Stellettin B, isolated by wild-type sponge Stelletta sp. from the ocean, exhibited potent antiprolifertive activities on various tumor cells. In this study, we use demonstrated the stellettin B inhibited the migration and invasion of the hepatocellular carcinoma cell HA22T/VGH. We found that the stellettin B inhibited the invasion and migration of hepatocellular carcinoma cells in a dose-dependent manner. The results of zymography assay showed that stellettin B suppressed the activities of matrix metalloproteinase MMP-9 and MMP-2. Moreover, protein levels of MMP-9, MMP-2, and urokinase-type plasminogen activator (uPA) were reduced by stellettinB in a dose-dependent manner. Stellettin B also exerted an inhibitory effect on phosphorylation of c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinases (ERK), phosphatidylinositol 3-kinase (PI3K) and Akt. Taken together, these results demonstrated that stellettin B could inhibit hepatocellular carcinoma cell migration and invasion and alter HA22T/VGH cell metastasis by reduction of uPA, MMP-2 and MMP-9 expression through the suppression of MAPKs and FAK/PI3K/AKT/mTOR signaling pathway. These findings suggest that stellettin B merits further evaluation as a chemotherapeutic agent for human hepatocellular carcinoma.

Three new cembranoids, lobophylides A-C (1-3), along with three known compounds (4-6) were isolated from the Formosan soft coral Lobophytum sp. The structures of these compounds were elucidated by means of IR, MS and NMR techniques and comparison of the NMR data with those of known analogues. Among these natural products, compound 2 is rarely found in cembrane diterpenes possessing an isopropyl moiety with an epoxide group. Cytotoxicity study of 1-6 against the growth of K562, Molt-4 and HL-60 cancer cell lines showed that compounds 1-6 were not cytotoxic (IC50 > 20 µg/mL) toward the above three cancer cell lines. The in vitro anti-inflammatory effects of compounds 1-5 were also tested. The results showed that compounds 2 and 3 significantly suppressed the accumulation of pro-inflammatory proteins, iNOS and COX-2.

Bioactivity-guided fractionation of the marine sponge Aaptos sp. extract led to the isolation of aaptamine (1), demethyloxyaaptamine (2) and isoaaptamine (3). The cytotoxic activity of the isolated compounds (1-3) was evaluated revealing that compound 3 was the most potent cytotoxic against breast cancer T47D cells. In a dose dependent manner, 3 inhibited the growth of T47D cells as indicated by the short- (MTT) and long-term (colony formation) assays. The cytotoxic effect of 3 was mediated through apoptosis which was suggested by DNA ladder formation, caspase-7 activation, XIAP inhibition and PARP cleavage. Furthermore, TEM and flow cytometric analysis using acridine orange dye indicated that that 3 treatment could induce t-47D cells autophagy. Immunoblot assays demonstrated that 3 treatment significantly activated autophagy marker proteins such as the increase in Type II LC-3. In addition, 3 treatment enhanced the activation of DNA damage (γH2AX) and ER stress-related proteins (IRE1αand BiP). Moreover, the use of 3 resulted in a significant increase in the generation of reactive oxygen species (ROS) as well as in the disruption of mitochondrial membrane potential (MMP). The pretreatment of t-47D cells with an ROS scavenger, NAC, attenuated apoptosis- and MMP disruption-induced by 3 up to 90%.Taken together, these findings suggest that the anticancer effect of 3 is associated with the induction of apoptosis and autophagy through oxidative stress. Accordingly, our data indicated that 3 exhibited as an outstanding lead for the development of marine derived anti-breast cancer agent.

One new 10-demethylated steroid, nephtheasteroid A (1), one new 19-oxygenated steroid, nephtheasteroid B (2), and four known steroids (3-6) were isolated from the wild-type soft coral Nephthea erecta. Two new cembranoids, culobophylins D (7) and E (8), along with eight known compounds (9 -16) were isolated from the cultured soft coral Lobophytum crassum. The structures were elucidated by means of IR, MS, and NMR techniques, and comparison of the NMR data with those of known analogues. Compound 1 was found to have a novel skeleton with the C10 demethylation of the normal steroid. Evaluation of the cytotoxicities showed that compound 9, the most potent of compounds 1-16, exhibited cytotoxicity against the K562, Molt-4, U937, Sup-T1 and Ca9 – 22 cancer cell lines with IC50 values of 1.2, 0.4, 2.4, 0.5 and 1.1 µg/mL, respectively. Moreover, compound 10 also showed significant U937 and Sup-T1 inhibitory activity, with IC50 values of 1.87 and 1.13 µg/mL.

