Castration impairs erectile organ structure and function by inhibiting autophagy and promoting apoptosis of corpus cavernosum smooth muscle cells in rats.

OBJECTIVE: The aim of this study was to determine the changes and underlying mechanisms of erectile organ structure and function in castrated rats. In addition, the regulatory effects of an androgen on autophagy and apoptosis in corpus cavernosum smooth muscle cells (CCSMCs), especially the regulatory effect of androgen on the BECN 1-Bcl-2 interaction, were investigated. METHODS: Male Sprague-Dawley rats were divided into three groups (30/group): control group, castration group, and castration with testosterone supplementation group. The erectile function was examined both in vivo and in vitro, by electric stimulation of the cavernous nerve and corpus cavernosum strip bath test, respectively. Transmission electron microscopy, TUNEL assay, Masson's trichrome staining, immunohistochemistry, and western blotting were performed to determine the levels of autophagy and apoptosis, and the structural changes in corpus cavernosum. RESULTS: Compared with control group, the castration group showed (1) lower erectile function: lower intracavernosal pressure/mean arterial pressure ratio, lower systolic and diastolic capability of corporal strips, and reduced expressions of eNOS and nNOS; (2) greater fibrosis: decreased smooth muscle/collagen ratio, lower expression of α-SMA, and higher expression of TGF-ß1; (3) inhibited autophagy: decreased autophagosomes, lower expressions of BECN1 and LC3-II; and (4) enhanced apoptosis: higher apoptotic index and decreased Bcl-2/Bax ratio. Testosterone supplementation partially improved the effects of castration. CONCLUSIONS: Castration attenuates erectile function and induces corporeal fibrosis by inhibiting autophagy and promoting apoptosis of CCSMCs in rats. Therefore, our study highlights the important role of androgens in maintaining the integrity of the structure and function of corpus cavernosum in rats through counter-regulation of autophagy and apoptosis, mainly by regulating BECN 1-Bcl-2 interaction.

Asparagus polysaccharide and gum with hepatic artery embolization induces tumor growth and inhibits angiogenesis in an orthotopic hepatocellular carcinoma model.

Liver cancer is one of leading digestive malignancies with high morbidity and mortality. There is an urgent need for the development of novel therapies for this deadly disease. It has been proven that asparagus polysaccharide, one of the most active derivates from the traditional medicine asparagus, possesses notable antitumor properties. However, little is known about the efficacy of asparagus polysaccharide as an adjuvant for liver cancer chemotherapy. Herein, we reported that asparagus polysaccharide and its embolic agent form, asparagus gum, significantly inhibited liver tumor growth with transcatheter arterial chemoembolization (TACE) therapy in an orthotopic hepatocellular carcinoma (HCC) tumor model, while significantly inhibiting angiogenesis and promoting tumor cell apoptosis. Moreover, asparagine gelatinous possessed immunomodulatory functions and showed little toxicity to the host. These results highlight the chemotherapeutic potential of asparagus polysaccharide and warrant a future focus on development as novel chemotherapeutic agent for liver cancer TACE therapy.

KAT5 and KAT6B are in positive regulation on cell proliferation of prostate cancer through PI3K-AKT signaling.

Histone modifications play important roles in the tumorigenesis and progression of prostate cancer (PCa) and genes involved in histone modifications are seemed as ideal targets for treatment of PCa patients. However, clinical trials have shown that those existing drugs exert the minimal antitumor activity and excess adverse effects on PCa patients. Therefore, it is of great interest to figure out novel specific biomarkers to guide the development of new drugs. In present study, an RNAi screening with 44 genes involved in histone modifications was applied to a PCa cell line, Du145. The results showed that nine genes were in positive regulation of Du145 cell growth. Then four selected genes (KAT2B, KAT5, KAT6B and HDAC1) were found to exert this effect by a gene-specific manner when silenced. And then KAT5 or KAT6B silenced cells were subjected to DNA microarray analysis. The common differentially expressed genes were analyzed by Ingenuity Pathway Analysis (IPA) and found that PDEF signaling, EIF2 signaling and PI3K signaling was suppressed following by KAT5 or KAT6B silencing. Subsequent immunoblotting assay showed that AKT signaling was inhibited, which suggested that KAT5 or KAT6B regulates cancer cell growth through PI3K-AKT signaling. Together with our published data [31] that AURKA inhibitoin increased drug sensitivity of DU145, our work demonstrated the underlying mechanism that how the acetylation enzyme regulates cancer cells survial and might provide potential therapeutic targets for prostate cancer patients in future epigenetic drug development.

AURKA suppression induces DU145 apoptosis and sensitizes DU145 to docetaxel treatment.

