KDM4B and KDM4A promote endometrial cancer progression by regulating androgen receptor, c-myc, and p27kip1.

Epidemiological evidence suggests that elevated androgen levels and genetic variation related to the androgen receptor (AR) increase the risk of endometrial cancer (EC). However, the role of AR in EC is poorly understood. We report that two members of the histone demethylase KDM4 family act as major regulators of AR transcriptional activityin EC. In the MFE-296 cell line, KDM4B and AR upregulate c-myc expression, while in AN3CA cells KDM4A and AR downregulate p27kip1. Additionally, KDM4B expression is positively correlated with AR expression in EC cell lines with high baseline AR expression, while KDM4A and AR expression are positively correlated in low-AR cell lines. In clinical specimens, both KDM4B and KDM4A expression are significantly higher in EC tissues than that in normal endometrium. Finally, patients with alterations in AR, KDM4B, KDM4A, and c-myc have poor overall and disease-free survival rates. Together, these findings demonstrate that KDM4B and KDM4A promote EC progression by regulating AR activity.

Tyrosine kinase inhibitors induce mesenchymal stem cell-mediated resistance in BCR-ABL+ acute lymphoblastic leukemia.

Tyrosine kinase inhibitors (TKIs) are used as a frontline therapy for BCR-ABL(+) acute lymphoblastic leukemia (ALL). However, resistance to TKI therapy arises rapidly, and its underlying molecular mechanisms are poorly understood. In this study, we identified a novel cascade of events initiated by TKIs and traversing through mesenchymal stem cells (MSCs) to leukemic cells, leading to resistance. MSCs exposed to TKIs acquired a new functional status with the expression of genes encoding for chemo-attractants, adhesion molecules, and prosurvival growth factors, and this priming enabled leukemic cells to form clusters underneath the MSCs. This cluster formation was associated with the protection of ALL cells from therapy as leukemic cells switched from BCR-ABL signaling to IL-7R/Janus kinase signaling to survive in the MSC milieu. Our findings illustrate a novel perspective in the evolution of TKI resistance and provide insights for advancing the treatment of BCR-ABL(+) ALL.

Tumor heterogeneity in the recurrence of epithelial ovarian cancer demonstrated by polycomb group proteins.

PURPOSE: To investigate tumor heterogeneity in the recurrence of epithelial ovarian cancer demonstrated by polycomb group (PcG) proteins. METHODS: Tissue microarrays containing matched primary and recurrent ovarian tumors from the same patients were constructed for detection of PcG protein expression. Survival analyses of clinicopathological parameters and expression of PcG proteins were performed on progression-free survival (PFS) and overall survival (OS) of patients. Genetic and epigenetic heterogeneity was explored in aspects of gene copy number and microRNA (miRNA) profiling. RESULTS: PcG proteins were heterogeneously expressed in primary versus recurrent tumors (P<0.05). In univariate survival analysis of the ovarian carcinoma cohorts, a significant association of intensive expression of BMI1 and EZH2 in first-onset lymph node metastases with shortened PFS was demonstrated (P=0.010, P=0.019); and a significant association of intensive expression of BMI1 and EZH2 in recurrent tumors with shortened OS was demonstrated (P=0.042, P=0.047). Importantly, BMI1 and EZH2 expression provided significant independent prognostic parameters in multivariate analyses (P<0.05). Gene amplification did not always coincide with PcG protein expression. Eight miRNAs were found to be downregulated in recurrent tumors, among which miR-298 might indirectly regulate the expression of EZH2 through transcription factor ILF3. CONCLUSION: Tumor heterogeneity exists in the recurrence of epithelial ovarian cancer, manifested by PcG protein expression and underlying genetic and epigenetic alterations. Intensive expression of BMI1 and EZH2 are predictors of earlier relapse and shorter OS, independent of grade and chemotherapy sensitivity. EZH2 and miR-298 have great potential to be new targets for treatment of recurrent ovarian cancer.

PI3K/AKT/mTOR signaling pathway as a therapeutic target for ovarian cancer.

