Objective; To investigate the effect of NDGA, the lipoxygenase inhibitor, on colon cancer cell line HT-29 in vitro from the aspects of cell growth inhibition, cell apoptosis and its impact on the expression of telomerase. Methods: We applied respectively i) MTT to draw the growth curve. ii) inverted phase contrast microscope to observe morphologic change of cells, iii) scanning electron microscope to observe changes of cell's ultra-microstructure and apoptotic body. iv) RT-PCR to detect the changes of the expression of human telomerase reverse transeriptase (hTERT). Results: Different concentrations of NDGA were used to dispose cancer cells separately, with the rise of the drugis concentration, the form of cells became round, the volume waned, cells abscised from the inner surface of the bottle. and growth inhibition became increasing abvious. Also through scaming electron microscope, apoptic bodies could be found colon cancer cell line HT-29 showed positive expression of hTERTmRNA, which became weaker following the rising of the drugs concentration. Conclusions: NDGA which, displays the relying effect of doses, can inhibit the growth of colon cancer cell line HT-29 and induce its apoptosis, telomerase plays an active role in this course.

Objective:To investigate the regulatory effects of sodium butyrate on p53 target genes(p21waf1,bax,and gadd45)in HT-29 colorectal cancer cells and the related mechanisms.Methods:HT-29 cells were cultured in the absence or presence of sodium butyrate.The cell proliferation and cell cycle were studied by MTT and FCM,respectively.Apoptosis was assessed by observing cell morphology,percentage of sub-G_ 1 cells and AnnexinV-FITC.The effects of sodium butyrate on transcription of p21waf1,bax and gadd45 were analyzed by RT-PCR and Western blot.Results:Sodium butyrate inhibited proliferation and induced apoptosis of HT-29 cells in a time-and dose-dependent manner,and it blocked HT-29 cell at G_ 1 phase.Sodium butyrate stimulated p21waf1 and bax expression both at mRNA and protein level in HT-29 cells,but had little effect on the transcription of gadd45.Conclusion:Sodium butyrate can inhibit proliferation and induce apoptosis of HT-29 cells,which might be through up-regulating p21waf1 and bax expression both at mRNA and protein levels.

Objective To study the effect of imbalance of proliferation and apoptosis in the development of colorectal carcinoma(CRC),and the molecular mechanism of the dynamic change.Methods ORC was(induced) with dimethylhydrazine(DMH) in male mice of Kimming strain.The mice were killed in batches in the 12th,18th and 24th weeks of carcinoma induction.The distribution and extent of proliferation and(apoptosis) of the colorectal mucosa,at various intervals,were dynamically observed.Three genes,p21waf1,Bax and Gadd45 were analyzed by RT-PCR,immunohistochemistry and Western blot.Results During the course of carcinoma induction,the mucosas of the model mice showed sequential changes of atypital(hyperplasia),adenoma,and carcinoma.Compared with control group,the PCNA expression of the model group mice was significantly higher(P

AIM: To investigate whether cirrhosis affects the expression of hepatic glucuronosyltransferase (UGT) isoforms and its clinical significance. METHODS: Reverse transcription and polymerase chain reaction (RT-PCR) and Southern hybridization were used to determine the mRNA levels of five UGT isoforms in hepatic tissues. RESULTS: Compared to control group, UGT2B15 and UGT1A6 mRNA level in cirrhosis group increased by 227% and 166%, respectively. Compared to 0 fibrotic grade group, UGT2B15 mRNA levels of 1, 2, 4 fibrotic grade patients were 172%, 208% and 243%, respectively. UGT2B7 mRNA levels of 1, 2, 3 fibrotic grade patients were 156%, 208% and 192%, respectively. Inflammation grade showed no obvious effect on mRNA expression of most UGT isoforms. CONCLUSION: Cirrhosis affectes individual UGT isoform mRNA level. Degree of fibrosis has a significant effect on UGT mRNA expression.

AIM: To evaluate the growth-inhibitory effects of NS-398, a selective cyclooxygenase-2 inhibitor, in human colon cancer HT-29 cells and its possible mechanisms. METHODS: MTT assay was applied to detect the cell proliferation. Flow cytometry was performed to detect apoptosis rate and cell cycle. RT-PCR was used to detect the expression of bcl-2 mRNA and bax mRNA. Alteration of cytoskeleton component F-actin was observed by confocal laser scanning microscope. RESULTS: NS-398 could inhibit growth of HT-29 cells in dose-and time-dependent manners. Flow cytometry revealed that NS-398 could induce apoptosis and cause G0/G1 arrest of HT-29 cells in a dose-dependent manner. After 72 h incubation with NS-398 at different concentrations, the expression level of bcl-2 mRNA was lowered and the ratio of bcl-2 to bax was decreased in HT-29 cells. F-actin was mainly distributed around nuclei forming annular structure in HT-29 cells. After exposure to NS-398, the annular structure around nuclei disappeared and fluorescence intensity of F-actin decreased obviously. CONCLUSION: NS-398 can inhibit the growth effectively and induce apoptosis in HT-29 cells in vitro, which is associated with the down-regulation of bcl-2 to bax ratio and the disruption of cytoskeleton.