The palliative therapy effect by docetaxel for CRPC patients makes it urgent to improve the therapy. It was suggested that PI3K and androgen receptor-directed combination therapy may be effective for prostate cancer (PCa) patients PTEN negative. However, for those patients PTEN positive, the mechanism of anti-apoptosis survival of cancer cells is not yet well defined. Amplification of AURKA has been detected in 5% of PCa. In this work, Du145, a PTEN positive PCa cell model, was employed to investigate the role of aurora kinase a (AURKA) on cell growth. Inhibition of AURKA expression by shRNA markedly reduced prostate cancer cell viability. Furthermore, we demonstrate that AURKA inhibition induced a remarkable downregulation of AKT activity and Bax induction. Moreover, specific inhibition of the activity of AURKA, but not other aurora family members, by small molecular chemical inhibitors induced significant cell killing effects. Notably, AURKA inhibition sensitized prostate cancer cells to docetaxel treatment. Our work suggests that AURKA-directed monotherapy or combination therapy with docetaxel could be a potent treatment for PCa patients in future.

Effects of oral finasteride on erectile function in a rat model.

INTRODUCTION: Many clinical studies reported finasteride-related erectile dysfunction, but to date, few animal experiments have focused on it. AIM: To investigate the effects of oral finasteride on erectile function in a rat model. MAIN OUTCOME MEASURES: Erectile responses and morphological changes. METHODS: Adult, male Sprague-Dawley rats were divided into four groups (25/group): (i) control; (ii) castration; (iii) castration with testosterone (T) replacement; and (iv) oral finasteride treatment. Four weeks later, erectile function was measured by the ratio of intracavernosal pressure and mean arterial blood pressure upon electrical stimulation of the cavernous nerve. Serum T and dihydrotestosterone (DHT) and intraprostatic DHT were measured. The weights and histopathological features of the penile corpus cavernosum and prostate were examined. RESULTS: Serum T and DHT and intraprostatic DHT concentrations, erectile function, and mean weights of the corpus cavernosum and prostate were lowest in group 2. There was no significant difference in the serum T concentration and erectile function between groups 4 and 1. However, the serum and intraprostatic DHT concentrations were significantly lower in group 4 than in group 1 (both P < 0.001). The tissue weights of the corpus cavernosum and prostate were reduced by 25.9% and 92.3% in group 4 compared with group 1 (both P < 0.001). Histopathology revealed a significant atrophy of the prostate in groups 2 and 4. There was a significant decrease in the smooth muscle content in group 2, but not in groups 3 and 4. CONCLUSIONS: In a rat model, finasteride treatment for 4 weeks reduces the weight of the corpus cavernosum but appears not to affect the erectile responses to electrical stimulation of the cavernous nerve. As erection is a complex process involving important signaling in the brain, further studies are necessary to demonstrate the long-term effects of finasteride on both central and peripheral neural pathways of erection.

Engagement of integrinß1 induces resistance of bladder cancer cells to mitomycin-C.

OBJECTIVE: To identify whether integrinß1 subunit is responsible for the resistance of bladder cancer cell to the therapeutic drug mitomycin-C (MMC), when grown on fibronectin (FN). MATERIALS AND METHODS: The expression of integrinß1 on bladder cancer T24 and 5637 cells was examined by the flow cytometer. The adhesion of cells to plates with the absence or presence of FN was determined. Analysis of apoptosis induced by MMC was assessed using the flow cytometer in combination with an integrinß1-blocking antibody or siRNA targeting the coding region of integrinß1. Western blot was used to study the expression change of integrinß1 and its downstream molecules. RESULTS: Bladder cancer T24 and 5637 cells express high level of integrinß1 (87.3% ± 2.3 and 90.1% ± 1.9, respectively). Cellular adhesion to FN was significantly reduced by the blocking of integrinß1. Blocking or silencing of integrinß1 significantly abolished the drug resistance of cells grown on FN to MMC (P <.05) and inhibited the activation of survival signals phosphoinositide-3 kinase (PI3-K)/Akt. CONCLUSION: Integrinß1-mediated cellular adhesion to FN confers drug resistance to MMC on bladder cancer cells. Knockdown of integrinß1 may abolish the drug resistance phenotype and sensitize bladder cancer cells to MMC.

Vasoactive intestinal polypeptide, an erectile neurotransmitter, improves erectile function more significantly in castrated rats than in normal rats.

OBJECTIVE: • To investigate the regulatory role of androgen in VIP-mediated erectile effect. Androgen is essential for physiological erection. Vasoactive intestinal polypeptide (VIP) is an important erectile neurotransmitter. While previous studies demonstrated that VIP expression in the penis was androgen-independent, it remains controversial whether androgen has any effect on VIP-mediated erection. MATERIALS AND METHODS: • Male SD rats were divided into a control group, a castration group, and a castration-with-testosterone-replacement group. Four weeks later, each group was subdivided into low and high-dose VIP subgroups and subjected to intracavernous injection of 0.5 and 2 µg VIP, respectively. • Erectile function was tested by recording intracavernosal pressure (ICP) and mean arterial blood pressure (MAP) before and after VIP injection. • The expressions of the VIP-receptor (VPAC2), G-protein stimulatory and inhibitory alpha subunits (Gs-α, Gi-α), and PDE3A in rat corpus cavernosum (CC) was qualified by real-time PCR and Western blot analysis. RESULTS: • Castration reduced erectile function while testosterone restored it. VIP improved erectile function in a dose-dependent manner. • High-dose VIP significantly enhanced erectile function in castrated rats and there was no difference of ICP/MAP among three groups after injection of high-dose VIP. • Low-dose VIP also resulted in a higher improvement of erectile function in castrated rats, although the ICP/MAP was lower in these rats than in the other two groups. VPAC2 and Gs-α were up-regulated while Gi-α and PDE3A were down-regulated in CC of castrated rats. CONCLUSION: • VIP improves erectile function much more significantly in hypogonadal condition, mainly due to the higher expression of VPAC2, Gs-α, and lower expression of Gi-α and PDE3A in CC of castrated rats. Androgen may negatively regulate the erectile effect of VIP.

