Self-Assembly Bifunctional Tetranuclear Ln2Ni2 Clusters: Magnetic Behaviors and Highly Efficient Conversion of CO2 under Mild Conditions.

A series of heterometallic tetranuclear clusters, Ln2Ni2(NO3)4L4(µ3-OCH3)2·2(CH3CN) (Ln = Gd(1), Tb(2), Dy(3), Ho(4), Er(5); HL = methyl 3-methoxysalicylate), were synthesized solvothermally. The intramolecular synergistic effect of two metal centers of Ln(III) and Ni(II) and the exposed multimetallic sites serving as Lewis acid activators greatly increase the efficiency of the CO2 conversion, and the yield for cluster 3 can be achieved at 96% at atmospheric pressure and low temperature. In particular, the self-assembly multimetal center with polydentate ligand shows good generality and enhanced recyclability. The design of such 3d-4f heterometallic clusters provides an effective strategy for the conversion of CO2 under greener conditions. Meanwhile, magnetic investigations indicate that cluster 1 is a good candidate for magnetic refrigerant materials with a relatively large magnetocaloric effect (MCE) (-ΔSm = 28.5 J kg-1 K-1 at 3.0 K and 7.0 T), and cluster 3 shows single-molecular magnet behavior under zero dc field. Heterometallic clusters with special magnetic properties and good catalytic behavior for the conversion of CO2 are rare. Thus, they are potential bifunctional materials applied in practice.

The Effects of miR-195-5p/MMP14 on Proliferation and Invasion of Cervical Carcinoma Cells Through TNF Signaling Pathway Based on Bioinformatics Analysis of Microarray Profiling.

BACKGROUND/AIMS: This study is aimed at identification of miR-195-5p/MMP14 expression in cervical cancer (CC) and their roles on cell proliferation and invasion profile of CC cells through TNF signaling pathway in CC. METHODS: Microarray analysis, gene set enrichment analysis (GSEA) and DAVID were used to analyze differentially expressed miRNAs, mRNAs and signaling pathways. MiR-195-5p and MMP14 expression levels in CC cell were determined by qRT-PCR. Western blot was employed to measure MMP14 and TNF signaling pathway-relating protein level. Luciferase reporter system was used to confirm the targeting relationship between MMP14 and miR-195-5p. Cell proliferation and invasion was respectively deeded by CCK8, transwell. In vivo experiment was carried out to study the impact of MMP14 and miR-195-5p on CC development in mice. RESULTS: The microarray analysis and the results of qRT-PCR determined that miR-195-5p was under-expressed and MMP14 was over-expressed in CC cells. GSEA and DAVID analysis showed that TNF signaling pathway was regulated by miR-195-5p/MMP14 and activated in cervical carcinoma cells. The miR-195-5p and MMP14 have a negative regulation relation. In vivo experiment found that down-regulated MMP14 and up-regulated miR-195-5p suppressed the tumor development. CONCLUSION: Our results suggest that MMP14 is a direct target of miR-195-5p, and down-regulated MMP14 and up-regulated miR-195-5p suppressed proliferation and invasion of CC cells by inhibiting TNF signaling pathway.

Elevated growth differentiation factor 15 expression predicts poor prognosis in epithelial ovarian cancer patients.

The purpose of this study was to determine the expression of growth differentiation factor 15 (GDF15) and explore its clinical significance in epithelial ovarian cancer (EOC) patients. The expression of GDF15 in EOC tissues and serum samples was evaluated using immunohistochemistry and enzyme-linked immunosorbent assay (ELISA), respectively. The association of GDF15 expression with clinicopathologic parameters was analyzed. Survival time was assessed using the Kaplan-Meier technique and Cox regression model. Both in EOC tissues and serum, high GDF15 levels were obviously related with advanced International Federation of Gynecology and Obstetrics (FIGO) stage, lymph node metastasis, ascites, and chemoresistance. Kaplan-Meier analysis indicated that EOC patients with high GDF15 expression showed poorer progression-free survival (PFS) and overall survival (OS). Multivariate analysis demonstrated that GDF15 expression was an independent predictor of PFS in EOC patients. Our study shows that elevated GDF15 expression was associated with poor prognosis in EOC patients. We suggest that GDF15 is a novel biomarker for the early detection of EOC, prediction of the response to chemotherapy, and screening for recurrence in EOC patients.

