Zhao Shu Jun, Chen Xiao Jun, Feng You Ji
Obstetrics and Gynecology Hospital of Fudan University, Shanghai 200011.
Fen Zi Xi Bao Sheng Wu Xue Bao. 2007 Apr;40(2):103-12.
The study was undertaken to investigate the effect of long-term treatment with MPA on the growth and invasiveness of endometrial carcinoma Ishikawa cells and the expression of the subtype of estrogen receptor and progesterone receptor. The human endometrial carcinoma Ishikawa cells were continually exposed to 2.5 micromol/L step-wise increase of MPA. MTT assay, flow cytometry and Matrigel invasion assay were used to detect the effect of MPA on the growth, cell cycle distribution and the invasion capability of the cells respectively. RT-PCR was performed to detect the changes of CyclinD1, MMP2 and MMP9 over different time treatment of MPA. Immunocytochemistry was used to examine the expression of estrogen receptor subtype and progesterone receptor subtype. Endometrial carcinoma Ishikawa cells became resistant to the inhibitory effect of MPA over ten months MPA treatment. The cells resistant to the growth inhibitory effect were named progestin-resistant Ishikawa cells and the cells which the progestin-resistant originated from were named the parent Ishikawa cells. The effects of MPA shifted from growth and invasiveness inhibitory effects on the parent Ishikawa cells to stimulatory effect on the progestin-resistant Ishikawa cells. Consistent with the phenomena, the treatment with MPA on Ishikawa cells resulted in statistically significant decreases of CyclinD1, MMP2 and MMP9 gene expression, reversely, the treatment with MPA on the progestin-resistant Ishikawa cell resulted in statistically significant increases of CyclinD1, MMP2, MMP9 gene expression. Immunocytochemical analysis showed reduced ERalpha and PR-B expression and increased ERbeta expression in the progestin-resistant Ishikawa cells compared with parental Ishikawa cells. From the experiment, it was concluded that the prolonged treatment with MPA on Ishikawa cell could give rise of the resistant effect of MPA, the effect of MPA on the growth and invasiveness capability of endometrial carcinoma shifted from inhibitory to stimulatory. The imbalance of ER subtype and PR subtype might contribute to the mechanisms involved in the progesterone resistance over long-term MPA treatment.
本研究旨在探讨甲羟孕酮（MPA）长期治疗对子宫内膜癌Ishikawa细胞生长、侵袭以及雌激素受体和孕激素受体亚型表达的影响。将人子宫内膜癌Ishikawa细胞持续暴露于浓度以2.5 μmol/L逐步递增的MPA中。分别采用MTT法、流式细胞术和基质胶侵袭实验检测MPA对细胞生长、细胞周期分布及侵袭能力的影响。通过RT-PCR检测MPA不同时间处理后细胞周期蛋白D1（CyclinD1）、基质金属蛋白酶2（MMP2）和基质金属蛋白酶9（MMP9）的变化。采用免疫细胞化学法检测雌激素受体亚型和孕激素受体亚型的表达。经10个月的MPA处理后，子宫内膜癌Ishikawa细胞对MPA的抑制作用产生耐药。将对生长抑制作用耐药的细胞命名为孕激素耐药Ishikawa细胞，将产生孕激素耐药的起源细胞命名为亲代Ishikawa细胞。MPA对亲代Ishikawa细胞的作用从生长和侵袭抑制转变为对孕激素耐药Ishikawa细胞的刺激作用。与这些现象一致，MPA处理Ishikawa细胞导致CyclinD1、MMP2和MMP9基因表达在统计学上显著降低，相反，MPA处理孕激素耐药Ishikawa细胞导致CyclinD1、MMP2、MMP9基因表达在统计学上显著增加。免疫细胞化学分析显示，与亲代Ishikawa细胞相比，孕激素耐药Ishikawa细胞中雌激素受体α（ERα）和孕激素受体B（PR-B）表达降低，雌激素受体β（ERβ）表达增加。从实验得出结论，MPA对Ishikawa细胞的长期处理可导致MPA耐药，MPA对子宫内膜癌生长和侵袭能力的作用从抑制转变为刺激。雌激素受体亚型和孕激素受体亚型的失衡可能是长期MPA治疗导致孕激素耐药的机制之一。
