Caspases are a family of cysteine protease enzymes composed of more than 10 members that play essential roles in apoptosis and inflammation. It has been reported that caspases play a critical role in regulating apoptosis at the maternal-conceptus interface in many species. However, the expression and regulation of caspases have not been determined in the endometrium in pigs. Therefore, we analyzed the expression, localization, and regulation of caspases in the endometrium during the estrous cycle and pregnancy in pigs. Real-time RT-PCR analysis showed that caspases were expressed in the endometrium during the estrous cycle and pregnancy. The expression of CASP6, CASP7, and CASP8 during the estrous cycle and CASP3, CASP6, CASP7, and CASP10 changed during pregnancy. Levels of CASP3 mRNA in the endometrium were higher on Day 12 of pregnancy than the estrous cycle and levels of CASP7 mRNAs were highest on Day 15 of estrous cycle and pregnancy. Immunohistochemistry analysis showed that CASP3 protein was localized to endometrial epithelial cells on Days 12 and 15 the estrous cycle and pregnancy, but cleaved CASP3 was localized only to luminal epithelial (LE) cells on Days 12 and 15 of pregnancy in the endometrium. CASP7 protein was localized to endometrial LE cells only on Day 15 of pregnancy. CASP3, CASP6, CASP7, CASP8, and CASP10 mRNAs were detectable in conceptus on D12 and D15 of pregnancy, and chorioallantoic tissues expressed CASP3, CASP6, CASP7, CASP8, CASP8, and CASP10 with increasing levels toward term pregnancy, except CASP3 mRNA. The effect of steroid hormones and interleukin-1βß (IL1B) on CASP3 expression and the effect of interferon-γ(IFNG) on CASP7 expression was determined by endometrial explant cultures and we found that CASP3 expression was increased by IL1B and CASP7 expression was increased by IFNG in a dose-dependent manner. These results showed that caspases were expressed in the endometrium during the estrous cycle and pregnancy in a stage- and/or pregnancy-specific dependent manner and some caspases were regulated by IL1B or IFNG in the endometrial tissues, suggesting that caspase may play an important role in regulating apoptosis for the establishment and maintenance of pregnancy at the maternal-conceptus interface in pigs.

Iris nertschinsk has been used generally as a decorative plant. However, it has been almost used as a medicine for therapy on various human diseases. In this study, we demonstrate the anti-tumor effect of Iris nertschinsk on human breast cancer cells. Firstly, we found that Iris nertschinsk dose-dependently induced cell death in human breast cancer cell lines, MCF7 and MDA-MB231. Moreover, phosphorylation of p53 was induced after Iris nertschinsk treatment in MCF7 cells, which has a functional p53, but not in MDA-MB231 cells, which has a dysfunctional p53. We next examined whether Iris nertschinsk induces caspase-dependent cell death. Caspase-7 was cleaved after Iris nertschinsk treatment in MCF7 cells. Interestingly, either caspase-3 or caspase-7 was cleaved in MDA-MB231 cells that p53 had been phosphorylated by Iris nertschinsk treatment, indicating that Iris nertschinsk induces apoptosis through the cleavage of caspase-3, -7 in human breast cancer cell lines, MCF7 and MDA-MB231, but related to the status of p53. Therefore, these results suggest that Iris nertschinsk could be used as a treatment for human breast cancer. This research is supported by National Institute of Agricultural Biotechnology research grant.