人子宫肌瘤细胞分离培养及其体外雌孕激素受体的表达  

Isolation and culture of hunman uterine leiomyoma cells in vitro and expression of estrogen and progesterone receptor

作　　者：张春华[1] 朱勇[2] 温泽清[1] 

机构地区：[1]山东大学附属省立医院妇产科,山东济南250021 [2]山东中医药大学附属医院普外科,山东济南250011

出　　处：《中华肿瘤防治杂志》2010年第21期1748-1751,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：山东省自然科学基金(Y2006C67)

摘　　要：目的:探讨一种短时间内在体外获得大量子宫肌瘤细胞的方法,并对其进行生物学特性鉴定。方法:应用胶原酶消化法分离获得人子宫肌瘤细胞,体外培养扩增,免疫组织化学法对肌瘤细胞进行鉴定,并比较雌孕激素受体(estrogen recep-tor,ER;progesterone receptor,PR)在体外培养的肌瘤细胞、肌瘤组织及正常平滑肌组织中的表达情况。结果:应用胶原酶消化法体外分离培养并获得大量的纯度较高的子宫肌瘤细胞;镜下观察,体外培养生长良好的子宫肌瘤细胞呈梭形或不规则三角形,呈放射状或旋涡状生长;RT-PCR结果显示,与同源子宫肌瘤组织相比(ER0.91±0.30;PR 0.60±0.11),培养4代的肌瘤细胞ER(0.92±0.24),PR(0.62±0.14)的表达水平无显著差异(P>0.05),而与正常平滑肌组织(ER 0.64±0.31;PR 0.46±0.16)相比则有显著提高,P<0.05。结论:成功建立人子宫肌瘤细胞有限细胞系,且在体外培养的子宫肌瘤细胞和人体内肌瘤细胞相似的生物活性,为子宫肌瘤的研究提供了快速有效地实验模型。OBJECTIVE：To study is how to get the uterine leiomyoma cells in vitro which have the similar bionomics to the leiomyoma cells in human body,and is to discuss the senses of ER,PR expressions in the uterine leiomyoma tissues,normal autologous unaffected myometrium and the cultured uterine leiomyoma cells in vitro.METHODS：Human uterine leiomyoma tissues were dissected and cultured.Characterization of the cultured cells was examined using immunostaining with monoclonal antibodies to a muscle-specific protein desmin.RT-PCR was used to detect the mRNA expression of ER,PR in the cultured leiomyoma cells,uterine leiomyoma tissues and the normal autologous unaffected myometrium.RESULTS：The uterine leiomyoma cells were got by using collagenase dissection and were confirmed by immunostaining with monoclonal antibodies to a muscle-specific protein desmin.RT-PCR analysis revealed that ER,PR mRNA expressions have no significant changes between the cultuled cells（ER 0.92±0.24;PR 0.62±0.14） and the leiomyoma tissues（ER 0.91±0.30;PR 0.60±0.11;P0.05）,however ER,PR mRNA expressions in the normal autologous unaffected myometrium（ER 0.64±0.31;PR 0.46±0.16） were decreased to compare with the cultured cells and the tissues（P0.05）.CONCLUSION：The uterine leiomyoma cells can be got through the method of collagenase dissection which have the similar bionomics to the leiomyoma cells in human body in the genes of ER,PR expression.

关 键 词：子宫肌瘤细胞 雌激素受体 孕激素受体 体外 培养 

分 类 号：R737.33[医药卫生—肿瘤]