Platinum sensitivity and CD133 expression as risk and prognostic predictors of central nervous system metastases in patients with epithelial ovarian cancer.

BACKGROUND: To characterize prognostic and risk factors of central nervous system (CNS) metastases in patients with epithelial ovarian cancer (EOC). METHODS: A retrospective analysis of Xijing Hospital electronic medical records was conducted to identify patients with pathologically confirmed EOC and CNS metastases. In addition to patient demographics, tumor pathology, treatment regimens, and clinical outcomes, we compared putative cancer stem cell marker CD133 expression patterns in primary and metastatic lesions as well as in recurrent EOC with and without CNS metastases. RESULTS: Among 1366 patients with EOC, metastatic CNS lesions were present in 29 (2.1%) cases. CD133 expression in primary tumor was the only independent risk factor for CNS metastases; whilst the extent of surgical resection of primary EOC and platinum resistance were two independent factors significantly associated with time to CNS metastases. Absence of CD133 expression in primary tumors was significantly associated with high platinum sensitivity in both patient groups with and without CNS metastases. Platinum resistance and CD133 cluster formation in CNS metastases were associated with decreased survival, while multimodal therapy including stereotactic radiosurgery (SRS) for CNS metastases was associated with increased survival following the diagnosis of CNS metastases. CONCLUSIONS: These data suggest that there exist a positive association between CD133 expression in primary EOC, platinum resistance and the increased risk of CNS metastases, as well as a less favorable prognosis of EOC. The absence of CD133 clusters and use of multimodal therapy including SRS could improve the outcome of metastatic lesions. Further investigation is warranted to elucidate the true nature of the association between platinum sensitivity, CD133 expression, and the risk and prognosis of CNS metastases from EOC.

Quantitative assessment of the association between CYP1A1 A4889G polymorphism and endometrial cancer risk.

Cytochrome P450 1A1 (CYP1A1) A4889G polymorphism was supposed to be associated with endometrial cancer risk, but previous studies reported conflicting results. We therefore performed a meta-analysis of all relevant studies to get a comprehensive assessment of the association between CYP1A1 A4889G polymorphism and endometrial cancer risk. The pooled odds ratios (OR) with the corresponding 95% confidence interval (95% CI) was calculated to assess the association. Finally, ten studies with a total of 1,682 endometrial cancer cases and 2,510 controls were finally included into the meta-analysis. Meta-analysis of the total ten studies showed that CYP1A1 A4889G polymorphism was not associated with endometrial cancer risk (ORG versus A = 1.14, 95% CI 0.83-1.57, P OR = 0.417; ORGG versus AA = 1.23, 95% CI 0.70-2.18, P OR = 0.470; ORGG versus AA/AG = 1.03, 95% CI 0.59-1.81, P OR = 0.919; ORGG/AG versus AA = 1.22, 95% CI 0.82-1.81, P OR = 0.336). Subgroup analyses by ethnicity further showed that there was also no obvious association between CYP1A1 A4889G polymorphism and endometrial cancer risk in both Caucasians and Asians. Sensitivity analysis by excluding single study in turns showed that the pooled estimations were not stable. Therefore, evidence for currently available data suggests that CYP1A1 A4889G polymorphism is not associated with endometrial cancer risk. However, more studies with large number of participants are needed to further assess the association precisely.

Combination of fenretinide and selenite inhibits proliferation and induces apoptosis in ovarian cancer cells.

