Recent studies suggested that the prostate cancer may arise from prostate cancer stem cells that share some same characteristics with normal stem cells. The purpose of this study was to detect the differences of Nanog expression between PC3 prostate cancer cell line and its tumor stem cells, and the relationship was preliminarily examined between Nanog and prostate cancer and its tumor stem cells. By using magnetic active cell sorting (MACS), we isolated a population of CD44(+)/CD133(+) prostate cancer cells that display stem cell characteristics from PC3 cell line. Immunohistochemistry revealed positive expressions of CD44, CD133 and α(2)β(1)-integin in the isolated cells. CCK-8 analysis showed that isolated cells had a strong proliferative ability. The formation of the cell spheres in serum-free medium and holoclones in serum-supplied medium showed that the cells were capable of self-renewing, indicating that the isolated cells were a population of cancer stem-like cells derived from PC3 cell line. Western blotting exhibited that the isolated cells had higher experession of Nanog, an embryonic stem marker, as compared with PC3 cells. Our study showed that Nanog might be helpful in sustaining the self-renewal and the undifferentiation of prostate cancer stem cells, and may serve as a marker for prostate cancer stem cells for isolation and identification.

BACKGROUND: Neuronal apoptosis produced a main effect in secondary injury after injury of central nervous system. Recently, minocycline has been reported to protect neurologic function evidently by decreasing apoptosis.OBJECTIVE:To study the effect of minocycline on apoptosis and expression of related factor, and protective effect of minocycline on neurologic function of injured spine after spinal cord injury (SCI) in rats.DESIGN: A randomized controlled animal study.SETTING: Tongji Hospital Affiliated to Tongji Medical College,Huazhong University of Science and Technology.MATERIALS: The experiment was performed at the Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology between November 2003 and December 2004. A total of 36male SD rats were divided into four groups: ①Blank control group (group A) with 6 rats, ②injured group (group B) with 10 rats, ③minocycline treatment group (group C) with 10 rats, and ④methylprednisolone treatment group (group D) with 10 rats.METHODS: The animal SCI model was established. Spinal cords of rats in the group A were exposed without injury. Spines of rats in the group B were injured without treatment. Rats of group C was treated with minocycline (50 mg/kg) by intraperitoneal injection at 1 hour after injury, and 24 hours later another 50 mg/kg was given, and then another 25 mg/kg was injected for 5 days. Methylprednisolone (100 mg/kg) was intraperitoneally injected 1 hour after injury in the group D. The rats in the group A and the group B were treated with saline, and the method of administration was the same with that of the group C.MAIN OUTCOME MEASURES: ①Neurofunction evaluation of rats in each group. ②Expressions of Bcl-2 family members (Bcl-2, Bcl-xl, and Bax) were tested with immunocytochemical method. ③Apoptosis index of spinal tissue of rats of each group.RESULTS: A total of 36 rats were involved in the result analysis. ①Neurofunction of rats in the group C increased as compared with the group B (P ＜ 0.05), and there was insignificant difference between group C and the group D (P ＞ 0.05). ②Bcl-2 positive expressive cells in the group C was more than that in the group B, and showed obvious difference between the two groups [(62.53±7.76)%, (35.14±3.70)%,P ＜ 0.05]. There was insignificant difference between expressions of Bcl-xl and Bax in group C as compared with the group B (P ＞ 0.05). ③The number of apoptosis positive cells in the group B was more than that in the group C and D [(44.63± 5.90)%, (32.71± 5.26)%, (34.31± 6.84)%,P ＜ 0.05].CONCLUSION: Minocycline can up-regulate the expression of Bcl-2, and raise the ratio of Bcl-2/Bax, so as to inhibit the neural apoptosis of spine,which have protective effect on neurologic function of injured spine.

Recent studies suggested that the prostate cancer may arise from prostate cancer stem cells that share some same characteristics with normal stem cells.The purpose of this study was to detect the differences of Nanog expression between PC3 prostate cancer cell line and its tumor stem cells,and fhe relationship was preliminarily examined between Nanog and prostate cancer and its tumor stem cells.By using magnetic active cell sorting (MACS),we isolated a population of CD44+/CD133+ prostate cancer cells that display stem cell characteristics from PC3 cell line.lmmunohistochemistry revealed positive expressions of CD44,C D 133 and α2β1-integin in the isolated cells.CCK-8 analysis showed that isolated cells had a strong proliferafive ability.The formation of the cell spheres in serum-free medium and holoclones in serum-supplied medium showed that the cells were capable of self-renewing,indicating that the isolated cells were a population of cancer stem-like cells derived from PC3 cell line.Western blotting exhibited that the isolated cells had higher experession of Nanog,an embryonic stem marker,as compared with PC3 cells.Our study showed that Nanog might be helpful in sustaining the self-renewal and fhe undifferentiation of prostate cancer stem cells,and may serve as a marker for prostate cancer stem cells for isolation and identification.

Aim To investigate the effect of CDR132L(miR-132 antisense oligonucleotide)on vascular remode-ling and function in mice with hypertension and hyperlipidemia,and explore its possible mechanism.Methods A total of 30 8-week-old male C57BL/6 mice were randomly divided into three groups:control group,model group and CDR132L group,with 10 mice in each group.The control group received with a standard diet while the model group and CDR132L group received N-nitro-L-arginine methyl ester(L-NAME)and high-fat diet to induce hypertension and hyperlip-idemia.The CDR132L group was administered with intraperitoneal injection of CDR132L at a dose of 20 mg/kg once weekly for six consecutive weeks,whereas the control group and the model group were given intraperitoneal injection of an equivalent volume of normal saline.The tail-cuff method was utilized for blood pressure measurement,blood lipid and glucose levels were assayed by an automatic biochemical analyzer,the thoracic aorta structure was observed by HE staining,endothelium-dependent relaxation of the thoracic aorta was evaluated by the vascular ring test,the expression level of miR-132 in the thoracic aorta was measured by qPCR,the protein expression levels of Gab1 and endothelial nitric oxide synthase(eNOS)in the thoracic aorta were determined by Western blot.Results Compared with the control group,the model group demonstrated notable rises in systolic and diastolic blood pressure,serum triglyceride,total cholesterol lev-els,and body weight.Moreover,the intima of thoracic aorta and the thickness of vascular wall was uneven,the smooth muscle cells of the tunica media were arranged irregularly,with a large amount of fat deposition in the vascular wall,and the endothelium-dependent relaxation response of thoracic aorta was decreased(P＜0.05).The expression level of miR-132 in the thoracic aorta was significantly increased(P＜0.05),while the expression level of Gabl and eNOS protein was markedly decreased(P＜0.05).Compared with the model group,the CDR132L group showed no significant differences in systolic and diastolic blood pressure,serum triglyceride and total cholesterol levels,as well as body weight(P＞0.05).However,the CDR132L group exhibited a complete and smooth intima of the thoracic aorta with minimal intravascular lipid deposition,the thickness of the vascular wall was uniform,the smooth muscle cells of the tunica media were arranged order-ly,accompanied by enhanced endothelium-dependent relaxation response of the thoracic aorta(P＜0.05).The expression level of miR-132 in the thoracic aorta was significantly decreased(P＜0.05),while the expression levels of Gab1 and eNOS protein were significantly increased(P＜0.05).Conclusion CDR132L can improve vascular remodeling and endothelium-dependent relaxation in hypertensive and hyperlipidemia mice,which may be related to the decrease of miR-132 expression level and the up-regulation of Gab1 and eNOS protein expression levels in the thoracic aorta.