BACKGROUND: Ovarian cancer is one of the major causes of death in women worldwide. Despite improvements in conventional treatment approaches, such as surgery and chemotherapy, a majority of patients with advanced ovarian cancer experience relapse and eventually succumb to the disease; the outcome of patients remains poor. Hence, new therapeutic strategies are urgently required. The phosphatidylinositol 3-kinase (PI3K)/AKT/mammalian target of rapamycin (mTOR) is activated in approximately 70 % of ovarian cancers, resulting in hyperactive signaling cascades that relate to cellular growth, proliferation, survival, metabolism, and angiogenesis. Consistent with this, a number of clinical studies are focusing on PI3K pathway as an attractive target in the treatment of ovarian cancer. In this review, we present an overview of PI3K pathway as well as its pathological aberrations reported in ovarian cancer. We also discuss inhibitors of PI3K pathway that are currently under clinical investigations and the challenges these inhibitors face in future clinical utility. METHODS: PubMed was searched for articles of relevance to ovarian cancer and the PI3K pathway. In addition, the ClinicalTrials.gov was also scanned for data on novel therapeutic inhibitors targeting the PI3K pathway. RESULTS: Genetic aberrations at different levels of PI3K pathway are frequently observed in ovarian cancer, resulting in hyperactivation of this pathway. The alterations of this pathway make the PI3K pathway an attractive therapeutic target in ovarian cancer. Currently, several inhibitors of PI3K pathway, such as PI3K/AKT inhibitors, rapamycin analogs for mTOR inhibition, and dual PI3K/mTOR inhibitors are in clinical testing in patients with ovarian cancer. CONCLUSIONS: PI3K pathway inhibitors have shown great promise in the treatment of ovarian cancer. However, further researches on selection patients that respond to PI3K inhibitors and exploration of effective combinatorial therapies are required to improve the management of ovarian cancer.

Molecular portraits of heterogeneity related to cancer stem cells in human ovarian cancer.

PURPOSE: The aim of this study was to investigate molecular portraits of heterogeneity related to cancer stem cells (CSCs) in human ovarian cancer and to access the value in diagnosis and treatment. METHODS: Sixty specimens were collected in both cytoreductive and re-cytoreductive surgeries of 20 serous papillary ovarian adenocarcinoma cases. Expression density and distribution of 3 CSC markers (CD44, CD133, and CD117) and 3 stemness proteins (Bmi1, Nestin, and Oct3/4) were analyzed by immunohistochemical staining. Pairwise comparisons were performed among their expression in primary, metastasis, and relapsing tumors. RESULTS: Some molecules presented different localization in 1 tissue, like CD133 and CD117, and all but Oct3/4 expressed differentially in different specimens of 1 case. Compared to primary or metastatic cancers, recurrent cancers show higher expression of CD133, CD117, and Bmi1, as well as higher histological grades. CONCLUSIONS: Our study indicated that there exist extratumoral and intratumoral heterogeneity in ovarian epithelial cancers related to CSCs. And this is worth further studying.

Identification of a cancer stem cell-like side population in the HeLa human cervical carcinoma cell line.

The present study aimed to identify the stem cell characteristics of side population (SP) cells sorted from the widely-used HeLa human cervical carcinoma cell line. The SP cells were sorted from the HeLa cell line using fluorescence-activating cell sorting (FACS). Stem cell characteristics of the SP cells, including proliferation, self-renewal, differentiation and the ability to form xenografts, were investigated in vitro and in vivo. The SP cells demonstrated strong tumorigenesis following in vivo transplantation into five to six-week-old female Balb/c mice. The SP cells were observed to be more resistant to chemotherapy and radiotherapy compared with non-side population (NSP) cells. A higher expression of CD133 was observed in the SP cells compared with the NSP cells following FACS. The results demonstrated that the SP cells from the HeLa human cervical carcinoma cell line exhibit stem cell characteristics in vitro and also have a strong ability to form tumors in vivo. The cell surface marker CD133 may serve as a potential molecular marker for the identification of cervical cancer stem cells (CSCs).

[Retrospective investigation on changes of endemic situation before and after reaching criteria of schistosomiasis transmission controlled or interrupted in hilly endemic areas of Jiangxi Province].

