡,在前列腺癌小鼠模型中[65],注射肌母细胞能够抑制实体肿瘤的生长及转移,从而达到肿瘤的治疗与防治的目标。研究目的建立裸鼠子宫内膜癌模型,研究肌母细胞和子宫内膜癌细胞在裸鼠体内的相互作用,探讨肌母细胞对子宫内膜癌的初步治疗作用,为靶向子宫内膜癌的治疗提供理论与科学依据。如果该实验研究达到预期效果,将进一步进入临床研究阶段,并在子宫内膜癌的治疗中取得突破性的进展,人肌母细胞基因疗法成为了继手术、放疗、化疗等治疗手段之外的新的治疗方法,不仅能够有效的延长子宫内膜癌患者的长期生存率,而且可以极大的提高肿瘤患者的生活质量。材料与方法建立子宫内膜癌裸鼠模型,并将构建成功的30个裸鼠模型随机分为3组,即实验组、对照组、化疗组,每组10只,三组分别给予肌母细胞瘤体注射、腹腔注射顺氯氨铂、腹腔注射生理盐水。注射后每两日测量肿瘤长、短径,观察肿瘤及裸鼠变化,连续观察4周,计算各组肿瘤体积、抑瘤率及裸鼠体重及精神变化。HE染色观察各组肿瘤组织形态学改变,免疫组织化学法检测肿瘤中PTEN蛋白的表达水平。研究结果实验前对照组,化疗组及实验组裸鼠体重、肿瘤体积分别为(17.16±0.73)g、(38.27±2.5 8)mm3,(17.02±0.40)g、(35.99±1.32)mm3,(17.05±0.38)g、(37.05±2.9 3)mm3,实验后三组裸鼠体重,肿瘤体积分别为(16.23±0.3 1)g、(1 5 4.0 2±1 3.6 6)mm3,(17.26±0.50)g、(54.94±10.84)mm3,(19.50±0.49)g、(21.64±7.56)mm3;实验组、化疗组抑瘤率分别为85.95%、64.33%;三组PTNE阳性率分别为30%、40%、60%;HE染色光镜下观察,对照组可见大量典型的肿瘤细胞、异型性明显;实验组肿瘤细胞固缩、胞核裂解,可见大量凋亡坏死细胞;化疗组可见瘤体部分局限坏死组织,瘤体仍存在肿瘤细胞。结论1.实验组瘤体明显缩小,瘤体生长较化疗组缓慢,提示肌母细胞可能成为临床上子宫内膜癌的有效治疗方法。2.实验组瘤体HE染色提示癌细胞大量裂解坏死,免疫组化提示PTEN高表达,其治疗效果较化疗组明显。3.肌母细胞治疗裸鼠子宫内膜癌时对小鼠体重的影响是正向的,在抑制肿瘤增殖的同时,对体重、食欲、精神等方面的影响较小,可上调PTEN表达。4.肌母细胞较顺氯氨铂可更好地抑制裸鼠子宫内膜癌增殖,应用于子宫内膜癌治疗方面有一定可行性。更多还原

【Abstract】 Research BackgroundEndometrial carcinoma is a group of neoplasms occurred in endometrial epithelial tissue, most of which come from endometrial glands. So common in gynecological cancer in recent years, the incidence rate in the world is rising rapidly, some patients with complications can not tolerate surgery or radiation therapy, chemotherapy,in order to overcome this dilemma, clinicians have been trying to find a treatment, which not only works as we hope but also enhances the quality of life. Human myoblasts gene therapy(HMGT) can repair muscle fiber degeneration and complement genes in living cells, as the most advanced method in the world’s medical research over the previous level. In the July 14, 1990 the article published in The Lancet claimed HMGT was the Dystroph that the world’s first human gene therapy for Duchenne muscular dystrophy through repairing impaired genes. According to the data, which comes from the phase II/Ⅲ clinical trials approved by the US Food and Drug Administration, showed that myoblast transplantation in the treatment of muscular dystrophy has obvious advantages in efficacy and safety. Approved by the FDA and EMA Phase II / III clinical trials in ischemic cardiomyopathy exhibited significant efficacy and safety. Research shows that myoblast involved in insulin signaling pathways and mitochondrial biogenesis, in type II Diabetes,what’s more,it can still adjust the level of gene expression. In recent years, researches confirm myoblasts have significant therapeutic effects in the treatment of stomach cancer in animal models and urinary incontinence after radical prostatectomy.Co-culture experiments showed that the proliferation and differentiation of myoblasts can induce tumor cell cycle arrest and apoptosis in prostate cancer mouse models and injection of myoblasts can inhibit the growth and metastasis of solid tumors. Research ObjectiveTo establish nude mice endometrial carcinoma animal model, to study the interaction of myoblasts and endometrial cancer cells in nude mice, Simple to investigate the treatment of myoblasts cell on nude mice endometrial carcinoma, to provide theoretical and scientific basis for targeted therapy of endometrial carcinoma.If the basis research’s results as hoped at the beginning, the further clinical studies will be performed, and will be a breakthrough progress in the treatment