AIM: To study the anti-invasive effect of NS-398 on colon cancer cell line HT-29 in vitro an its regulation by CD44v6 and nm23-H1 genes. METHODS: Flow cytometry was used to detect the expression of COX-2 and CD44v6 in HT-29 cells. MTT was used for cell survival rate tests. The modified Boyden chamber model was used for quantitative invasion assay. RT-PCR was used to detect the expression of nm23-H1 mRNA. RESULTS: Flow cytometry analysis showed that COX-2 was positive in HT-29 cells. NS-398 had significant inhibitory effects on invasion of HT-29 cells, which had no relation with its cytotoxicity. NS-398 down-regulated the expression of CD44v6 and up-regulated the expression of nm23-H1 mRNA. CONCLUSION: NS-398 has an anti-invasive effect on HT-29 cells in vitro. Down-regulation of CD44v6 and up-regulation of nm23-H1 may be its underlying mechanisms.

Fas-associated death domain (FADD) is a signal connection protein in Fas/FasL apoptotic path which might play a key role on apoptosis by transferring apoptotic signal. To reveal the intracellular signal transduction molecules involved in the procedure of follicular development in bovine ovary, we cloned FADD gene in bovine ovary tissue with RT-PCR, deleted the termination codon in its cDNA and directionally cloned the amplified FADD gene into eukaryotic expression vector pAcGFP-N1 including AcGFP, successfully constructed the fusion protein recombinant plasmid. After identifying by restrictive enzyme Bgl II/EcoR I and sequencing, transfected pAcGFP-bFADD into CHO-K1 cell mediated by Lipofectamine 2000, observed the expression of AcGFP and detected the transcription and expression of FADD by RT-PCR and Western blotting. The results showed that the cattle FADD was successfully cloned, the pAcGFP-bFADD fusion protein recombinant plasmid was successfully constructed by introducing Bgl II, EcoR I cloning site at two ends of FADD open reading frame and inserting a Kozak sequence before start codon. AcGFP expression was detected as early as 24 h after transfection. The percentage of AcGFP positive cells reached about 65% after 24 h. A 654 bp transcription was amplified by RT-PCR, and 51.4 kD target protein was detected by Western blotting. Construction of pAcGFP-bFADD recombinant plasmid should be helpful for further understanding the mechanism of regulation of FADD on bovine oocytes formation and development.
Animals ; Base Sequence ; CHO Cells ; Cattle ; Cloning, Molecular ; Cricetinae ; Cricetulus ; DNA, Complementary ; genetics ; Fas-Associated Death Domain Protein ; biosynthesis ; genetics ; Female ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Molecular Sequence Data ; Oocytes ; cytology ; Open Reading Frames ; Ovary ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection

Objective To evaluate efficacy and safety of Atorvastatin combined with trimetazidine Sibutramine treating angina pectoris of coronary heart disease.Methods Of computer retrieval China National Knowledge Infrastructure (CNKI),Wanfang database,VIP database and search atorvastatin statins combined with trimetazidine trimetazidine in the treatment of angina pectoris of coronary heart disease:a randomized controlled trial,according to Jadad scale to evaluate the quality of the included studies and extracted data.Meta analysis was performed using RevMan 5.3.Results In accordance with the inclusion criteria were included in the 18 study,a total of 1 848 patients.The software of RevMan 5.3 on cardiovascular events incidence,clinical curative effect,angina pectoris,blood lipid [total cholesterol (TC),triglyceride (TG),low density lipoprotein cholesterol (LDL-C),high density lipoprotein cholesterol (HDL-C)],the improvement of cardiac function [including cardiac index left ventricular end diastolic diameter (LVEDD),left ventricular end systolic diameter (LVESD)] and the adverse reactions of meta analysis showed that compared with pure atorvastatin,atorvastatin combined with trimetazidine can reduce the incidence of cardiovascular events [OR =0.19,95％ Cl (0.11,0.35),P＜0.01],reduce seizure frequency and duration of angina [WMD =-1.52,95％ CI (-1.84,-0.99),P ＜ 0.01;WMD =-1.80,95％ CI (-2.20,-1.50),P ＜0.01],improve the clinical efficiency of [OR =4.78,95％ Cl (3.54,6.47),P ＜ 0.01] display.Blood lipid and cardiac ultrasound index show that atorvastatin combined with trimetazidine can better improve the patient's LVEDD [WMD =-2.69,95％ CI (-4.39,-0.98),P ＜ 0.01] LVESD [WMD =-6.92,95 ％ CI(-11.82,-2.02),P ＜ 0.01].The decrease of serum level of TC,TG,LDL-C was better than atorvastatin monotherapy,but there were no statistically significant differences in the improvement level of serum HDL-C in the included patients (P =O.17).In the included studies,atorvastatin combined with trimetazidine can effectively reduce the incidence of adverse reactions in patients [0R=0.33,95％ CI (0.13,0.85),P=0.02].Conclusions Atorvastatin combined with trimetazidine is safer and more effective than atorvastatin alone in the treatment of coronary heart disease angina pectoris.