OBJECTIVE: To investigate the changing rules of schistosomiasis endemic situation before and after reaching the criteria of schistosomiasis transmission controlled or interrupted in hilly endemic areas of Jiangxi Province, so as to provide the evidence for reformulating the criteria of schistosomiasis control and eradication in the future. METHODS: In the hilly areas of schistosomiasis endemic in Jiangxi Province, 2 counties where the transmission has been interrupted and 1 county where the transmission has been controlled were selected and investigated with the retrospective research method. The endemic detailed data were collected and recorded 10 years before reaching the criteria of transmission interrupted/controlled, and several years after reaching the criteria (ending in 2008), and then a database was established. The changing rules of endemic situation before and after reaching the criteria of transmission interrupted/controlled were analyzed and compared. RESULTS: After reaching the criteria of transmission controlled, in the 3 counties, Guangfen, Shangrao and Dean, the declined rates of areas with Oncomelania hupensis snails were 96.79%, 98.99%, and 99.77% respectively. The snail density maintained a lower level, and 95% of infected persons and cattle were cured. The average time from transmission controlled to the transmission interrupted was 17 years in Guangfen County and 26 years in Dean County. However, in Shangrao County, the snail situation rebounded due to the snail re-found and spread although the schistosomiasis morbidity of population/animals maintained stably. CONCLUSIONS: After reaching the criteria of transmission interrupted/controlled, the remained snails were easy to re-find and spread under some certain condition, which is one of main obstacles for reaching the criteria of transmission interrupted. In an isolated snail unit, if the snail area and snail density are controlled in a very low level, it is still difficult to transmit and spread schistosomiasis even if there exist infectious sources.

[Analysis of the characteristics of side population cells in the human ovarian cancer cell line OVCAR-3].

OBJECTIVE: To identify the presence of side population (SP) cells in human ovarian cancer cell line OVCAR-3 and to investigate whether SP cells have the characteristics of cancer stem cells. METHODS: SP and non-SP (NSP) cells from OVCAR-3 were isolated by fluorescence-activated cell sorting after being stained by DNA-binding dye Hoechst 33342. Limiting dilution transplantation assay, real-time PCR, and drug sensitivity assay were performed to compare the tumorigenic ability, differentiation ability in vivo, the mRNA expression of "stemness" marker (Oct-4, Klf4, and Nanog) and ATP-binding cassette (ABC) transporter (ABCG2, ABCB1, and ABCC2), and response to multiple drugs (cisplatin, paclitaxel, doxorubicin, and mitoxantrone) between SP and NSP cells. RESULTS: A few of SP cells [(1.13 ± 0.39)%] which were sensitive to reserpine were identified in OVCAR-3 cells. The injection of as few as 10(2) SP cells initiated tumors in two of five mice. Tumor latency was 52 - 61 days. However, the NSP cells did not generate any tumors in mice until 10(4) NSP cells were injected (two of five mice). Tumor latency was 64 - 98 days. Tumorigenicity of SP cells was enhanced by at least 100-fold than that of NSP cells. The SP cells regenerated both SP [(2.09 ± 0.73)%] and NSP populations in vivo with a fraction size that was comparable to the original population. The mRNA expression of "stemness" genes Oct-4, Klf4 and ABC transporters ABCG2, ABCC2 genes were elevated in SP cells compared to NSP cells, the fold changes were 1.95 ± 0.41 (P < 0.05), 4.26 ± 0.63 (P < 0.01), 3.22 ± 0.36 (P < 0.01), and 1.76 ± 0.26 (P < 0.01), respectively. The relative activity of SP and NSP cells were 0.757 ± 0.105 versus 0.474 ± 0.035 (P < 0.01), 0.521 ± 0.092 versus 0.384 ± 0.073 (P < 0.05), 0.742 ± 0.051 versus 0.526 ± 0.088 (P < 0.01), and 0.690 ± 0.096 versus 0.466 ± 0.112 (P < 0.01) when they exposed to 0.25 µg/ml cisplatin, 0.01 µmol/L paclitaxel, 0.25 µmol/L doxorubicin, and 0.05 µg/ml mitoxantrone, respectively. CONCLUSIONS: SP cells from OVCAR-3 have enhanced self-renewal, differentiation, and tumor-initiating capacity compared to NSP cells. The mRNA expression of stemness genes and ABC transporters are markedly elevated in SP cells, which showed resistance to multiple chemotherapeutic drugs and have characteristics of cancer stem-like cells. Therefore, SP phenotype could be used as a marker to isolate the cancer stem-like cells in ovarian cancer.

Ovarian cancer cells with the CD117 phenotype are highly tumorigenic and are related to chemotherapy outcome.

