天津医药

天津医药 ›› 2017, Vol. 45 ›› Issue (7): 685-690.doi: 10.11958/20170345

• 细胞与分子生物学 • 上一篇    下一篇

生长抑素对子宫内膜癌分化程度的预测价值及对细胞生物学行为的影响

赵倩，周静怡，程媛，赵丽君，王甲琪，夏凤艳，王建六   

  1. 1 北京大学人民医院妇产科 （邮编 100044）； 2 唐山， 华北理工大学附属医院妇产科
  • 收稿日期:2017-03-17 修回日期:2017-05-27 出版日期:2017-07-15 发布日期:2017-08-08
  • 通讯作者: 王建六 E-mail:zhaoq128@163.com
  • 基金资助:
    国家自然科学基金资助项目 （81672571）

The effects of SST for predicting the differentiation of endometrial carcinoma and on cell biological behavior

ZHAO Qian,ZHOU Jing-yi,CHENG Yuan,ZHAO Li-jun,WANG Jia-qi,XIA Feng-yan,WANG Jian-liu   

  1. 1 Department of Obstetrics and Gynecology, Peking University People’ s Hospital, Beijing 100044, China; 2 Department of Obstetrics and Gynecology, North China University of Science and Technology Affiliated Hospital
  • Received:2017-03-17 Revised:2017-05-27 Published:2017-07-15 Online:2017-08-08

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摘要： 目的 探讨生长抑素（SST）基因在子宫内膜样腺癌组织及子宫内膜癌细胞系中的表达水平， 以及过表达该基因对子宫内膜癌 Ishikawa 细胞增殖及侵袭能力的影响。方法 选取正常子宫内膜、 子宫内膜样腺癌及子宫内膜浆液性癌组织切片， 采用免疫组织化学方法检测 SST 在各组织切片中的表达。收集手术新鲜标本， 病理诊断均为子宫内膜样腺癌， 按 2009 年 FIGO 标准病理分级， 其中高分化（G1） 7 例、 中分化（G2） 6 例、 低分化（G3） 5 例， 提取RNA， 采用实时荧光定量 PCR 方法检测 SST 的表达情况。选取 Ishikawa、 HEC-1A 及 KLE 细胞系， 采用实时荧光定量 PCR 及 Western blot 方法检测 SST 表达水平。以 SST 过表达慢病毒 pLVX-SST（Ish-SST 组） 及空载慢病毒 pLVX（Ish-ctr 组） 转染 Ishikawa 细胞， 荧光显微镜下观察转染效率。Western blot 检测两组 Ishikawa 细胞中 SST 蛋白表达水平。采用 CCK-8 及 Transwell 实验检测两组 Ishikawa 细胞的增殖及侵袭能力。结果 免疫组织化学结果显示， 与正常子宫内膜相比， SST 在子宫内膜样腺癌及子宫内膜浆液性癌中的表达显著增加。KLE 组 SST 的 mRNA 及蛋白水平较 Ishikawa 组及 HEC-1A 组高（P＜0.05）， HEC-1A 组 SST 的 mRNA 及蛋白水平高于 Ishikawa 组（P＜0.05）。在子宫内膜样腺癌中， 与 G1 和 G2 相比， SST 的表达在 G3 中显著增加 （P＜0.05）。在 Ishikawa 细胞传代 2 代后， IshSST 组 SST 蛋白表达水平显著高于 Ish-ctr 组。SST 过表达 Ishikawa 细胞增殖及侵袭能力与对照组差异无统计学意义（P＞0.05）。结论 SST 在低分化子宫内膜样腺癌中呈高表达； SST 过表达不能增加 Ishikawa 细胞的增殖及侵袭能力。

关键词: 子宫内膜肿瘤, 生长抑素, 细胞增殖, 肿瘤侵润

Abstract: Objective To investigate the expression levels of somatostatin (SST) gene in endometrial adenocarcinoma tissues and cell lines, and the effects of over-expression of SST gene on the proliferation and invasion of endometrial cancer cell line Ishikawa in vitro. Methods Tissue sections of normal endometrium, endometrioid adenocarcinoma and uterine papillary serous carcinoma were selected to detect the expressions of SST by immunohistochemical method. The total RNA was extracted from fresh specimens that were confirmed as endometrioid adenocarcinoma. According to FIGO staging,samples included G1 (7 cases), G2 (6 cases) and G3 (5 cases) of endometrioid adenocarcinoma. The SST expression levels were detected by real-time PCR. Three endometrial cancer cell lines, Ishikawa, HEC-1A and KLE, were selected and the expression levels of SST were detected by real-time PCR and Western blot assay. Transfection was performed with pLVXSST and pLVX. The transfection efficiency was observed by fluorescence confocal microscopy. The protein levels of SST were detected by Western blot assay. The assays of CCK- 8 and transwell were applied to examine variations in cell proliferation and invasion. Results Immunohistochemical results showed that SST expression was increased in endometrioid adenocarcinoma and uterine papillary serous carcinoma compared with that of normal endometrium. Real-time PCR showed that SST expression was significantly increased in G3 compared with that of G1 and G2 in endometrioid adenocarcinoma (P＜0.05). No matter mRNA or protein, SST levels were significantly increased in endometrial cancer cell line KLE compared with those of Ishikawa and HEC- 1A, and the expression levels of SST mRNA and protein were significantly increased in HEC- 1A group than those of Ishikawa group (P＜0.05). The expression of SST protein was significantly higher in the group of Ish- SST after 2 generations compared with that of Ish- ctr group. There were no significant differences in cell proliferation and invasive ability after over-expression of SST between Ishikawa cell group and control group (P＞0.05). Conclusion SST is highly expressed in poorly differentiated endometrial cancer cells. The proliferation and invasion are not increased after the over-expression of SST in Ishikawa cell line of endometrial cancer.

Key words: endometrial neoplasms, somatostatin, cell proliferation, neoplasm invasiveness

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