The combination of fenretinide and selenite on ovarian cancer cells was investigated to assess its effects on proliferation and ability to induce apoptosis. Our results showed that fenretinide and selenite in combination significantly suppress the proliferation of ovarian cancer cells and induced apoptosis (including reactive oxygen species generation, and the loss of mitochondrial membrane potential) compared with either drug used alone. The caspase3/9-dependent pathway was triggered significantly in combination treatment, and moreover, the AMPK pathway also mediated the apoptosis induction in fenretinide and selenite combination. Fenretinide and selenite combination treatment was demonstrated to suppress tumor growth in vivo, this drug combination has been thus found to have an enhanced anti-tumor effect on ovarian cancers cells.

[Multicenter randomized controlled clinical study for the operative treatment of malignant ovarian germ cell tumors].

OBJECTIVE: To investigate the operative treatment for first-treated patients with malignant ovarian germ cell tumors who need preservation of fertility. METHODS: The clinical data of 105 patients who were treated with fertility-sparing surgery in 11 hospitals from 1992 to 2010 were collected to evaluate the outcomes of different primary surgical operative procedures. All 105 cases were performed the surgeries that preserved fertility and divided into three groups according to the surgical approaches, comprehensive staging surgery group: 47 cases (44.8%) received comprehensive staging surgeries that including the ipsilateral oophorectomy + omentectomy + retropertoneal lymph node dissection ± appendectomy + multiple biopsies;oophorectomy group:45 cases (42.9%)received ipsilateral oophorectomy ± biopsy of contralateral ovary ± omentectomy;tumor resection group:13 cases (12.4%) received enucleation of the mass with preservation of the ovary. Differences were compared among the three groups of patients in the surgery-related indicators, complications, fertility and prognosis. RESULTS: (1) Surgery-related indicators:the average blood loss of the comprehensive staging surgery group, the oophorectomy group and the tumor resection group were 496, 104 and 253 ml, the mean operation time were 176, 114 and 122 minutes, respectively, and there were significant differences among three groups (P = 0.011, P = 0.000). (2) Complication:the surgical complication rates of the three groups were 17% (8/47), 0 and 1/13, with significant differences (P = 0.015). (3) Reproductive function status: the pregnancy rate and birth rate of the three groups were no significant differences (9/19 vs. 7/19 vs. 2/3, P = 0.515; 8/19 vs. 5/19 vs. 2/3, P = 0.636). (4) PROGNOSIS: the recurrence rate of the three groups were significant differences [13% (6/47) vs. 0 vs. 2/13, P = 0.013], but the death rate with no significant differences [6% (3/47) vs. 0 vs. 0, P = 0.129]; The five-year survival rate of three different groups were 89%, 100% and 100% (P > 0.05), while disease free survival rate were 85%, 100% and 83% (P < 0.05), respectively. CONCLUSIONS: Compared with comprehensive staging surgery, oophorectomy group have higher surgical security and satisfactory prognosis, considerable pregnancy rates and birth rate. The tumor resection security may be reliable, but the prognosis is poor.

Two novel Ln8 clusters bridged by CO32- effectively convert CO2 into oxazolidinones and cyclic carbonates.

It is difficult and challenging to design and construct high-nuclearity Ln(III)-based clusters due to the high coordination numbers and versatile coordination geometries of Ln(III) ions. Herein, two novel octanuclear Ln(III)-based clusters [Ln8(H2L-)4(HL2-)4(NO3)6 (CO3)2](NO3)2·2CH3CN (Ln = Nd (1) and Sm (2)) have been synthesized under solvothermal conditions. The X-ray single analysis reveals that both 1 and 2 are octanuclear structures and the eight central Ln(III) ions are bridged by two CO32- anions. Catalytic study revealed that 1 and 2 can effectively catalyze the cycloaddition reaction of CO2 and aziridines or epoxides simultaneously under mild conditions. What is more, cluster 1, as a heterogeneous catalyst, can be reused at least three times without obvious loss in catalytic activity for coupling of CO2 and epoxides. To our knowledge, cluster 1 is the first Ln(III)-based cluster catalyst used for the conversion of CO2 with aziridines or epoxides simultaneously. This work provides a successful strategy to integrate high-nuclear Ln(III)-based clusters for CO2 conversion, which may open a new space for the construction of multifunctional high-nuclear Ln(III)-based clusters as efficient catalysts for CO2 conversion.