Cancer stem cells (CSCs) play an important role in the recurrence and drug resistance of cancer. Isolation and characterization of CSCs from ovarian cancer samples may help to provide novel diagnostic and therapeutic targets in the management of recurrent disease and drug resistance in ovarian cancer. Here, we developed a xenograft model in which cells from 14 samples of human ovarian serous adenocarcinoma tissue or ascites were implanted in immunodeficient mice to test the tumorigenic potential of different populations of ovarian cancer cells. We identified and isolated the tumorigenic cells as CD117(+)Lineage(-) from three different xenografts. As few as 10(3) cells with the CD117(+)Lineage(-) phenotype, which comprise <2% of the xenograft tumor cells, were able to regenerate tumors in a mouse model, a 100-fold increase in tumorigenic potential compared to CD117(-)Lineage(-) cells. The tumors that arose from purified CD117(+)Lineage(-) cells reproduced the original tumor heterogeneity and could be serially generated, demonstrating the ability to self-renew and to differentiate, two defining properties of stem cells. Furthermore, immunohistochemistry analysis of 25 patients with advanced ovarian serous adenocarcinoma revealed positive immunostaining for CD117 in 40% (10 of 25) of patients. CD117 expression was statistically correlated with resistance to conventional chemotherapy (P=0.027). In conclusion, our study demonstrates that human ovarian cancer cells with the CD117(+) phenotype possess the unique properties of CSCs, including self-renewal, differentiation, a high tumorigenic potential, and chemoresistance. Future studies designed to target CD117(+) cancer cells may identify more attractive and effective therapies for treatment of ovarian cancer.

[Differential expression of microRNAs associated with estrogen receptor alpha and progesterone receptor in typeIand typeII endometrial adenocarcinomas.].

OBJECTIVE: To identify differentially expression of microRNAs associated with expression of estrogen receptor alpha (ERalpha) and progesterone receptor (PR) between type I and type II endometrial adenocarcinoma. METHODS: Two kinds of endometrial adenocarcinoma cell lines, Ishikawa and KLE, was transplanted into nude mice and biopsied to identify the expression of ERalpha, PR and p53, and test their response to estrogen and progesterone. Cultured the two cell lines under the estrogen-free and progesterone-free circumstance, total RNA was isolated to identify the differentially expressed microRNAs by microarray for prediction the microRNAs which target ESR1 and PGR by software miRANDA and TargetScan, and then was validated by real-time PCR in two cell lines cultured both in vivo and in vitro and ten specimens from patients. RESULTS: Ishikawa cell line was confirmed from type I endometrial adenocarcinoma, KLE cell line was confirmed from type II endometrial adenocarcinoma. One hundred and twenty-six differentially expressed microRNAs between the two cell lines were identified by microRNA microarray, among of which may target ESR1 included hsa-miR-100, 99a, and may target PGR included hsa-miR-378, 768-3p. The differential expression of hsa-miR-100, 99a, 378, 768-3p identified by microarray between Ishikawa and KLE in vivo and in vitro was equal to that by real-time PCR, while Hsa-miR-100 was significantly down expressed in type I group specimens compared to type II group (P < 0.01). CONCLUSION: Hsa-miR-100 is significantly down-expressed in type I endometrial adenocarcinoma compared to type II, which may be a great potential to target ESR1.

[Expression of MIF, VEGF and p16 proteins and their correlation with clinicopathological features in cervical cancer].

OBJECTIVE: To investigate the expression of macrophage migration inhibitory factor (MIF), p16 and vascular endothclial growth factor (VEGF) proteins and their relationship with clinicopathological features in cervical cancer. METHODS: Tissue microarray (TMA) and immunohistochemistry were used to detect the expression of MIF, p16 and VEGF proteins in specimens of 10 normal cervical epithelial tissues, 18 cervical intraepithelial neoplasia (CIN II, III) and 31 cervical squamous cell carcinomas. Western blotting was used to detect the expression of MIF, p16 and VEGF proteins in fresh samples of 3 normal cervical epithelial tissues, 3 CIN (III) and 6 cervical squamous cell carcinomas (3 Ib and 3 IIb). RESULTS: Positive expression rates of MIF were 0, 72.2% and 93.5% in the normal, CIN and carcinoma samples, 20.0%, 33.3% and 71.0% for p16, and 10.0%, 44.4% and 74.2% for VEGF, respectively. The expression rates and levels of the three genes were significantly higher in cervical carcinomas than those in CIN. MIF expression was significantly higher in the cases with lower differentiation (17 cases, P = 0.021), and was positively correlated with VEGF expression (P = 0.0045). VEGF expression rate was significantly higher in both cases of poorly differentiated carcinomas and those with stage II b carcinoma or beyond (P = 0.004, P = 0.008). p16 expression was not found to be correlated with tumor differentiation or clinical stage. It was showed by Western blotting that the expression levels of MIF, VEGF and p16 were significantly higher in the carcinomas than those in CIN or normal tissues. CONCLUSION: Expression of MIF, VEGF and p16 are probably involved in the process of cervical carcinogenesis. MIF expression is correlated with tumor differentiation. VEGF expression is correlated with both tumor differentiation and clinical stage.