Department of Obstetrics and Gynecology, The Catholic University of Korea, Seoul, Republic of Korea.
Int J Oncol. 2012 Jun;40(6):2013-21. doi: 10.3892/ijo.2012.1370. Epub 2012 Feb 14.
This study aimed to analyze expression of Müllerian inhibiting substance type II receptor (MISRII) protein and mRNA in cervical neoplasia, to demonstrate the growth inhibition of cervical cancer cells by administration of highly purified recombinant human Müllerian inhibiting substance (MIS) and, furthermore, to evaluate the clinical significance of MIS as a biological modifier for MIS receptor expressing tumors. Reverse transcriptase polymerase chain reaction (RT-PCR) was used for MISRII mRNA expression, and in situ hybridization and immunohistochemistry were used to observe expression, location of MISRII mRNA and protein, respectively. To demonstrate the effect of MIS on the viability of cervical cancer cells, methyl thiazole tetrazolium (MTT) assay was performed. Flow cytometry was used to evaluate the cell cycle distribution after exposure to MIS in cervical cancer cells, and the annexin-V-FITC staining method was performed to demonstrate apoptosis by MIS in cervical cancer cells. Expression of MISRII protein and mRNA were observed in all normal cervical and cervical carcinoma tissues. There was no significant difference in expression of MISRII protein and MISRII mRNA between normal cervical and cervical carcinoma tissues. MTT assay showed negative correlation between MIS exposure time and the viability of cervical cells (P=0.008). The changes in cell cycle distribution after MIS exposure suggest that MIS plays an important role in inducing cellular apoptosis by causing arrest at the G1 phase and increasing cells at sub-G0G1 phase. Annexin-V-FITC staining methods showed that cellular apoptosis was, respectively, 10.44 and 12.89% after 24 and 48 h of MIS exposure in cervical carcinoma cells. There was a negative correlation between cellular survival and MIS exposure time. This study demonstrates that MISRII is present on normal cervical and cervical carcinoma tissues, and MIS shows receptor-mediated antiproliferative effect on cervical cells in vitro. These data suggest that MIS may be used as a biological modifier or therapeutic modulator on MISRII-expressing tumors in the future.
本研究旨在分析 Müllerian 抑制物质 II 型受体（MISRII）蛋白和 mRNA 在宫颈癌中的表达，通过给予高纯度重组人 Müllerian 抑制物质（MIS）来证明其对宫颈癌细胞的生长抑制作用，并进一步评估 MIS 作为表达 MIS 受体的肿瘤的生物调节剂的临床意义。逆转录聚合酶链反应（RT-PCR）用于 MISRII mRNA 表达，原位杂交和免疫组织化学分别用于观察 MISRII mRNA 和蛋白的表达、定位。为了证明 MIS 对宫颈癌细胞活力的影响，进行了甲基噻唑四唑（MTT）测定。流式细胞术用于评估暴露于 MIS 后宫颈癌细胞的细胞周期分布，并用 Annexin-V-FITC 染色法证明 MIS 诱导的宫颈癌细胞凋亡。在所有正常宫颈和宫颈癌组织中均观察到 MISRII 蛋白和 mRNA 的表达。正常宫颈和宫颈癌组织中 MISRII 蛋白和 MISRII mRNA 的表达无显着差异。MTT 测定显示，MIS 暴露时间与宫颈细胞活力之间呈负相关（P=0.008）。MIS 暴露后细胞周期分布的变化表明，MIS 通过导致 G1 期停滞和增加亚 G0G1 期细胞，在诱导细胞凋亡中起重要作用。Annexin-V-FITC 染色法显示，宫颈癌细胞在 MIS 暴露 24 和 48 小时后，细胞凋亡分别为 10.44%和 12.89%。细胞存活率与 MIS 暴露时间之间存在负相关。本研究表明，MISRII 存在于正常宫颈和宫颈癌组织中，MIS 在体外对宫颈细胞表现出受体介导的抗增殖作用。这些数据表明，MIS 将来可能被用作表达 MISRII 的肿瘤的生物调节剂或治疗调节剂。