Marine sponge of the genus Luffariella was found to be a rich source of novel cytotoxic and anti-inflammatory sesterterpenoids from previous reports [1, 2]. The further chemical investigation on this sponge led to the discovery of five new sesterterpenoids, luffarlides G-K, which were elucidated by spectroscopic methods including 1D and 2D NMR techniques. The absolute configurations were determined utilizing ECD/CD experiments. The antileukemic activity of the new compounds, along with the six previously described sesterterpenoids, were evaluated against Molt4 human acute lymphoblastic leukemic cell lines using MTT proliferative assay [3]. The most active compound, manoalide, was found to exhibit strong anti-proliferative activitie with IC50 values of 0.14 µg/mL. In addition, manoalide treatment activated caspase-related apoptotic proteins and mitogen-activated protein kinases (MAPK) of Molt4 cells with Western blot assay [3]. Analyzed by flow cytometry, manoalide treatment induced cancer cells apoptosis via disruption of mitochondrial membrane potential and generation of intracellular reactive oxygen species (ROS) in Molt4 cells. Manoalide could inhibit both activities of human topoisomerase I and II with gel electrophoresis [3]. DNA damage responses were directly detected with increase of H2AX phosphorylation, biomarker of DNA damage, and DNA break by comet assay. Importantly, the apoptosis-caused by manoalide could be recovered with NAC pretreatment, but not with pretreatments of caspase and MAPK inhibitors. Taken together, these results suggested that cytotoxic effect of manoalide is to mediate ROS generation, and finally resulted in cellular DNA damage and apoptosis. The findings provide support for further ChemGPS-NP targets investigation as well as the in vivo evaluation of lead compound.

Five new cembranoids, sandenlides A (1) and B (2), 3-epi-diepoxycembrene A (6), and flexibinoids A (8) and B (9), along with eleven known related metabolites 3-5, 7 and 10-16 have been isolated from the two cultured soft corals Sinularia sandensis and Sinularia flexibilis. The structures were elucidated by means of IR, MS, and NMR techniques, and the absolute configurations of 1, 4, 9 and 15 were further confirmed in conjunction with a single-crystal X-ray diffraction analysis. In the in vitro anti-inflammatory effects test, compounds 9-14 were found to significantly inhibit the accumulation of the pro-inflammatory iNOS and COX-2 proteins of the LPS-stimulated RAW264.7 macrophage cells. Structure-activity relationships analysis indicated that cembrane-type with one seven-membered lactone moiety at C-1 are potential anti-inflammatory agents. Furthermore, to the best of our knowledge, this is the first farming system for Sinularia sandensis in the world.

The mammalian Janus kinase (JAK) family consists of four members, namely JAK1, JAK2, JAK3 and TYK2, which play a critical role in cytokine/growth factor signaling and is increasingly associated with human cancers. Aberrant activation of these non-receptor tyrosine kinases may contribute to carcinogenesis. Herein, we focused on exploring the potential role of p-JAK1 in breast cancer. The expression profiles of p-JAK1 were analyzed in 68 pairs of cancer and non-cancer breast tissues from the same infiltrating ductal carcinoma case by using immunoblotting technique. The results obtained were further correlated with clinicopathological characteristics. Intriguingly, p-JAK1 expression was decreased in 55.9% of breast cancer tissues as compared to the matched non-cancer tissues. Further immunohistochemistry study showed an intense p-JAK1 staining predominantly in adjacent normal breast tissues but not the matched cancer lesions. Decreased p-JAK1 expression in breast cancer tissues was significantly correlated with positive estrogen receptor (ER) status and increased tumor size (p=0.010 and 0.009). We also found that p-JAK1 expression was high in ERalpha-negative breast cancer cell lines but was low in ERalpha-positive breast cell lines. Transfection of ERalpha-positive MCF-7 cells with an ERalpha-specific siRNA upregulated the expression of p-JAK1. In summary, our results indicated that an altered p-JAK1 expression might be involved in the development of breast infiltrating ductal carcinoma in an ERalpha-related manner.