Relationship between CT grouping and complications of liver cirrhosis.

BACKGROUND: Imaging examination is important for hepatic cirrhosis. But the relationship between magnetic resonance (MR), computed tomography (CT) or ultrasound findings and pathological groups, degree, or reserve function of the cirrhotic liver is not clear. In this study, we investigated the relationship between the CT groupings of liver cirrhosis and its complications and clinical conditions. METHODS: The CT findings in 357 patients with liver cirrhosis were grouped. The complications were analyzed, included splenomegaly, varicose collateral veins, ascites, pleurorrhea, primary liver carcinoma, gallbladder stone, etc. Blood routine (BRt), and serum usea nitrogen (SUN), creatinine and uric acid were measured and hypersplenia and liver-kidney syndrome were estimated. RESULTS: Three hundred and fifty-seven patients with cirrhosis were divided into homogeneous group (87 patients, 24.4%), segmental group (41, 11.5%), and nodal group (229, 64.2%). The grade of spleen enlargement in the segmental and the nodal groups was significantly greater than that in the homogeneous group (P<0.01 and P<0.001). The patients with varices were shown in a descending order in the segmental group (70.7%), the nodal group (17.0%) and the homogeneous group (2.3%), respectively. Significant difference was observed among the 3 groups (P<0.001). Ascites was seen in 182 patients (79.5%) of the nodal group, in 11 patients (26.8%) of the segmental group and in 9 patients (10.3%) of the homogeneous group (P<0.001). Sixty-eight patients (29.7%) in the nodal group had primary liver carcinoma and 1 (2.4%) in the segmental group and 5 (5.8%) in the homogeneous group (P<0.001). The number of patients with decreased concentration of hematoglobin in the nodal group was more than that in the homogeneous group (P<0.001). The mean values of hematoglobin and platelet in the nodal group and the segmental group were significantly lower than those in the homogeneous group (P<0.05). The number of patients with increased concentration of SUN in the nodal group and the segmental group was more than that in the homogeneous group (P<0.005). The concentration of SUN in the nodal group was significantly higher than that in the homogeneous group (P<0.002). CONCLUSION: There is a close relationship between the grouping of liver cirrhosis by CT findings and complications caused by the cirrhosis. The grouping is significant for estimating clinical conditions.

[A study on relationship between CT grouping and complication of liver cirrhosis].

OBJECTIVES: To probe into the relationship between the CT grouping of liver cirrhosis and its complications as well as clinical condition. METHODS: On the basis of CT findings in 357 cases with liver cirrhosis, CT grouping of the cirrhosis was performed. The complications of the cirrhosis, including enlargement of spleen, varicose collateral veins, ascites, pleurorrhea, primary liver carcinoma, gallbladder stone, etc, of all groups were analyzed. According to examination the blood routine, and the serum urea nitrogen (SUN), creatinine and uric acid, the condition of spleen function accentuation and liver-kidney syndrome were estimated. RESULTS: Three hundred and fifty-seven cases with cirrhosis were divided into homogeneous group (87 cases, 24.4%), segmental group (41 cases, 11.5%) and nodal group (229 cases, 64.1%). The grade of spleen enlargement in the segmental group and the nodal group was significantly greater than that in homogeneous group. The cases with varicose in the segmental group was the most (70.7%), in the nodal group next (17.0%) and in the homogeneous group the least (2.3%). There was significant difference among three groups. In the nodal group, there was ascites in 182 cases (79.5%) and significantly more than that in the segmental group (11 cases, 26.8%) and the homogeneous group (9 cases, 10.3%), and the former significantly more than the latter. There were 68 cases (29.7%) with primary liver carcinoma in the nodal group, and significantly more than that in the segmental group (1 case, 2.4%) and the homogeneous group (5 cases, 5.7%). The cases with hemoglobin decrease in the nodal group were significantly more than that in the homogeneous group. The averages of hemoglobin and platelet in the nodal group and the segmental group were significantly lower than that in the homogeneous group. The cases with SUN increase in the nodal group and the segmental group were significantly more than that in the homogeneous group. The concentration of SUN in the nodal group was significantly higher than that in the homogeneous group. CONCLUSIONS: There are close relationship between the grouping of liver cirrhosis on basis of CT findings and complications of the cirrhosis. The practice of grouping might be useful for estimating clinical condition