Self-assembly of octanuclear Ln(III)-based clusters: their large magnetocaloric effects and highly efficient conversion of CO2.

The design and construction of high-nuclear lanthanide clusters with fascinating topology and functional properties have been an active area of research, however, the development of an effective approach for obtaining high-nuclear lanthanide clusters with multifunctional properties is still extremely difficult. Up to now, a systematic approach for guiding the further expansion of Ln(III)-based clusters showing good functional properties is lacking. Herein, we design and synthesize a polydentate Schiff base ligand (HL), which reacts with ß-diketonate salts Ln(acac)3·2H2O, and a series of Ln8 clusters [Ln8(acac)6(L)2(µ3-O)6(µ2-C2H5O)4(µ2-Hacac)2]·2CH3CN (Ln(III) = Gd (1), Dy (2), and Ho (3); HL = pyridine-2-carboxylic acid (5-hydroxymethyl-furan-2-ylmethylene)-hydrazide, Hacac = acetylacetone) have been successfully synthesized. Single-crystal X-ray diffraction studies reveal that clusters 1-3 are isostructural and can be viewed as a Ln8 core bridged by eighteen µ2-O atoms, six µ3-O atoms and two µ4-O atoms. Magnetic studies show that cluster 1-Gd8 displays a large magnetocaloric effect with -ΔSm = 46.14 J kg-1 K-1 (T = 2.0 K and ΔH = 7.0 T); cluster 2-Dy8 exhibits single-molecule magnet behavior under zero-field conditions. It is worth mentioning that the -ΔSm of cluster 1-Gd8 is larger than that of most reported polynuclear Gd(III)-based clusters; the 2-Dy8 cluster is one of the rare polynuclear Lnn SMMs (n ≥ 8) under zero dc field. Importantly, these Ln(III)-based clusters (1-3) can catalyze the cycloaddition of CO2 with epoxides with high efficiency under mild conditions; and cluster 1-Gd8 as a catalyst could be reused at least three times without obvious loss of catalytic performance.

GnRH antagonist cetrorelix inhibits mitochondria-dependent apoptosis triggered by chemotherapy in granulosa cells of rats.

OBJECTIVE: Exposure to chemotherapy causes various adverse effects on the ovaries including premature ovarian failure and infertility. GnRH antagonist cetrorelix could reverse the ovarian damage during chemotherapy, but the mechanism remains unclear. The objectives of this study were to examine the role of the cetrorelix for prevention of mitochondria-dependent apoptosis in granulosa cells of rats during the treatment with cyclophosphamide(Cy), if the mitochondria-dependent apoptotic process was involved. METHODS: Female SD rats were injected with cetrorelix before and after administration of saline, or Cy. Main outcome measures were the apoptotic indexes, serum hormones, ultrastructure of granulosa cells, mitochondrial membrane potential, the kinetics of cytochrome c (Cyt-c) processing in cells, and apoptotic markers. RESULTS: The ovarian apoptotic indexes as shown by TUNEL assay were reduced by cetrorelix pretreatment and the rats regained normal hormonal profile. The ultrastructure and JC-1 fluorescence intensity assessments showed cetrorelix pretreatment inhibited mitochondrial dysfunction in granulosa cells induced by chemotherapy. Western blot analysis showed that cetrorelix suppressed the release of Cyt-c from mitochondria to cytoplasm. Meanwhile, cetrorelix pretreatment expressed less Bax, caspase-3 and Cyt-c in granulosa cells compared with chemotherapy alone. CONCLUSION: Cetrorelix could reduce the apoptosis in granulosa cells through inhibiting mitochondria-dependent apoptosis triggered by chemotherapy.

The GnRH antagonist reduces chemotherapy-induced ovarian damage in rats by suppressing the apoptosis.