of endometrial carcinoma, which will be another therapeutic means to the treatment of endometrial carcinoma beyond surgery, radiotherapy, chemotherapy, the members’ quality of life in cancer patients wil greatly improved. Research MethodsTo establish nude mice endometrial carcinoma animal model,then the nude mice were randomly divided into Experiment group, Control group and Chemotherapy group, with 10 rats in each group.the Experiment group was given intratumoral injection of myoblasts cell, the Control group was given intratumoral injection of normal saline and the Chemotherapy group was given intraperitoneal injection of Cis-platinum. Measured the neoplasm long axis and short axis once two days after the injection for 4 weeks, observed the change of neoplasm and nude mice for 4 weeks. Calculated the neoplasm volume, neoplasm inhibition rate and body weight change of nude mice. The morphology change of neoplasm tissue specimens was observed by HE Staining light microscope method. The expression of PTNE of neoplasm tissue specimens was tested by immunohistochemical detection. ResultsBefore the experiment,the nude mice weight and the neoplasm volume of Control group and Chemotherapy group, Experiment group were respectively(17.16±0. 7 3) g,( 38.27±2. 5 8) mm3,( 17.02±0. 4 0) g,( 35.99±1. 3 2) mm3,( 17.05±0. 3 8) g,( 37.05±2. 9 3) mm3. After the experiment, the nude mice weight and the neoplasm volume of the three groups were respectively( 1 6. 2 3 ±0. 3 1) g,( 1 5 4. 0 2 ±1 3. 6 6) mm3,(1 7. 2 6 ±0. 5 0)g,( 5 4. 9 4 ±1 0. 8 4) mm3,(1 9. 5 0 ±0. 4 9)g,(2 1. 6 4 ±7. 5 6) mm3. The neoplasm inhibition rate of the Experiment group and the Chemotherapy group were respectively 8 5. 9 5 %, 6 4. 3 3 %. T he positive rate of PTNE in the three groups were respectively 30%, 40%, 60%. In the Control group, the typical neoplasm cells and atypia cells were observed by HE Staining light microscope method. In the experiment group, the phenomenon of the cellular pyknosis, karyorrhexis and lots of apoptosis necrosis cells was obviously observed by the light microscope. Little of necrosis tissue was observed in the Chemotherapy group. Research Conclusions1. In the Experimental group, the neoplasm volume was significantly reduced and thespeed of growth In the experimental was slower what in Chemotherapy group, suggesting that myoblasts may be an effective clinical treatment for endometrial carcinoma.2. The morphology change of neoplasm tissue specimens was observed by HE Staining light microscope method,in the experimental group, lots of tumor lysis could be observed. The expression of PTNE of neoplasm tissue specimens was tested by immunohistochemical detection,and the expression of PTEN was more easily detected.suggesting that the experimental group was better than the Chemotherapy group in the clinical treatment for endometrial carcinoma.3. The influence of myoblast in the treatment of nude mice’ s endometrial carcinoma on body weight is positive, while in the inhibition of tumor growth, the impact on body weight, appetite, mental and other aspects is the lower. What’s more, the up-regulated expression of PTEN can be observed in the experimental group.4. The myoblasts cell and Cis-platinum can both inhibit the proliferation of endometrial carcinoma in nude mice, and the myoblasts cell is more effective. The therapy of myoblasts cell applied to the endometrial carcinoma is feasible.更多还原