Bone marrow mesenchymal stem cells (MSCs) are considered a potential cell source for stem cell-based bone tissue engineering. However, noticeable limitations of insufficient supply and reduction of differentiation potential impact the feasibility of their clinical application. This study investigated the in vitro function of steroids and gender differences on the proliferation and differentiation of rat MSCs. Bone marrow MSCs of age-matched rats were exposed to proliferation and osteogenic differentiation media supplements with various concentrations of 17β-estradiol (E2) and dexamethasone. Cell proliferation was measured by MTS assay; osteogenic markers and steroid-associated growth factors and receptors were evaluated by ELISA and real-time PCR. The results revealed that supplements of E2 and dexamethasone increase MSC proliferation in a biphasic manner. The optimal dose and interaction of steroids required to improve MSC proliferation effectively varied depending on the gender of donors. Supplementation of E2 effectively improves osteogenic differentiation markers including ALP, osteocalcin and calcium levels for MSCs isolated from both male and female donors. The mRNA of TGF-β1 and BMP-7 are also up-regulated. However, effective doses to maximally improve osteogenic potentials and growth factors for MSCs are different between male and female donors. The relationship between steroid receptors, osteogenic markers and cytokines are also varied by genders. The outcomes of the present study strongly indicate that steroids potentially function as an effective modulator to improve the capacity of MSCs in bone regeneration. It provides crucial information for improving and optimizing MSCs for future clinical application of bone regeneration.

Human bone marrow mesenchymal stromal cells (MSCs) with self-renewal and multiple differentiation potentials are considered a possible cell source for tissue engineering and regenerative medicine. However, the limited amount of MSCs in bone marrow and the loss of differentiation capacity following in vitro expansion restrict their practical application. Effective improvement of MSC proliferation is necessary for the clinical application of MSC-based tissue engineering. The effects of estrogen supplements on proliferation and characterizations of human MSCs were investigated at the present study. Supplements of 17-β estradiol (E2) significantly increase the proliferation of human MSCs in vitro. The dose range of E2 to significantly increase MSC proliferation differs in the gender of MSC donor. E2 supplementation in cell proliferation maintains characterizations of MSCs, including cell surface markers, and osteogenic and adipogenic differentiation capacities. These data indicate that estrogen treatment can play an important role in improving human MSCs' expansion in vitro, which will effectively facilitate MSCs' function in the practical application of tissue engineering and regeneration.

We conducted the present study to determine the chemoresistance mechanisms in clear cell carcinoma of the ovary (CCC). Five human CCC cell lines (HAC-2, RMG-I, RMG-II, KK, and KOC7c) were used in this study. The sensitivity of the cells to the anticancer agents was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and we assessed drug sensitivity by calculating assay area under the curve (AUC) for each agent. The expression of multi-drug resistance genes (MDR-1, MRP-1, MRP-2) was detected by reverse transcriptionpolymerase chain reaction (RT-PCR). Glutathione (GSH) concentration was measured by an enzymatic assay. Topoisomerase (topo) I activity was assayed in terms of relaxation of supercoiled plasmid substrate DNA. The IC 50 to anticancer agents ranged widely. The assay AUC indicated that 3 of 5 cell lines (RMG-I, RMG-II, and KK) were sensitive to paclitaxel (PTX), 3 (HAC-2, RMG-I, and RMG-II) were sensitive to 7-ethyl-10-hydroxycamptothecin (SN-38), which is an active metabolite of camptothecin (CPT-11), and only one (HAC-2) was sensitive to cisplatin (CDDP). All cell lines were resistant to mitomycin-C (MMC) and etoposide (VP-16). The MRP-1 gene was detected in all cell lines. Only one cell line showed both MRP-2 and MDR-1 gene expression. Except for HAC-2 cells, expression of MRP genes was related to CDDP resistance, and MDR-1 gene expression was associated with PTX resistance. GSH concentrations increased after exposure to CDDP or MMC in all cell lines. There was a significant correlation between topo-I enzymatic activity and the response to SN-38. The present study revealed several resistance mechanisms in CCC and the results suggested that PTX and CPT-11 might be effective agents to treat CCC.

The prognostic significance of PTEN expression in endometrial carcinoma has not been clear. We conducted the present study to clarify the relationship between PTEN expression and prognosis in advanced endometrial carcinoma. 4 showed that PTEN ϩ/-mice over 6 months old frequently developed endometrial carcinoma. These findings suggest that PTEN is strongly involved in the development and/or progression of endometrial carcinoma.Studies have also shown the relationship between PTEN and prognosis in several cancers, including endometrial carcinoma. However, these results are controversial. In 2 studies, loss of PTEN expression was related to poor prognosis in glioblastoma and prostate cancer. 5,6 In contrast, PTEN mutation was associated with favorable survival in endometrial carcinoma. 7 Two issues make it difficult to evaluate PTEN as a prognostic factor in endometrial carcinoma. First, because about 75% of patients with endometrial carcinoma present with stage I disease, which has a good prognosis, 8 a large number of advanced cases are needed to analyze survival rates meaningfully. Second, PTEN mutations are observed predominantly in endometrioid carcinomas and not in nonendometrioid subtypes, such as serous or clear cell, which are aggressive and have poor prognosis. [1][2][3]8 To examine the influence of PTEN, it is necessary to select endometrioid carcinoma.To clarify the prognostic significance of PTEN, we investigated the relationship between its expression and prognosis in a large number of patients with advanced endometrioid endometrial carcinoma. MATERIAL AND METHODS PatientsA total of 784 patients with endometrial carcinoma underwent primary treatment between 1985 and 2000 at Tottori University Hospital, Kurume University Hospital, National Defense Medical School Hospital, Jichi Medical School Hospital or Kawasaki Medical School Hospital (Japan). Of the 784 patients, 98 who met the following criteria were entered in the study: pure endometrioid carcinoma, undergoing surgical staging and positive retroperitoneal lymph nodes. All subjects underwent hysterectomy, bilateral salpingo-oophorectomy and pelvic and/or para-aortic lymphadenectomy. Postoperative treatment was performed according to each institutional protocol. As a result, 25 patients received wholepelvis and/or para-aortic irradiation and 62 patients received platinum-based chemotherapy. The remaining 11 patients did not have any treatment because of their poor physical condition. Eightyseven (88.8%) patients were in FIGO stage IIIc and 11 (11.2%) in stage IV. Mean age was 58.6 (range 30 -78) years. All patients provided informed consent for research use of their samples. Immunohistochemic stainingA 4 m section was cut from the paraffin block of a primary uterine tumor. Each section was mounted on a silane-coated glass slide, deparaffinized and soaked for 15 min at room temperature in 0.3% H 2 O 2 /methanol to block endogenous peroxidase. A mouse anti-PTEN monoclonal antibody, PTEN A2B1 (Santa Cruz Biotechnology, Santa Cruz, CA) was applied for ...

Chemosensitivity in cervical cancer may be associated with apoptosis via the p53-Bax pathway.