OBJECTIVE: GnRH antagonist cetrorelix could reserve the ovarian follicles during chemotherapy, but the mechanism remains unclear. The objectives of this study were to examine the overall effect of cetrorelix against ovarian failure and to define if the apoptotic process was involved. METHODS: Female SD rats were injected with cetrorelix before and after administration of saline, or cyclophosphamide (Cy), or oral etoposide (VP). Main outcome measures were the number of ovarian follicles, serum hormones, ovary histology and apoptotic markers. RESULTS: The females exposed to Cy or VP had reduced body and ovary weights, which could be restored by cetrorelix pretreatment. Single cetrorelix treatment could increase the number of primordial follicles, but reduce the number of growing and mature follicles. As a consequence, the ovaries exposed to cetrorelix prior to Cy or VP showed significantly higher numbers of follicles at all developing stages than those exposed to Cy or VP alone. Meanwhile, the ovarian apoptotic indexes as shown by TUNEL assay were reduced by cetrorelix pretreatment and the ovary expressed less caspases-3 and more Bcl-2 compared with chemotherapy alone. Moreover, the rats regained normal hormonal profile after cetrorelix pretreatment without any alterations in ovarian expression of estrogen receptor (ER)alpha, ERbeta, or progesterone receptor (PR). CONCLUSION: Cetrorelix could reduce the chemotherapy-induced ovarian damage through regulating the expression of Bcl-2 and caspases-3 in the ovary, without any expressional alterations of nuclear receptors, suggesting the apoptosis pathway involved.

Correlation between Aurora-A expression and the prognosis of cervical carcinoma patients.

OBJECTIVE: Aurora-A, a novel member of the serine/threonine kinase family, has been reported to be correlated with tumorigenesis. Our aim was to investigate whether Aurora-A expression correlates with clinicopathologic factors and prognosis of cervical carcinoma patients. DESIGN/SETTING: Retrospective study. POPULATION: Seventy-four cervical carcinoma patients, between 1996 and 2002. METHODS: Reverse transcription-polymerase chain reaction and Western blot assays were performed to detect the expression of Aurora-A gene in cervical carcinoma cells and paired cancerous and corresponding noncancerous tissues from 74 cervical carcinoma patients. The expression of Aurora-A protein in tissues was also determined by immunohistochemistry. The relationships of Aurora-A expression with clinical factors and prognosis of patients were evaluated by statistical analysis. RESULTS: The expression of Aurora-A mRNA and protein was significantly higher in cervical carcinoma cells than in normal cervical epithelial cell (p<0.05). The expression of Aurora-A mRNA in cancerous tissues was significantly higher than that in corresponding noncancerous tissues (p<0.001). The expression of Aurora-A protein was also increased in tumor tissues by immunochemistry. Aurora-A transcript expression was correlated with FIGO stage (p=0.018), tumor differentiation (p=0.014), parametrial invasion (p=0.024), lymphnode or hematogenous metastasis (p=0.005 or 0.019), but not other clinicopathological factors. Patients with high Aurora-A expression had a poorer disease-free survival and overall survival rates than patients with low Aurora-A expression. Multivariate analysis showed that high Aurora-A mRNA expression was an independent prognostic factor (risk ratio: 2.88; p=0.005). CONCLUSIONS: Aurora-A might be used as a prognostic marker for cervical carcinoma patients.

Inhibition of multi-drug resistance of ovarian carcinoma by small interfering RNA targeting to MRP2 gene.

AIMS: To investigate the inhibiting mechanism of multi-drug resistance (MDR) using expression vectors of short hairpin RNA (shRNA) in a MDR human ovarian cancer cell line (A2780/cp70). METHODS: Two shRNA expression vectors were constructed and introduced into A2780/cp70 cells. Expression of MRP2 mRNA was assessed by RT-PCR, and Mrp2 expression was determined by Western blot and immunocytochemistry. Apoptosis and sensitization of the cells to cisplatinum were quantified by flow cytometry and methyl-thiazol-tetrazolium (MTT) assays. Cellular cisplatinum accumulation was assayed by laser scanning confocal microscopy (LSCM). RESULTS: In A2780/cp70 cells transfected with MRP2-A and MRP2-B shRNA expression vectors, RT-PCR showed that MRP2 mRNA expression was reduced by 41.8% (P < 0.05), 30.9% (P < 0.01) (transient transfection) and 39.6% (P < 0.05), 29.4% (P < 0.01) (stable transfection), respectively. Western blot and immunocytochemistry showed that Mrp2 expression was significantly and specifically inhibited. Resistance against cisplatinum was decreased from 173- to 119-fold (P < 0.05), 64-fold (P < 0.01) (transient transfection) and to 117-fold (P < 0.05), 60-fold (P < 0.01) (stable transfection). Furthermore, shRNA vectors significantly enhanced the cellular cisplatinum accumulation. The combination of shRNA vectors and cisplatinum significantly induced the apoptosis of cells. CONCLUSIONS: shRNA expression vectors effectively reduce MRP2 expression and can restore the sensitivity of drug-resistant cancer cells to conventional chemotherapeutic agents.