Objective. To analyze whether leptin levels of the amniotic fluid elevate during early pregnancy in women destined to develop preeclampsia and to evaluate the relationship between amniotic fluid leptin levels and gestational age, maternal body mass index, and fetal sex. Study design. Leptin levels of the amniotic fluid were compared in two groups of women, preeclamptic ( n = 20) and normotensive pregnant ( n = 40), matched for fetal sex, maternal body mass index at sampling, gravidity and fetal gestational age at sampling. Furthermore, amniotic leptin levels in 400 normotensive pregnant women were analyzed for their correlation with gestational age, maternal body mass index, and fetal sex. Results. Median leptin concentrations were significantly higher ( p< 0.001) in the women with preeclampsia (7.3 +/- 0.7 ng/ml) than in the normotensive pregnant women ( 4.1 +/- 0.3 ng/ml), independent of fetal sex. The leptin levels in the amniotic fluid decreased with advanced gestational age ( r = 0.24, p< 0.001). Amniotic fluid leptin levels in the pregnant women carrying a female fetus (5.6 +/- 0.3 ng/ml) were significantly higher than those carrying a male fetus (4.7 +/- 0.2 ng/ml) ( p = 0.004). Conclusion. Higher amniotic fluid leptin levels were observed in the preeclamptic pregnant women, and they decreased as gestational age advanced. Furthermore, the women with a female fetus were noted to have higher amniotic fluid leptin levels.

Annonaceous acetogenins are a group of potential anti-neoplastic agents isolated from Annonaceae plants. We purified squamocin , a cytotoxic bis-tetrahydrofuran acetogenin, from the seeds of Annona reticulata and analyzed its biologic effects on cancer cells. We showed that squamocin was cytotoxic to all the cancer lines tested. Furthermore, squamocin arrested T24 bladder cancer cells at the G1 phase and caused a selective cytotoxicity on S-phase-enriched T24 cells. It induced the expression of Bax and Bad pro-apoptotic genes, enhanced caspase-3 activity, cleaved the functional protein of PARP and caused cell apoptosis. These results suggest that squamocin is a potentially promising anticancer compound.

Objectives. To explore the expression patterns and possible involvement of leptin and its receptor in the pathogenesis of urinary bladder cancer, with a focus on transitional cell carcinoma.Methods. Using reverse transcription-polymerase chain reaction, immunoblotting, and immunohistochemistry techniques, we correlated the expression patterns of leptin and its receptor with the occurrence of transitional cell carcinoma. We also applied transient transfection followed by BrdU labeling and immunofluorescent staining to address the effect of the leptin receptor on bladder cancer cell growth.Results. Although leptin was not detected in the bladder tissue specimens, a decreased expression of the leptin receptor was observed in most cancer tissue specimens we analyzed. Furthermore, the forced expression of the leptin receptor in T24 bladder cancer cells prevented them from entering the S phase. Conclusions. Our data demonstrated for the first time that the leptin receptor is aberrantly expressed in bladder cancer tissue and is possibly involved in the carcinogenesis of bladder cancer. (C) 2004 Elsevier Inc.

OBJECTIVE:To compare the clinical and urodynamic characteristics of continent and incontinent women with severe uterovaginal prolapse. STUDY DESIGN:Fifty-eight consecutive women with stage III or IV pelvic organ prolapse between June 1998 and December 2001 were enrolled. Each woman had a urinalysis, pelvic examination and urodynamic study and answered a urinary questionnaire. They were divided into clinically continent (n = 20) and incontinent (n = 38) groups. The clinical symptoms and urodynamic results in the 2 groups were compared statistically with the chi 2 test, Fisher's exact test and Mann-Whitney U test. RESULTS:Incontinent women with severe genital prolapse were more likely to report urinary frequency, urgency and nocturia than were continent women (P < .05). However, the incidence of voiding hesitancy was significantly higher for members of the continent group as compared to the incontinent group (P = .002). With respect to urodynamic variables, including detrusor pressure at peak flow, maximal urethral closure pressure and pressure transmission ratio, significantly higher values occurred in the continent group as compared with the incontinent group; they were 38 (range, 12-66) vs. 24 cm H2O (range, 10-49) (P < .01), 84 (range, 39-117) vs. 63 cm H2O (range, 45-84) (P = .033) and 102% (range, 66-135) vs. 66% (range, 14-98) (P = .019), respectively. All other parameters and the incidence of bladder outlet obstruction and detrusor instability did not differ significantly between the 2 groups (P > .05). CONCLUSION:The results of this study suggest that severe uterovaginal prolapse could produce obstructive symptoms and prevent or reduce urinary leakage, but whether urethral kinking or external urethral compression causes the obstruction remains unclear. More studies on different types of isolated pelvic organ prolapse are needed to elucidate the mechanism, and specific strategies can be developed to aid urogynecologists in their goal of restoring normal anatomy.

