Objective To observe after the NIBP (NIK and IKK beta binding protein) gene transfected into colon cancer cell lines HT29, the migration of cells and expression of p65, MMP2, MMP9 mRNA and proteins. Methods The experimental group: divided into without transfection HT29 cell (HT29 group) and transfection no-load HT29 cell (HT29-NC group) and transfection NIBP HT29 cell (HT29-NIBP steady group). Using the Transwell test to detect cell migration ability. Q-PCR method to detect the mRNA expressions of NIBP , p65, MMP2, MMP9. Western Blot method to detect the expressions of NIBP, p65, phosphorylation p65 (p-p65) proteins. The method of ELISA was used to detect the secretion of matrix metalloproteinase (MMP)-2, MMP-9. Results The high expression of NIBP may enhance the migration ability of colon cancer cell lines HT29 , increasing the expression of p-p65, MMP2, MMP9 mRNA and proteins (P < 0.05). Conclusion NIBP potently enhances colon cancer HT29 cell migration and invasion. Activation of NF-κB signaling pathways resulted in up-regulation of MMP-2 and MMP-9 maybe one of its molecular mechanisms.

[ ABSTRACT] AIM:To investigate the effects of sphingosine kinase l ( SphK1) and focal adhesion kinase ( FAK) on the epithelial-mesenchymal transition ( EMT) of human colon cancer HCT 116 cells.METHODS:Human colon cancer HCT116 cells were divided into 3 groups.N, N-dimethylsphingosine (DMS) was used to suppress the activity of SphK1. PF573228 was used to suppress the activation of FAK .The cells treated with equal volume of culture medium severed as control group.The cell viability was measured by MTT assay .The protein expression of SphK1, FAK and the EMT relative protein E-cadherin, N-cadherin, vimentin and matrix metalloproteinase (MMP) 2 was analyzed by Western blot.The mR-NA expression of SphK1, sphingosine-1-phosphate (S1P), FAK, E-cadherin and vimentin was detected by real-time PCR. The ability of tumor cell migration was measured by wound-healing assay.RESULTS:The cell viability of HCT116 cells was suppressed by DMS and PF 573228 in dose and time dependent manners .DMS significantly suppressed the expression of SphK1, FAK, N-cadherin, vimentin and MMP2, meanwhile enhanced the expression of E-cadherin.PF573228 reduced the expression of FAK , SphK1, N-cadherin, vimentin and MMP2, meanwhile increased the expression of E-cadherin (P<0.01).In addition, the migration ability of HCT116 cells was significantly decreased by treating with DMS and PF573228 (P<0.01).Compared with control group , the mRNA expression of FAK, SphK1, S1P and vimentin was de-creased, while the expression of E-cadherin was increased significantly in PF573228 group and DMS group (P<0.05). CONCLUSION:SphK1 and FAK signaling pathways may play an important role in the occurrence of EMT in the colon cancer HCT116 cells.

Objective To investigate the expression of DRAM in breast cancer cell line MCF-7 in radiation-induced autophagy. Methods GFP-LC3 transfection method was used to observe autophagy bubble.Real time-PCR was used to detect DRAM and MAPLC3 from transcriptional and translational level,respectively. The silencing vector from gene engineering was introduced by calcium phosphate transfection.Results Compared with the control group,GFP-LC3 increased significantly after 8 Gy irradiation by immunofluorescent assay,and the level of MAP-LC3 expression was higher than control group after 8 Gy irradiation by Western blot ( F =5.38,8.72,10.63,15.23,20.78 and 55.23,P ＜ 0.05 ).DRAM protein expression increased significantly at 2 h in the 8 Gy time-dose study,up to maximum at the 32 h( F =116.34,P ＜ 0.05 ).In DRAM model,the expression of LC3 and DRAM decreased significantly (F =20.36 and 40.35,P ＜ 0.05 ) and DRAM expression increased 8 Gy post-irradiation,but still lower than that in 8 Gy irradiation wild-type group.The LC3 expression also decreasaed 8 Gy post-irradiation(F =50.34,P ＜ 0.05 ).Conclusions DRAM plays an important role in irradiation-induced autophagy in breast cancer cell.DRAM might participate in the process and serve as a theoretical target for clinical treatment of breast cancer.

Cemento-ossifying fibroma is a rare benign tumor from periodontium, which usually occurs in mandible body and mandible ramus. It consists of collagen fibrils, fibroblast, and cementoblast. This article reported a case of giant cemento-ossifying fibroma and discussed the clinical features and treatment.
Dental Cementum ; Fibroblasts ; Fibroma, Ossifying ; Humans ; Mandible ; Periodontal Ligament

OBJECTIVETo investigate the alkaloids from Aconitum handelianum.

METHODThe column chromatographic methods were employed for the isolation and purification of the chemical constituents. The structures were elucidated by spectroscopic methods.

RESULTEight diterpenoid alkaloids were isolated and identified as acoforine (1), acoforestinine (2), 14-O-acetylsachaconitine (3), vilmorrianine C (4), vilmorrianine D (5), talatizamine (6), chasmanine (7) and yunaconitine (8).

CONCLUSIONAll compounds were isolated from this plant for the first time.

Aconitum ; chemistry ; Alkaloids ; analysis ; isolation & purification ; Diterpenes ; analysis ; isolation & purification ; Drugs, Chinese Herbal ; analysis ; isolation & purification

Captopril can have nephrotoxic effects, which are largely attributed to accumulated renin and "escaped" angiotensin II (Ang II). Here we test whether angiotensin converting enzyme-1 (ACE1) inhibition damages kidneys via alteration of renal afferent arteriolar responses to Ang II and inflammatory signaling. C57Bl/6 mice were given vehicle or captopril (60 mg/kg per day) for four weeks. Hypertension was obtained by minipump supplying Ang II (400 ng/kg per min) during the second 2 weeks. We assessed kidney histology by periodic acid-Schiff (PAS) and Masson staining, glomerular filtration rate (GFR) by FITC-labeled inulin clearance, and responses to Ang II assessed in afferent arterioles in vitro. Moreover, arteriolar H₂O₂ and catalase, plasma renin were assayed by commercial kits, and mRNAs of renin receptor, transforming growth factor-β (TGF-β) and cyclooxygenase-2 (COX-2) in the renal cortex, mRNAs of angiotensin receptor-1 (AT₁R) and AT₂R in the preglomerular arterioles were detected by RT-qPCR. The results showed that, compared to vehicle, mice given captopril showed lowered blood pressure, reduced GFR, increased plasma renin, renal interstitial fibrosis and tubular epithelial vacuolar degeneration, increased expression of mRNAs of renal TGF-β and COX-2, decreased production of H₂O₂ and increased catalase activity in preglomerular arterioles and enhanced afferent arteriolar Ang II contractions. The latter were blunted by incubation with H₂O₂. The mRNAs of renal microvascular AT₁R and AT₂R remained unaffected by captopril. Ang II-infused mice showed increased blood pressure and reduced afferent arteriolar Ang II responses. Administration of captopril to the Ang II-infused mice normalized blood pressure, but not arteriolar Ang II responses. We conclude that inhibition of ACE1 enhances renal microvascular reactivity to Ang II and may enhance important inflammatory pathways.
Angiotensin II/pharmacology* ; Animals ; Arterioles/metabolism* ; Captopril/pharmacology* ; Hydrogen Peroxide/pharmacology* ; Kidney ; Mice