NLRP3 inflammasome activation by estrogen promotes the progression of human endometrial cancer.

BACKGROUND: Activation of NLPR3 inflammasome is associated with the development and progression of some types of malignant tumors, but its role in endometrial cancer is unclear. This study aimed to investigate the expression and function of NLRP3 inflammasome in endometrial cancer. MATERIALS AND METHODS:  The expression levels of NLRP3, its inflammasome components and estrogen receptor ß in endometrial cancer and paired non-tumor tissues were detected. The effects of NLPR3 silencing or overexpression on the proliferation, migration, and invasion of Ishikawa and HEC-1A cells were determined. The impact of NLPR3 silencing on the growth of implanted tumors was determined in vivo. The effects of estrogen on NLPR3 inflammasome activation and Ishikawa cell proliferation were determined. RESULTS: The upregulation of NLRP3, ASC, caspase-1, and IL-1ß was associated with the progression of endometrial cancer and poor survival. NLPR3 silencing inhibited the proliferation, migration, and invasion of endometrial cancer cells while NLPR3 overexpression had opposite effects. NLPR3 silencing reduced IL-1ß and caspase-1 expression and the growth of implanted endometrial tumors, accompanied by decreased pro-IL-1ß maturation. Estrogen enhanced NLPR3, ERß, pro-IL-1ß, IL-1ß expression, and endometrial cancer cell proliferation, which were mitigated by treatment with ERß inhibitor but not ERα inhibitor. CONCLUSION: Our results suggest that estrogen acts through ERß to enhance the activation of NLPR3 inflammasome and promote the progression of endometrial cancer. NLPR3 inflammasome may be a new therapeutic target for endometrial cancer.

Survivin gene RNA interference inhibits proliferation, induces apoptosis, and enhances radiosensitivity in HeLa cells.

OBJECTIVE: Survivin is a new member of the inhibitors of apoptosis (IAPs) family. It is upregulated in various malignancies including human cervical carcinomas. Reduction of this molecule has resulted in chemosensitization, but it is uncertain whether it can lead to radiosensitization. We observed the effect of survivin gene RNA interference (RNAi) on the proliferation, apoptosis, and radiosensitivity of the human cervical carcinoma cells HeLa. STUDY DESIGN: Human cervical carcinoma cells (HeLa) were transfected with the specific siRNA expression vector (pSilencer2.1-s2) designed to target survivin mRNA. A corresponding site-mutated vector was constructed as a negative control (pSilencer2.1-NC). The expression of survivin mRNA and its protein among the stable transfected cells and the untransfected ones was detected by semi-quantitative RT-PCR and Western blotting respectively. The cell growth was examined by methyl thiazolyl tetrazolium (MTT) assay. The cell cycle distribution and cell apoptosis were measured by flow cytometry. The changes in cell radiosensitivity were observed by clonogenic survival assay. RESULTS: Three stable transfected cell lines: HeLa-s2 (with pSilencer2.1-s2), HeLa-NC (with pSilencer2.1-NC), and HeLa-U6 neo (with empty vector pSilencer2.1-U6 neo) were established. The expression levels of survivin gene mRNA and protein in HeLa-s2 were significantly lower than in HeLa-NC, HeLa-U6 neo, and those untransfected HeLa cells. The expression inhibitory rates were 62.8% and 60.1%. The cell proliferation of HeLa-s2 was inhibited, and the highest inhibitory rate was 57.8+/-2.1%. The changes in cell cycle distribution in HeLa-s2 compared with the other three cell lines were obvious, many cells were blocked in the G(0)/G(1) phase 72.7+/-3.1% (P<0.05), reduced sharply in the G(2)/M phase (5.1+/-2.9)% (P<0.05), and also the apoptotic rate was 29.2+/-1.4%, obviously increasing (P<0.05). At the same dose of radiation, the cloning efficiency of HeLa-s2 declined notably (P<0.05); the cell survival curve showed a significant decrease in D(0) and D(q), which were 3.15 and 1.21, respectively (P<0.05), and the radiation enhancement ratios were 2.01 (a ratio of D(0)) and 1.77 (a ratio of D(q)). CONCLUSIONS: Survivin gene RNAi not only could inhibit the proliferation of human cervical carcinoma cells (HeLa), but also could significantly enhance the radiosensitivity of those cells through the reduction of its mRNA and protein. Therefore, an RNAi-targeted survivin gene strategy would be a potential approach to radiosensitization therapy in human cervical carcinomas.