Objective. To explore the possibility of using early second trimester amniotic fluid leptin levels as a predictor of pregnancy outcome in twin pregnancy.Study design. Amniotic fluid leptin levels from 18 twin-pregnant women in early second trimester were analyzed for their correlation with gestational age at delivery and fetal birthweight. Leptin levels in 16 amniotic fluid samples collected from small for gestational age (SGA) twin pregnancies were compared with those in 20 amniotic fluid samples collected from non-SGA twin pregnancies.Results. A significant correlation was observed between amniotic fluid leptin levels and gestational age at delivery (r=0.71, p<0.001) as well as fetal birthweight (r=0.72, p<0.001). There was also a significant correlation between gestational age at delivery and fetal birthweight (r = 0.92, p < 0.001). The average gestational age at delivery was 30.4 +/- 1.4 weeks in the SGA group, with a mean birthweight of 1552 +/- 200 g at delivery. For the non-SGA group, the values were 37.3 +/- 0.5 weeks and 2759 +/- 115 g (p < 0.001), respectively. Amniotic fluid leptin levels were found to be significantly igher (p < 0.001) for women in the SGA group (11.4 +/- 1.5 ng/mL) than for those in the non-SGA group (5.4 +/- 10. 5 ng/mL).Conclusion. Higher amniotic fluid leptin levels in early second trimester were associated with both lower gestational age at delivery and lower birthweight. Our results suggest that amniotic fluid leptin levels in early second trimester may be a good marker for the prediction of perinatal complications in twin pregnancy.

OBJECTIVE:Leptin and its receptor are the key players in the regulation of energy balance and body weight control. However, their roles in gynecological malignancies are mostly unclear. In this study, we analyzed the expression and possible involvement of leptin and the leptin receptor in the pathogenesis of endometrial cancer.METHODS:Radioimmunoassay was performed to analyze the serum leptin levels in the endometrial cancer patients, while RT-PCR, immunoblotting, and immunohistochemistry techniques were applied to study the expression of leptin receptor in the endometrioid-type endometrial cancer tissues. Furthermore, BrdU labeling followed by immunofluorescent analysis was used to analyze the effect of leptin receptor overexpression on endometrial cancer cell proliferation.RESULTS:Serum leptin levels are elevated in endometrial cancer patients, but show no significant difference to those of normal controls when normalized by body mass index. On the other hand, lower expression levels of leptin receptor short form (Ob-Ra) were observed in most endometrial cancer tissues, especially in the poorly differentiated ones, and the forced expression of Ob-Ra in RL95-2 endometrial cancer cells prevented them from entering the S-phase.CONCLUSION:In summary, our data demonstrates for the first time that the leptin receptor is aberrantly expressed in endometrial cancer tissues and is possibly involved in the pathogenesis of endometrial cancer.

Background:Adiponectin is an adipocyte-secreted hormone. Serum adiponectin levels are inversely correlated with insulin resistance, and low plasma adiponectin levels have been demonstrated in type 2 diabetic patients. However, information on serum adiponectin levels in gynecological disorders is mostly lacking. In this study, we analyzed the possible correlation between serum adiponectin levels and uterine leiomyomas. Methods: Serum adiponectin levels, determined by radioimmunoassay, were compared in women with uterine leiomyomas (n = 47) and normal controls (n = 46). Results: Serum adiponectin levels in women with uterine leiomyomas (16.4 ± 0.9 µg/ml) were significantly lower (p < 0.05) than those in the normal controls (19.5 ± 1.0 µg/ml). Furthermore, there was an inverse correlation between serum adiponectin levels and body mass index, in both women with leiomyomas and normal controls. Conclusions: Serum adiponectin levels are significantly decreased in myomatous patients.

Objective. To analyze the possible involvement of leptin in uterine leiomyomas.Study design. Serum leptin levels, determined by radioimmunoassay, were compared in myomatic (n = 50) and the normal (n = 50) women.Results. A significant correlation was found between serum leptin levels and body mass index in both the myomatic women (r = 0.76, p < 0.001) and the normal women (r = 0.56, p < 0.001). Serum leptin levels in the myomatic women (9.3 +/- 0.6 ng/mL) were significantly lower ( p < 0.001) than those in the normal women (13.6 +/- 1.2 ng/mL). In addition, the ratios of serum leptin levels/body mass index in the myomatic women (0.38 +/- 0.02) were significantly lower than those in the normal women (0.57 +/- 0.04) ( p < 0.001). A significant correlation was found between the ratios of serum leptin levels/body mass index and body mass index (r = 0.59, p < 0.001) in the normal women, but not in the myomatic women (r = 0.27, p = 0.061).Conclusion. The lower plasma leptin levels observed in the women with myomas were independent of body mass index, and unlike the normal women there was no significant up-regulation of leptin production in response to increased adiposity.