Inhibition of tumor angiogenesis by cell-permeable dominant negative SOX18 mutants.

Angiogenesis play a key roles in tumor growth, invasion and metastasis, and has become an attractive target for anticancer drug development. Though a number of anti-angiogenic agents had entered clinical trials, few of them could reproduce the spectacular results in cancer patients as that had been seen in pre-clinical tumor models. Therefore, exploring novel anti-angiogenic agents is highly deserved. SOX18, a member of the Sry-related HMG box-containing family of transcription factors, is expressed transiently in endothelial cells during the development of blood vessels. And mutations resulting in expression of dominant negative SOX18 have been shown to severely impair the vascular development. Recent research demonstrated that SOX18 is expressed during the initial steps of tumor vascularization and involved in regulation of the expression of the VEGF receptor Flk-1 and the vascular cell adhesion molecule-1 (VCAM-1). Moreover, allograft tumor growth in mice heterozygous for Ra(Op) (RaOp mice) which express a dominant negative mutant form of SOX18 (SOX18RaOp) that does not interact effectively with the endothelial partner protein MEF2C, was dramatically slower than that of wild-type mice. In this article, we postulate that recombinant cell-permeable dominant negative SOX18 mutants, prepared by fusion with protein transduction domains, would inhibit tumor angiogenesis with high efficiency by impairing endothelial tube formation. If the hypothesis was proved to be practical, the fusion proteins would show promise as single anti-angiogenic agents in cancer therapy.

[Inhibition of ovarian cancer growth by small interfering RNA targeting X-linked inhibitor of apoptosis gene].

OBJECTIVE: To construct an RNA interference vector to down-regulate X-linked inhibitor of apoptosis (XIAP) gene and study the RNA interference effect on the cell cycle and growth of ovarian cancer. METHODS: Oligonucleotides of 64 base pairs for hairpin RNA targeting XIAP were designed, chemically synthesized, annealed, and cloned into the pSUPER vector. After identification by restriction digestion, the correct vectors were transiently transfected into SKOV3 cells, a human ovarian cancer cell line. The XIAP mRNA was detected by RT-PCR. The proteins were detected by western blot and indirect immunofluorescence staining. Flow cytometry (FCM) analysis and methyl thiazolyl tetrazolium (MTT) assay method were applied to measure cell cycle, cell growth and sensitiveness to cisplatin. RESULTS: SKOV3 cells had a high level expression of XIAP. The vector of RNA interference, which can interfere with XIAP gene was successfully constructed. After transient transfection, the expression of XIAP protein was significantly decreased in SKOV3 cells and the value of relative density was 3584 +/- 124, 2138 +/- 65, 1973 +/- 80 and 110 +/- 12, respectively (P = 0.0334). At the same time, the expression of XIAP mRNA was decreased accordingly and the value of relative density was 6674 +/- 274, 4532 +/- 107, 2322 +/- 57 and 1864 +/- 78, respectively (P = 0.0127). The FCM results showed that, the vector could increase the number of cells in G(1) phase compared with parent cells and compared with the cells transfected with pSUPER (P < 0.05); the number in S phase decreased compared with parent cells and compared with the cells transfected with pSUPER (P < 0.05). MTT results showed that pSUPER-siXIAP could inhibit the growth and proliferation of SKOV3 cells in a time-dependent manner, and inhibit the tumor cell growth. CONCLUSIONS: We successfully constructed an expressing hairpin RNA against the XIAP vector. The vector can effectively silence XIAP gene, increase the number of cells in G(1) phase, and slow down the growth of tumor cells. This may be a useful therapeutic strategy for XIAP over-expressing ovarian carcinomas.