Objective: To assess the effect of laparoscopic hysterectomy (LH) on the mobility and position of bladder neck (BN) and urinary symptoms.Design: We assessed the BN and urinary symptoms of 151 patients by introital ultrasonography and questionnaires before and after LH.Sample: One hundred and fifty-one women who underwent LH from June 1999 to June 2001.Results: A significant decrease was noted in the number of women exhibiting one or more urinary symptoms from 81 (53.6%) preoperatively to 58 (38.4%) postoperatively (P < 0.01). The incidence of urinary frequency, mild stress incontinence and nocturia decreased significantly after laparoscopic hysterectomy (P < 0.01). Changes in other urinary symptoms following hysterectomy showed no statistical significance (P > 0.05). During straining, the postoperative position of the BN localised more dorsally (P < 0.01) and the ventral mobility of the BN decreased significantly following surgery (P < 0.05). There was no significant difference in the location of the BN with respect to the pubis at rest and during straining, in the cephalocaudal direction, before and after hysterectomy (P > 0.05).Conclusion: Some patients experienced a substantial improvement of preoperative urinary symptoms following LH, partly as a result of a decrease in the hypermobility of BN.

Exposure to arsenic has been reported to cause DNA damage and eventually the occurrence of bladder, lung and skin cancers. A previous report has demonstrated that arsenite-induced phosphorylation of Mre11, a protein involved in the repair of DNA double strand breaks (DSBs), is M phase-dependent and requires the Nijmegen breakage syndrome (NBS) protein, NBS1 [DNA Repair 1 (2002) 137]. Furthermore, arsenite treatment arrests cells at the M phase and the cells eventually go through apoptosis [Biochemical Pharmacology 60 (2000) 771]. Here we demonstrate that arsenite treatment enhances the generation of nitric oxide (NO), and that the enhanced NO generation is dominant at the G2/M phase. Arsenite-induced NO generation is impaired in DSB repair-defective NBS cells, but not in NBS1-reconstituted NBS cells, suggesting NBS1 is required for effective NO generation. In summary, our study showed, for the first time, that arsenite-induced NO generation is cell-cycle- and NBS1-dependent.

OBJECTIVES:To study the correlation between amniotic fluid leptin levels and maternal serum leptin levels during the early second trimester, and to determine whether the ratios of amniotic fluid leptin levels to maternal serum leptin levels are elevated in pregnant women who subsequently develop preeclampsia.STUDY DESIGN:Samples from 120 pregnant women were included in this prospective study, of which 20 were from pregnant women who subsequently developed preeclampsia and 100 were from normal pregnant women. Both the amniotic fluid and the maternal serum leptin levels were ascertained by radioimmunoassay (RIA).RESULTS:A strong correlation between amniotic fluid leptin levels and maternal serum leptin levels was observed in both preeclamptic and normal pregnant women. In addition, the ratios of amniotic fluid leptin levels to maternal serum leptin levels were positively correlated to amniotic fluid leptin levels, but negatively correlated to maternal serum leptin levels. Furthermore, the ratios of amniotic fluid leptin levels to maternal serum leptin levels in preeclamptic women were significantly higher than those in normal pregnant women.CONCLUSIONS:Amniotic fluid leptin levels correlated with maternal serum leptin levels during the early second trimester. The ratios of amniotic fluid leptin levels to maternal serum leptin levels were elevated in preeclamptic women. However, the maternal serum leptin levels themselves showed no such elevation. Therefore, this elevated ratio may be a marker at the early stage of pregnancy in preeclamptic women.

Annonaceous acetogenins are a group of potential anti-neoplastic agents isolated from Annonaceae plants. In this study, we purified annonacin, a cytotoxic mono-tetrahydrofuran acetogenin, from the seeds of Annona reticulata and analyzed its biological effects. Herein, we have shown that annonacin caused significant cell death in various cancer cell lines. T24 bladder cancer cells at the S phase were more vulnerable to the cytotoxicity of annonacin. Furthermore, annonacin activated p21 in a p53-independent manner and arrested T24 cells at the G1 phase. It also induced Bax expression, enhanced caspase-3 activity, and caused apoptotic cell death in T24 cells. In summary, these results suggest that annonacin is potentially a promising anti-cancer compound.