Transcriptomic analysis of stromal cells from patients with endometrial carcinoma.

Tumor microenvironment plays a critical role in cancer pathogenesis. In this study, we performed transcriptomic analysis of stromal cells from patients diagnosed with endometrial carcinoma. Endometrial stromal cells from patients and healthy donors were cultured and total RNA was extracted for RNA integrity examination and gene profiling analysis. Gene ontology (GO) and KEGG analysis were also performed. In this study, we found that, in endometrial stromal cells from endometrial cancer patients, a total of 605 genes were changed (fold change ≥2, p-value <0.05). From these, 275 were up-regulated and 330 were down-regulated genes. In addition, GO analysis showed that the differentially expressed genes (DEGs) were involved in various biological processes including cell adhesion, biological adhesion, bone development and extracellular matrix organization. Furthermore, KEGG analysis of the DEGs identified four pathways including Wnt signaling pathway, cadherin signaling pathway, ECM-receptor interaction, and focal adhesion. Our study identified 605 DEGs in stromal cells from endometrial carcinoma which mapped to a variety of biological processes. These results may contribute to understanding the molecular mechanisms o of endometrial carcinoma pathogenesis.

Knockdown of RAB25 expression by RNAi inhibits growth of human epithelial ovarian cancer cells in vitro and in vivo.

AIMS: Ovarian cancer is the leading cause of cancer death among gynaecological malignancies. Elevated expression of Rab25 has been seen in this malignancy. To better understand its role in maintaining the malignant phenotype, we used RNA interference (RNAi) directed against Rab25 in our study. RNAi provides a new, reliable method to investigate gene function and has the potential for gene therapy. The aim of the study was to examine the anti-tumour effects elicited by a decrease in the level of Rab25 by RNAi and its possible mechanism of action. METHODS: According to the Rab25 mRNA sequence in Genbank, a pair of 64 nt oligonucleotides were designed and synthesised, each containing the sites of restriction endonuclease at both ends. Oligonucleotides were annealed and ligated with linearised pSUPER by ligase. The recombinants (named pSUPER/Rab25 siRNA) were finally sequenced and identified by enzyme cutting and sequencing. The human ovarian cell line A2780 was grown without transfection, transfection with empty vector and with pSUPER/Rab25 siRNA with electroporation. The inhibitory effect was examined by RT-PCR, MTT, FCM and tumour growth of athymic nude mice. RESULTS: Rab25 siRNA expression vector was successfully constructed and identified by double endonuclease digestion. Sequence analysis of inserted fragment revealed the same sequence as synthesised siRNA oligonucleotides. Cells transfected with Rab25 siRNA can specifically knock down the transcription of Rab25, exhibiting cells with slower proliferation, increased apoptosis, and decreased tumour growth. CONCLUSIONS: Rab25 siRNA expression vector has been successfully constructed, and it could inhibit the tumour growth both in vitro and in vivo. Our data suggest that the Rab25 signalling pathway plays a role in the regulation of cell proliferation and apoptosis in ovarian cancer cells, which indicates that the Rab25 gene plays a definite role in the development and aggressiveness of human ovarian cancer and should be further elucidated as a possible therapeutic target of ovarian cancer.